Sample records for boat basic oligonucleotide

  1. Clay catalysis of oligonucleotide formation: kinetics of the reaction of the 5'-phosphorimidazolides of nucleotides with the non-basic heterocycles uracil and hypoxanthine (United States)

    Kawamura, K.; Ferris, J. P.


    The montmorillonite clay catalyzed condensation of activated monocleotides to oligomers of RNA is a possible first step in the formation of the proposed RNA world. The rate constants for the condensation of the phosphorimidazolide of adenosine were measured previously and these studies have been extended to the phosphorimidazolides of inosine and uridine in the present work to determine of substitution of neutral heterocycles for the basic adenine ring changes the reaction rate or regioselectivity. The oligomerization reactions of the 5'-phosphoromidazolides of uridine (ImpU) and inosine (ImpI) on montmorillonite yield oligo(U)s and oligo(I)s as long as heptamers. The rate constants for oligonucleotide formation were determined by measuring the rates of formation of the oligomers by HPLC. Both the apparent rate constants in the reaction mixture and the rate constants on the clay surface were calculated using the partition coefficients of the oligomers between the aqueous and clay phases. The rate constants for trimer formation are much greater than those dimer synthesis but there was little difference in the rate constants for the formation of trimers and higher oligomers. The overall rates of oligomerization of the phosphorimidazolides of purine and pyrimidine nucleosides in the presence of montmorillonite clay are the same suggesting that RNA formed on the primitive Earth could have contained a variety of heterocyclic bases. The rate constants for oligomerization of pyrimidine nucleotides on the clay surface are significantly higher than those of purine nucleotides since the pyrimidine nucleotides bind less strongly to the clay than do the purine nucleotides. The differences in the binding is probably due to Van der Waals interactions between the purine bases and the clay surface. Differences in the basicity of the heterocyclic ring in the nucleotide have little effect on the oligomerization process.

  2. Guam Boat-based Creel Survey (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Similar to other boat-based survey in basic design, this system is run by the Div. of Aquatic and Wildlife Resources (DAWR) and has been in operation since about...

  3. New Russian Combat Small Boats

    Directory of Open Access Journals (Sweden)

    Aleksandr F. Mitrofanov


    Full Text Available The article presents an overview of small combat boats. The author provides a description and gives an analysis of the characteristics of the boat "Raptor", boat "BK-16", boat "Strizh-4-1 DSh", and assault boat "BK-10".

  4. 77 FR 31147 - National Safe Boating Week, 2012 (United States)


    ... waterways. By planning ahead and taking basic safety precautions, boat operators and passengers can help... organizations and governments across our country to prevent loss of life, personal harm, and property damage... recognition of the importance of safe boating practices, the Congress, by joint resolution approved June...

  5. Powering light boats

    Energy Technology Data Exchange (ETDEWEB)

    Affolter, J.-F.


    This short paper reports on a workshop held in Horw, Switzerland, on the application potential of low-temperature fuel cells. It looks at the increasing interest in small, electrically-driven boats in Europe and, in particular, at the option of powering them with fuel cells. A prototype implementation of a test boat is described. Powered by a 300-watt portable PEFC fuel cell constructed by the Paul Scherrer Institute (PSI) and the University of Applied Science in Solothurn, Switzerland, the boat is part of a project to study the technical feasibility of such drive systems for leisure craft and small boats. The paper addresses hydrogen security questions, energy-efficient drives and controllers, maintenance questions and discusses if a combination of fuel cells and photovoltaics would be viable. Also, the design of the boat's hull and the propellers that are used are looked at critically. The economic viability of fuel-cell powered boats is examined. The authors state that they are convinced that such fuel cell drives could be commercially interesting for larger passenger boats, too.

  6. Recreational Boating Statistics 2012 (United States)

    Department of Homeland Security — Every year, the USCG compiles statistics on reported recreational boating accidents. These statistics are derived from accident reports that are filed by the owners...

  7. Recreational Boating Statistics 2013 (United States)

    Department of Homeland Security — Every year, the USCG compiles statistics on reported recreational boating accidents. These statistics are derived from accident reports that are filed by the owners...

  8. In the Same Boat

    Institute of Scientific and Technical Information of China (English)



    Unlike the National Day or Spring Festival holidays that create massive logistical travel and traffic problems,the Dragon Boat Festival,falling on the fifth day of the fifth month of the Chinese lunar calendar or June 8 this year

  9. Dragon Boat Festival. (United States)

    Lew, Gordon

    This is one of a series of elementary readers written in Cantonese and English and designed to familiarize children with the traditional major Chinese festivals celebrated by the Chinese in America. This booklet describes the celebration of the Dragon Boat Festival, which marks the beginning of summer. A brief background to the festival is…

  10. Boats Floating Villas

    Institute of Scientific and Technical Information of China (English)


    <正>As China’s economy develops, motor hotels are being replaced gradually by boats, never becoming luxuries to many people as before.Imaging what you will feel when you are admired by the people standing to see you driving a motorboat along the lakeside and seaside, leaving white waves.

  11. Eleven cities Solar boat challenge

    NARCIS (Netherlands)

    Verdult, E.


    TU Delft students are participating for the fourth time in the world championship solar boat race in Friesland, which begins on 8 July. Expectations are high because the TU Delft students were world champions in 2006 and 2008. In 2010, the solar boat was equipped with hydrofoils for the first time.

  12. Row, Row, Row Your Boat...

    Institute of Scientific and Technical Information of China (English)



    @@ My afternoon plans to go dragon boat racing on Houhai Lake in downtown Beijing could have started better. Cycling across town, a pedal snapped off my bicycle as I was aggressively building up speed across a Second Ring Road junction.

  13. By boat to CERN

    CERN Multimedia


    Hello, my name is Jordan Juras. I will be here at CERN for a 2-month period spanning mid-June through mid-August working as Student Journalist for The Bulletin. This is a conventional position, but the manner with which I arrived at CERN is quite the opposite. Sitting at work one May 12th morning, I came across a Bulletin article (No. 20-21/2009) while perusing the LHC website. It was advertising ‘Fisica in Barca in Europe 2009’. They were looking for physicists living along the Mediterranean coast between Lisbon and Genoa to help out with the programme. Though I do not hold the title of physicist, nor do I live in Europe (in fact I live just outside of Harrow, Ontario, Canada), I felt compelled to write an email, expressing my love for physics and my interest in helping out with the programme. To my surprise, my help on board was welcome, and I managed to find some discount airfare and I set out for the South of Italy to meet the boat in Bari. Between Bari and Tries...

  14. Alternative Design of Boat Fenders

    DEFF Research Database (Denmark)

    Banke, Lars


    On offshore platforms the purpose of fenders is to protect the oil-risers against minor accidental collisions with supply vessels. Normally, the fender is designed by use of thin-walled tubes. However, the tube itself is not capable of resisting the impact load of the boat. Therefore, alternative...

  15. Peptide-LNA oligonucleotide conjugates

    DEFF Research Database (Denmark)

    Astakhova, I Kira; Hansen, Lykke Haastrup; Vester, Birte


    properties, peptides were introduced into oligonucleotides via a 2'-alkyne-2'-amino-LNA scaffold. Derivatives of methionine- and leucine-enkephalins were chosen as model peptides of mixed amino acid content, which were singly and doubly incorporated into LNA/DNA strands using highly efficient copper......Although peptide-oligonucleotide conjugates (POCs) are well-known for nucleic acids delivery and therapy, reports on internal attachment of peptides to oligonucleotides are limited in number. To develop a convenient route for preparation of internally labeled POCs with improved biomedical......(i)-catalyzed azide-alkyne cycloaddition (CuAAC) "click" chemistry. DNA/RNA target binding affinity and selectivity of the resulting POCs were improved in comparison to LNA/DNA mixmers and unmodified DNA controls. This clearly demonstrates that internal attachment of peptides to oligonucleotides can significantly...

  16. 78 FR 53156 - Sport Fishing and Boating Partnership Council; Teleconference (United States)


    ... Fish and Wildlife Service Sport Fishing and Boating Partnership Council; Teleconference AGENCY: Fish... Wildlife Service (Service), announce a public teleconference of the Sport Fishing and Boating Partnership.... App., we announce that Sport Fishing and Boating Partnership Council will hold a...

  17. Thermodynamics of Oligonucleotide Duplex Melting (United States)

    Schreiber-Gosche, Sherrie; Edwards, Robert A.


    Melting temperatures of oligonucleotides are useful for a number of molecular biology applications, such as the polymerase chain reaction (PCR). Although melting temperatures are often calculated with simplistic empirical equations, application of thermodynamics provides more accurate melting temperatures and an opportunity for students to apply…

  18. Cardboard Boat Building in Math Class (United States)

    Omundsen, John


    If you want to get the attention of a group of eighth grade math students, tell them they are going to build a life-size cardboard boat. To increase interest, follow up this statement by telling them that two to four of them will actually be rowing this boat across a small pond. Eighth grade math students at Oasis Charter Middle School in…

  19. Swiss fuel cell passenger and pleasure boats

    Energy Technology Data Exchange (ETDEWEB)

    Affolter, J.-F.


    This paper published by the University of Applied Science in Yverdon-les-Bains, Switzerland, looks at the development of electrically driven small boats that are powered by fuel cells. The various implementations of the test boats are described. Starting with a 100-watt PEM fuel cell built by the Paul Scherrer Institute (PSI) and the University of Applied Science in Solothurn, Switzerland, for educational purposes, a small pedal-boat was electrified. The paper describes the development of four further prototypes and introduces a new project for a 6-passenger leisure boat powered by a 2 kW PEFC fuel cell. Apart from the fuel cells, various other components such as propellers and control electronics are discussed as are the remaining problems still to be solved before the cells and boats can be marketed. Since they were carried out at a technical university, these projects are said to have provided an excellent way of teaching new technologies to students.

  20. 50 CFR 86.13 - What is boating infrastructure? (United States)


    ... (CONTINUED) FINANCIAL ASSISTANCE-WILDLIFE SPORT FISH RESTORATION PROGRAM BOATING INFRASTRUCTURE GRANT (BIG) PROGRAM General Information About the Grant Program § 86.13 What is boating infrastructure? Boating... 50 Wildlife and Fisheries 6 2010-10-01 2010-10-01 false What is boating infrastructure?...

  1. Small-boat Cetacean Surveys Database (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — The database contains multiple spreadsheets that hold data collected during each small-boat survey project conducted by the PIFSC CRP. This includes a summary of the...

  2. Marianas Boat Fishing Cost Earnings Study 2011 (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — These data represent a cost-earnings study of the boat based fishing in the Mariana Archipelago fielded in 2011. Data collected include fisher classification, vessel...

  3. Improvement and Validation of the BOAT Algorithm

    Directory of Open Access Journals (Sweden)

    Yingchun Liu


    Full Text Available The main objective of this paper is improving the BOAT classification algorithm and applying it in credit card big data analysis. Decision tree algorithm is a data analysis method for the classification which can be used to describe the extract important data class models or predict future data trends. The BOAT algorithm can reduce the data during reading and writing the operations, the improved algorithms in large data sets under the operating efficiency, and in line with the popular big data analysis. Through this paper, BOAT algorithm can further improve the performance of the algorithm and the distributed data sources under the performance. In this paper, large banking sectors of credit card data as the being tested data sets. The improved algorithm, the original BOAT algorithms, and the performance of other classical classification algorithms will be compared and analyzed.

  4. Oligonucleotide therapeutics: chemistry, delivery and clinical progress. (United States)

    Sharma, Vivek K; Watts, Jonathan K


    Oligonucleotide therapeutics have the potential to become a third pillar of drug development after small molecules and protein therapeutics. However, the three approved oligonucleotide drugs over the past 17 years have not proven to be highly successful in a commercial sense. These trailblazer drugs have nonetheless laid the foundations for entire classes of drug candidates to follow. This review will examine further advances in chemistry that are earlier in the pipeline of oligonucleotide drug candidates. Finally, we consider the possible effect of delivery systems that may provide extra footholds to improve the potency and specificity of oligonucleotide drugs. Our overview focuses on strategies to imbue antisense oligonucleotides with more drug-like properties and their applicability to other nucleic acid therapeutics.

  5. Adaptive resolution simulation of oligonucleotides (United States)

    Netz, Paulo A.; Potestio, Raffaello; Kremer, Kurt


    Nucleic acids are characterized by a complex hierarchical structure and a variety of interaction mechanisms with other molecules. These features suggest the need of multiscale simulation methods in order to grasp the relevant physical properties of deoxyribonucleic acid (DNA) and RNA using in silico experiments. Here we report an implementation of a dual-resolution modeling of a DNA oligonucleotide in physiological conditions; in the presented setup only the nucleotide molecule and the solvent and ions in its proximity are described at the atomistic level; in contrast, the water molecules and ions far from the DNA are represented as computationally less expensive coarse-grained particles. Through the analysis of several structural and dynamical parameters, we show that this setup reliably reproduces the physical properties of the DNA molecule as observed in reference atomistic simulations. These results represent a first step towards a realistic multiscale modeling of nucleic acids and provide a quantitatively solid ground for their simulation using dual-resolution methods.

  6. 78 FR 30723 - National Safe Boating Week, 2013 (United States)


    ... season. Safe boating starts onshore. Americans planning to spend a day on the water should prepare by... many lives every year, individuals should never operate a boat under the influence of drugs or...

  7. Tour Group Cruises the Three Gorges in Luxury Boat

    Institute of Scientific and Technical Information of China (English)


    BOARDING "Sheena," a luxury boat owned by the Sino-American Deluxe Boat Company, I began a five-day tour to Sanxia (Three Gorges of the Changjiang River) with ten other European and American tourists.

  8. Traditional boat-building and navigational techniques of southern Orissa

    Digital Repository Service at National Institute of Oceanography (India)

    Tripati, S.

    of the region. No written records on ancient boatbuilding and navigational aids of the region are available for the reconstruction of the technique of boat-building. Boats of this area have been classified into two categories, namely planked and log boats...

  9. 21 CFR 1240.95 - Sanitation of water boats. (United States)


    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Sanitation of water boats. 1240.95 Section 1240.95... DISEASES Source and Use of Potable Water § 1240.95 Sanitation of water boats. No vessel engaged in interstate traffic shall obtain water for drinking and culinary purposes from any water boat unless the...

  10. 75 FR 22423 - Sport Fishing and Boating Partnership Council (United States)


    ... Fish and Wildlife Service Sport Fishing and Boating Partnership Council AGENCY: Fish and Wildlife... public meeting of the Sport Fishing and Boating Partnership Council (Council). DATES: The meeting will be... announce that the Sport Fishing and Boating Partnership Council will hold a meeting. Background The...

  11. 78 FR 4161 - Sport Fishing and Boating Partnership Council (United States)


    ... Fish and Wildlife Service Sport Fishing and Boating Partnership Council AGENCY: Fish and Wildlife... (Service), announce a public teleconference of the Sport Fishing and Boating Partnership Council (Council.... App., we announce that Sport Fishing and Boating Partnership Council will hold a...

  12. 76 FR 7579 - Sport Fishing and Boating Partnership Council (United States)


    ... Fish and Wildlife Service Sport Fishing and Boating Partnership Council AGENCY: Fish and Wildlife... public meeting of the Sport Fishing and Boating Partnership Council (Council). DATES: The meeting will be... Federal Advisory Committee Act, 5 U.S.C. App., we announce that the Sport Fishing and Boating...

  13. 75 FR 47624 - Sport Fishing and Boating Partnership Council (United States)


    ... Fish and Wildlife Service Sport Fishing and Boating Partnership Council AGENCY: Fish and Wildlife... public meeting of the Sport Fishing and Boating Partnership Council (Council). DATES: The meeting will be.... App., we announce that the Sport Fishing and Boating Partnership Council will hold a meeting...

  14. 76 FR 1628 - Sport Fishing and Boating Partnership Council (United States)


    ... Fish and Wildlife Service Sport Fishing and Boating Partnership Council AGENCY: Fish and Wildlife...), announce a public teleconference of the Sport Fishing and Boating Partnership Council (Council). DATE(S... private sectors of the sport fishing, boating, and conservation communities and is organized to...

  15. 78 FR 33856 - Sport Fishing and Boating Partnership Council (United States)


    ...; FF09X60000-FVWF97920900000-XXX] Sport Fishing and Boating Partnership Council AGENCY: Fish and Wildlife... (Service), announce a public teleconference of the Sport Fishing and Boating Partnership Council (Council... Act, 5 U.S.C. App., we announce that Sport Fishing and Boating Partnership Council will hold...

  16. 77 FR 61626 - Sport Fishing and Boating Partnership Council (United States)


    ... Fish and Wildlife Service Sport Fishing and Boating Partnership Council AGENCY: Fish and Wildlife...), announce a public meeting of the Sport Fishing and Boating Partnership Council (Council). A Federal.... App., we announce that the Sport Fishing and Boating Partnership Council will hold a...

  17. 77 FR 26784 - Sport Fishing and Boating Partnership Council (United States)


    ... Fish and Wildlife Service Sport Fishing and Boating Partnership Council AGENCY: Fish and Wildlife... (Service), announce a public teleconference of the Sport Fishing and Boating Partnership Council (Council.... App., we announce that Sport Fishing and Boating Partnership Council will hold a...

  18. Recreational Boating Accidents and the Contribution of Boating Under the Influence (United States)

    Department of Homeland Security — This report includes statistical information for the fifty states, five U.S. territories, and the District of Columbia on recreational boating accident data and the...

  19. Delta Lloyd solar boat team; sailing 30km/h with a solar-powered boat, using hydrofoils

    NARCIS (Netherlands)

    Tra, M.


    Every two years since 2006, the Delta Lloyd Solar Boat Team builds a boat to participate in the World Cup for solar-powered boats, across the ‘Elfsteden-route’, the famous tour of the eleven cities in the Dutch province of Friesland. The upcoming year the team, consisting of a new group of students,

  20. Dragon Boat Festival (Dyun Ngh Jit). (United States)

    Wu, Julia; Quan, Ella Y.

    This bilingual-bicultural reader in Cantonese and English is intended for elementary school children in a bilingual education setting. Pen-and-ink drawings illustrate the story of the traditional dragon boat festival. Each page of the text is written in Chinese characters, Romanized form, and in English. (NCR)

  1. Kinetic effects on signal normalization in oligonucleotide microchips with labeled immobilized probes. (United States)

    Pan'kov, S V; Chechetkin, V R; Somova, O G; Antonova, O V; Moiseeva, O V; Prokopenko, D V; Yurasov, R A; Gryadunov, D A; Chudinov, A V


    Among various factors affecting operation of oligonucleotide microchips, the variations in concentration and in homogeneous distribution of immobilized probes over the cells are one of the most important. The labeling of immobilized probes ensures the complete current monitoring on the probe distribution and is reliable and convenient. Using hydrogel-based oligonucleotide microchips, the applicability of Cy3-labeled immobilized probes for quality control and signal normalization after hybridization with Cy5-labeled target DNA was investigated. This study showed that proper signal normalization should be different in thermodynamic conditions and in transient regime with hybridization far from saturation. This kinetic effect holds for both hydrogel-based and surface oligonucleotide microchips. Besides proving basic features, the technique was assessed on a sampling batch of 50 microchips developed for identifying mutations responsible for rifampicin and isoniazid resistance of Mycobacterium tuberculosis.

  2. An oligonucleotide hybridization approach to DNA sequencing. (United States)

    Khrapko, K R; Lysov YuP; Khorlyn, A A; Shick, V V; Florentiev, V L; Mirzabekov, A D


    We have proposed a DNA sequencing method based on hybridization of a DNA fragment to be sequenced with the complete set of fixed-length oligonucleotides (e.g., 4(8) = 65,536 possible 8-mers) immobilized individually as dots of a 2-D matrix [(1989) Dokl. Akad. Nauk SSSR 303, 1508-1511]. It was shown that the list of hybridizing octanucleotides is sufficient for the computer-assisted reconstruction of the structures for 80% of random-sequence fragments up to 200 bases long, based on the analysis of the octanucleotide overlapping. Here a refinement of the method and some experimental data are presented. We have performed hybridizations with oligonucleotides immobilized on a glass plate, and obtained their dissociation curves down to heptanucleotides. Other approaches, e.g., an additional hybridization of short oligonucleotides which continuously extend duplexes formed between the fragment and immobilized oligonucleotides, should considerably increase either the probability of unambiguous reconstruction, or the length of reconstructed sequences, or decrease the size of immobilized oligonucleotides.

  3. Archetypal Analysis of The Open Boat

    Institute of Scientific and Technical Information of China (English)



      The Open Boat is written by Stephen Crane according to his own experience of encountering a shipwreck. This short nov⁃el is not only a legend, but also a complex of the author’s thinking of life and human and a fable of men’s lives. It is full of symbols and metaphors. To interpret the meaning of this novel, this paper is engaged to have an archetypal analysis of this short story.

  4. Guam's Small-Boat-based Fisheries


    Myers, Robert F.


    Recent trends in Guam's small-boat fisheries and current knowledge of their biology, management, and economics are summarized. Annual estimates of participation, effort, and harvest are given for the pelagic and bottomfish fisheries for 1980-91 and for the spearfishing and atulai fisheries for 1985. The pelagic fishery is the largest, with annual landings ranging from 168 to 364 metric tons (t), followed by the bottomfish fishery (14-43 t), spearfishingf ishery (517 t), and bigeye scad fisher...

  5. Synthesis and hybridization property of a boat-shaped pyranosyl nucleic acid containing an exocyclic methylene group in the sugar moiety. (United States)

    Mori, Kazuto; Kodama, Tetsuya; Obika, Satoshi


    A boat-shaped pyranosyl nucleic acid (BsNA) having an exocyclic methylene group in the sugar moiety was synthesized to investigate the possibility that the axial H3' of original BsNA is the cause of its duplex destabilization. The synthesized BsNA analog was chemically stable against various nucleophiles. From the thermal stability of duplex oligonucleotides including the BsNA analog, it was found that the duplex-forming ability can be sensitive to the size of functional groups at the 3'-position.

  6. Chemosensitization by antisense oligonucleotides targeting MDM2. (United States)

    Bianco, Roberto; Ciardiello, Fortunato; Tortora, Giampaolo


    The MDM2 oncogene is overexpressed in many human cancers, including sarcomas, certain hematologic malignancies, and breast, colon and prostate cancers. The p53-MDM2 interaction pathway has been suggested as a novel target for cancer therapy. To that end, several strategies have been explored, including the use of small polypeptides targeted to the MDM2-p53 binding domain, anti-MDM2 antisense oligonucleotides, and natural agents. Different generations of anti-human-MDM2 oligonucleotides have been tested in in vitro and in vivo human cancer models, revealing specific inhibition of MDM2 expression and significant antitumor activity. Use of antisense oligos potentiated the effects of growth inhibition, p53 activation and p21 induction by several chemotherapeutic agents. Increased therapeutic effectiveness of chemotherapeutic drugs in human cancer cell lines carrying p53 mutations or deletions have shown the ability of MDM2 inhibitors to act as chemosensitizers in various types of tumors through both p53-dependent and p53-independent mechanisms. Inhibiting MDM2 appears to also have a role in radiation therapy for human cancer, regardless of p53 status, providing a rationale for the development of a new class of radiosensitizers. Moreover, MDM2 antisense oligonucleotides potentiate the effect of epidermal growth factor receptor (EGFR) inhibitors by affecting in vitro and in vivo proliferation, apoptosis and protein expression in hormone-refractory and hormone-dependent human prostate cancer cells. These data support the development, among other MDM2 inhibitors, of anti-MDM2 antisense oligonucleotides as a novel class of anticancer agents, and suggest a potentially relevant role for the oligonucleotides when integrated with conventional treatments and/or other signaling inhibitors in novel therapeutic strategies.

  7. Radio-controlled boat for measuring water velocities and bathymetry (United States)

    Vidmar, Andrej; Bezak, Nejc; Sečnik, Matej


    Radio-controlled boat named "Hi3" was designed and developed in order to facilitate water velocity and bathymetry measurements. The boat is equipped with the SonTek RiverSurveyor M9 instrument that is designed for measuring open channel hydraulics (discharge and bathymetry). Usually channel cross sections measurements are performed either from a bridge or from a vessel. However, these approaches have some limitations such as performing bathymetry measurements close to the hydropower plant turbine or downstream from a hydropower plant gate where bathymetry changes are often the most extreme. Therefore, the radio-controlled boat was designed, built and tested in order overcome these limitations. The boat is made from a surf board and two additional small balance support floats. Additional floats are used to improve stability in fast flowing and turbulent parts of rivers. The boat is powered by two electric motors, steering is achieved with changing the power applied to left and right motor. Furthermore, remotely controlled boat "Hi3" can be powered in two ways, either by a gasoline electric generator or by lithium batteries. Lithium batteries are lighter, quieter, but they operation time is shorter compared to an electrical generator. With the radio-controlled boat "Hi3" we can perform measurements in potentially dangerous areas such as under the lock gates at hydroelectric power plant or near the turbine outflow. Until today, the boat "Hi3" has driven more than 200 km in lakes and rivers, performing various water speed and bathymetry measurements. Moreover, in future development the boat "Hi3" will be upgraded in order to be able to perform measurements automatically. The future plans are to develop and implement the autopilot. With this approach the user will define the route that has to be driven by the boat and the boat will drive the pre-defined route automatically. This will be possible because of the very accurate differential GPS from the Sontek River

  8. A study of traditional boats of Goa

    Digital Repository Service at National Institute of Oceanography (India)

    Shaikh, Z.A.; Tripati, S.; Shinde, V.

    and species and sell in market. This shows the division of labour among the ‘kharvi’ community. ‘Ramponkars’ also maintain a book in which apart from profit and loss they also record who were participated in the catch. Further, 10% of the profit is kept...., and Kentley, E., 1985. Sewn-Plank Boats. Oxford: British Archaeological Reports. McGrail, S. 1996. Towards a Classification of Water Transport, In Marine Archaeology the Global Perspectives (G. Kuppuram and K. Kumudamani, Eds.), pp. 187-204. New Delhi...

  9. 78 FR 18617 - Recreational Boating Safety Projects, Programs and Activities Funded Under Provisions of the... (United States)


    ... National Recreational Boating Safety Program Strategic Plan. ($23,367). Boating Accident News Clipping... Program: Funding was allocated for this program, which provides full marketing, media, public...

  10. Louisiana Marinas and Boat Launches, Geographic NAD83, LOSCO (2004) [marinas_LOSCO_2004 (United States)

    Louisiana Geographic Information Center — The dataset defines the location and supplemental information for marinas and boat launches in southern Louisiana. The boat launch database includes public and...

  11. Disentangling invasion processes in a dynamic shipping-boating network. (United States)

    Lacoursière-Roussel, Anaïs; Bock, Dan G; Cristescu, Melania E; Guichard, Frédéric; Girard, Philippe; Legendre, Pierre; McKindsey, Christopher W


    The relative importance of multiple vectors to the initial establishment, spread and population dynamics of invasive species remains poorly understood. This study used molecular methods to clarify the roles of commercial shipping and recreational boating in the invasion by the cosmopolitan tunicate, Botryllus schlosseri. We evaluated (i) single vs. multiple introduction scenarios, (ii) the relative importance of shipping and boating to primary introductions, (iii) the interaction between these vectors for spread (i.e. the presence of a shipping-boating network) and (iv) the role of boating in determining population similarity. Tunicates were sampled from 26 populations along the Nova Scotia, Canada, coast that were exposed to either shipping (i.e. ports) or boating (i.e. marinas) activities. A total of 874 individuals (c. 30 per population) from five ports and 21 marinas was collected and analysed using both mitochondrial cytochrome c oxidase subunit I gene (COI) and 10 nuclear microsatellite markers. The geographical location of multiple hotspot populations indicates that multiple invasions have occurred in Nova Scotia. A loss of genetic diversity from port to marina populations suggests a stronger influence of ships than recreational boats on primary coastal introductions. Population genetic similarity analysis reveals a dependence of marina populations on those that had been previously established in ports. Empirical data on marina connectivity because of boating better explains patterns in population similarities than does natural spread. We conclude that frequent primary introductions arise by ships and that secondary spread occurs gradually thereafter around individual ports, facilitated by recreational boating.

  12. 77 FR 33229 - Sport Fishing and Boating Partnership Council Charter (United States)


    ... Office of the Secretary Sport Fishing and Boating Partnership Council Charter AGENCY: Office of the... ] consultation with the General Services Administration, the Secretary of the Interior has renewed the Sport... or programs to increase public awareness and support for the Sport Fish Restoration and Boating...

  13. 76 FR 75898 - Sport Fishing and Boating Partnership Council (United States)


    ... Fish and Wildlife Service Sport Fishing and Boating Partnership Council AGENCY: Fish and Wildlife... (Service), announce a public teleconference of the Sport Fishing and Boating Partnership Council (Council.... The Council represents the interests of the public and private sectors of the sport fishing,...

  14. 77 FR 29359 - Sport Fishing and Boating Partnership Council (United States)


    ... Fish and Wildlife Service Sport Fishing and Boating Partnership Council AGENCY: Fish and Wildlife...), announce a public meeting of the Sport Fishing and Boating Partnership Council (Council). A Federal... the requirements of the Federal Advisory Committee Act, 5 U.S.C. App., we announce that the...

  15. 76 FR 47606 - Sport Fishing and Boating Partnership Council (United States)


    ... Fish and Wildlife Service Sport Fishing and Boating Partnership Council AGENCY: Fish and Wildlife... public meeting of the Sport Fishing and Boating Partnership Council (Council). DATES: The meeting will be... of the Federal Advisory Committee Act, 5 U.S.C. App., we announce that the Sport Fishing and...

  16. 78 FR 23587 - Sport Fishing and Boating Partnership Council (United States)


    ... Fish and Wildlife Service Sport Fishing and Boating Partnership Council AGENCY: Fish and Wildlife...), announce a public meeting of the Sport Fishing and Boating Partnership Council (Council). A Federal... requirements of the Federal Advisory Committee Act, 5 U.S.C. App., we announce that the Sport Fishing...

  17. 76 FR 81959 - Sport Fishing and Boating Partnership Council (United States)


    ... Fish and Wildlife Service Sport Fishing and Boating Partnership Council AGENCY: Fish and Wildlife... public meeting of the Sport Fishing and Boating Partnership Council (Council). DATES: The meeting will be... with the requirements of the Federal Advisory Committee Act, 5 U.S.C. App., we announce that the...

  18. Interaction between Brash Ice and Boat Propulsion Systems (United States)


    seal it from water intrusion to the boat. The propulsion propeller was mounted mid-way between the end of the cut in the bottom of the boat and grown from saltwater using a methodology developed at CRREL by Hirayama (1983) for urea-doped water. The ice is grown in the test basin on the

  19. Automatic Boat Identification System for VIIRS Low Light Imaging Data

    Directory of Open Access Journals (Sweden)

    Christopher D. Elvidge


    Full Text Available The ability for satellite sensors to detect lit fishing boats has been known since the 1970s. However, the use of the observations has been limited by the lack of an automatic algorithm for reporting the location and brightness of offshore lighting features arising from boats. An examination of lit fishing boat features in Visible Infrared Imaging Radiometer Suite (VIIRS day/night band (DNB data indicates that the features are essentially spikes. We have developed a set of algorithms for automatic detection of spikes and characterization of the sharpness of spike features. A spike detection algorithm generates a list of candidate boat detections. A second algorithm measures the height of the spikes for the discard of ionospheric energetic particle detections and to rate boat detections as either strong or weak. A sharpness index is used to label boat detections that appear blurry due to the scattering of light by clouds. The candidate spikes are then filtered to remove features on land and gas flares. A validation study conducted using analyst selected boat detections found the automatic algorithm detected 99.3% of the reference pixel set. VIIRS boat detection data can provide fishery agencies with up-to-date information of fishing boat activity and changes in this activity in response to new regulations and enforcement regimes. The data can provide indications of illegal fishing activity in restricted areas and incursions across Exclusive Economic Zone (EEZ boundaries. VIIRS boat detections occur widely offshore from East and Southeast Asia, South America and several other regions.

  20. Antisense oligonucleotides in therapy for neurodegenerative disorders. (United States)

    Evers, Melvin M; Toonen, Lodewijk J A; van Roon-Mom, Willeke M C


    Antisense oligonucleotides are synthetic single stranded strings of nucleic acids that bind to RNA and thereby alter or reduce expression of the target RNA. They can not only reduce expression of mutant proteins by breakdown of the targeted transcript, but also restore protein expression or modify proteins through interference with pre-mRNA splicing. There has been a recent revival of interest in the use of antisense oligonucleotides to treat several neurodegenerative disorders using different approaches to prevent disease onset or halt disease progression and the first clinical trials for spinal muscular atrophy and amyotrophic lateral sclerosis showing promising results. For these trials, intrathecal delivery is being used but direct infusion into the brain ventricles and several methods of passing the blood brain barrier after peripheral administration are also under investigation.

  1. Electrochemical study of hepta–oligonucleotides

    Directory of Open Access Journals (Sweden)

    Zdenka Balcarova


    Full Text Available The study deals with the description and characterization of twohepta–oligonucleotides (DNA and RNA forming special structures.We studied their electrochemical behaviour by means of cyclicvoltammetry (CV and elimination voltammetry with linear scan(EVLS in combination with adsorptive stripping (AdS technique.Differences in electrochemical behaviour of hepta–deoxyribonucleotide and its RNA analog were discussed with regardto their different structures in solutions and their melting points.

  2. Synthesis and hybridization properties of inverse oligonucleotides.


    Marangoni, M.; Van Aerschot, Arthur; Augustijns, Patrick; Rozenski, Jef; Herdewijn , Piet


    The synthesis of adenine and thymine cyclopentylethyl nucleosides is presented. This novel constrained monomeric building block is very difficult to incorporate into oligonucleotides. It was introduced in 13mer oligodeoxynucleotide sequences at a single position using H-phosphonate chemistry. Phosphoramidite chemistry completely failed in this particular case. The H-phosphonate building blocks were obtained starting from the corresponding phosphoramidites. Stability of duplexes with RNA and D...

  3. Flight of the loon : solar-powered boating

    Energy Technology Data Exchange (ETDEWEB)



    Details of the first solar-powered, zero-emission boat to venture across the Trent-Severn waterway were presented. Called the 'Loon', the boat is now being manufactured for environmentally-conscious boaters around the world. The seating arrangement, pontoon design, and propulsion system were all designed to minimize impacts on the ecosystem. A prototype boat was built and tested in the summer of 2005 to determine the viability of the solar-powered boat, and a family trip down the waterway was completed in August. Results of the trip encouraged the inventor to raise capital and a suitable production facility. After several months of development, the first production boat was completed with many improvements to the original prototype. An entrepreneur from Belize was the first person to place an order after he discovered that most commercially-available solar-powered boats were higher-priced and less capable than the Loon. High gas prices and the detrimental effects of fuel exhaust in lakes have caused many people to become sensitive to the issues surrounding fuel use and pleasure boats. It was concluded that a storefront has recently been opened north of Orillia in order to sell the solar-powered craft. 3 figs.

  4. The Boat Girl in the Photo

    Institute of Scientific and Technical Information of China (English)


    The Little Boat Girl THIS picture was taken at dusk, onestormy day, half a century ago. On April 21, 1949, one million soldiersof the Chinese People’s Liberation Army, alongmore than 500-kilometers of riverbank, quicklybroke through the "thousand-Li line of defence,"which had been painstakingly built up by theKuomintang, and forced the Changjiang River. At dusk on April 22, boatmen from Yangzhong,an island in the Changjiang River, hurried to thebank to row the follow-up units across. Despitearduous weather, the boatmen’s skill in fightingagainst wind and waves was considerable and, inthe soft lights of sunset, the rows of old woodenboats appeared to be flying on the ChangjiangRiver. A most impressive scene. Among the officers waiting on the riverbank tobe ferried across was one tall soldier, deeplyabsorbed in the sight of a young girl, not far away,

  5. Template-Directed Ligation of Peptides to Oligonucleotides (United States)

    Bruick, Richard K.; Dawson, Philip E.; Kent, Stephen BH; Usman, Nassim; Joyce, Gerald F.


    Synthetic oligonucleotides and peptides have enjoyed a wide range of applications in both biology and chemistry. As a consequence, oligonucleotide-peptide conjugates have received considerable attention, most notably in the development of antisense constructs with improved pharmacological properties. In addition, oligonucleotide-peptide conjugates have been used as molecular tags, in the assembly of supramolecular arrays and in the construction of encoded combinatorial libraries. To make these chimeric molecules more accessible for a broad range of investigations, we sought to develop a facile method for joining fully deprotected oligonucleotides and peptides through a stable amide bond linkage. Furthermore, we wished to make this ligation reaction addressable, enabling one to direct the ligation of specific oligonucleotide and peptide components.To confer specificity and accelerate the rate of the reaction, the ligation process was designed to be dependent on the presence of a complementary oligonucleotide template.

  6. Patrol Vessels «Condestable Zaragoza» Class, Patrol Boats V-class and Antisubmarine Boats LAS-class

    Directory of Open Access Journals (Sweden)

    Valentin Ruesga Herreros


    Full Text Available It’s consider information about the number of ships of the Spanish fleet, that not reflected in the available literature. It’s proved that the Spanish patrol vessels of «Condestable Zaragoza»-class and French auxiliary minesweepers «Fanfaron»-classare one the same ships. The conclusion is made based on comparisons of appearance and performance of ships. There was renovated list of patrol boats V-class, shows that their history can be divided into three main stages: the first, when the boats belonged to the Secretariat militarized protection, information about boats are extremely sketchy and incomplete, the second – their inclusion in the fleet, when the information about the service enough full, but almost previous history is not reflected, and only the third stage, when the change came a new generation of boats old boats, often with the same numbers, the story is full enough. An attempt to reconstruction of the biography of LAS-class boats, it is assumed that this former Soviet boat, put the Republicans during the Civil War.

  7. Hawaii Small Boat Cost-Earnings Data: 2007-2008 (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — These data represent a cost-earnings study of the Hawaii small boat fishery in 2007-2008. Data collected include fisher classification, vessel characteristics,...

  8. Hawaii Small Boat Cost-Earnings Data: 2014 (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — These data represent a cost-earnings study of the Hawaii small boat fishery in 2014. Data collected include fisher classification, vessel characteristics, levels of...

  9. Hawaii Small Boat Cost-Earnings Data: 1995-1996 (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — These data represent a cost-earnings study of the Hawaii small boat fishery in 1995-1996. Data collected include fisher classification, vessel characteristics,...

  10. Template switching between PNA and RNA oligonucleotides (United States)

    Bohler, C.; Nielsen, P. E.; Orgel, L. E.; Miller, S. L. (Principal Investigator)


    The origin of the RNA world is not easily understood, as effective prebiotic syntheses of the components of RNA, the beta-ribofuranoside-5'-phosphates, are hard to envisage. Recognition of this difficulty has led to the proposal that other genetic systems, the components of which are more easily formed, may have preceded RNA. This raises the question of how transitions between one genetic system and another could occur. Peptide nucleic acid (PNA) resembles RNA in its ability to form double-helical complexes stabilized by Watson-Crick hydrogen bonding between adenine and thymine and between cytosine and guanine, but has a backbone that is held together by amide rather than by phosphodiester bonds. Oligonucleotides bases on RNA are known to act as templates that catalyse the non-enzymatic synthesis of their complements from activated mononucleotides, we now show that RNA oligonucleotides facilitate the synthesis of complementary PNA strands and vice versa. This suggests that a transition between different genetic systems can occur without loss of information.

  11. Enhanced fluorescence of silver nanoclusters stabilized with branched oligonucleotides. (United States)

    Latorre, Alfonso; Lorca, Romina; Zamora, Félix; Somoza, Álvaro


    DNA stabilized silver nanoclusters (AgNCs) are promising optical materials, whose fluorescence properties can be tuned by the selection of the DNA sequence employed. In this work we have used modified oligonucleotides in the preparation of AgNCs. The fluorescent intensity obtained was 60 times higher than that achieved with standard oligonucleotides.

  12. Noncoding oligonucleotides: the belle of the ball in gene therapy. (United States)

    Shum, Ka-To; Rossi, John J


    Gene therapy carries the promise of cures for many diseases based on manipulating the expression of a person's genes toward the therapeutic goal. The relevance of noncoding oligonucleotides to human disease is attracting widespread attention. Noncoding oligonucleotides are not only involved in gene regulation, but can also be modified into therapeutic tools. There are many strategies that leverage noncoding oligonucleotides for gene therapy, including small interfering RNAs, antisense oligonucleotides, aptamers, ribozymes, decoys, and bacteriophage phi 29 RNAs. In this chapter, we will provide a broad, comprehensive overview of gene therapies that use noncoding oligonucleotides for disease treatment. The mechanism and development of each therapeutic will be described, with a particular focus on its clinical development. Finally, we will discuss the challenges associated with developing nucleic acid therapeutics and the prospects for future success.

  13. Predicting oligonucleotide-directed mutagenesis failures in protein engineering. (United States)

    Wassman, Christopher D; Tam, Phillip Y; Lathrop, Richard H; Weiss, Gregory A


    Protein engineering uses oligonucleotide-directed mutagenesis to modify DNA sequences through a two-step process of hybridization and enzymatic synthesis. Inefficient reactions confound attempts to introduce mutations, especially for the construction of vast combinatorial protein libraries. This paper applied computational approaches to the problem of inefficient mutagenesis. Several results implicated oligonucleotide annealing to non-target sites, termed 'cross-hybridization', as a significant contributor to mutagenesis reaction failures. Test oligonucleotides demonstrated control over reaction outcomes. A novel cross-hybridization score, quickly computable for any plasmid and oligonucleotide mixture, directly correlated with yields of deleterious mutagenesis side products. Cross-hybridization was confirmed conclusively by partial incorporation of an oligonucleotide at a predicted cross-hybridization site, and by modification of putative template secondary structure to control cross-hybridization. Even in low concentrations, cross-hybridizing species in mixtures poisoned reactions. These results provide a basis for improved mutagenesis efficiencies and increased diversities of cognate protein libraries.

  14. Conjugation of fluorescent proteins with DNA oligonucleotides. (United States)

    Lapiene, Vidmantas; Kukolka, Florian; Kiko, Kathrin; Arndt, Andreas; Niemeyer, Christof M


    This work describes the synthesis of covalent ssDNA conjugates of six fluorescent proteins, ECFP, EGFP, E(2)GFP, mDsRed, Dronpa, and mCherry, which were cloned with an accessible C-terminal cystein residue to enable site-selective coupling using a heterobispecific cross-linker. The resulting conjugates revealed similar fluorescence emission intensity to the unconjugated proteins, and the functionality of the tethered oligonucleotide was proven by specific Watson-Crick base pairing to cDNA-modified gold nanoparticles. Fluorescence spectroscopy analysis indicated that the fluorescence of the FP is quenched by the gold particle, and the extent of quenching varied with the intrinsic spectroscopic properties of FP as well as with the configuration of surface attachment. Since this study demonstrates that biological fluorophores can be selectively incorporated into and optically coupled with nanoparticle-based devices, applications in DNA-based nanofabrication can be foreseen.

  15. Asthma Basics (United States)

    ... Old Feeding Your 1- to 2-Year-Old Asthma Basics KidsHealth > For Parents > Asthma Basics A A ... Asthma Categories en español Asma: aspectos fundamentales About Asthma Asthma is a common lung condition in kids ...

  16. Preparation and application of triple helix forming oligonucleotides and single strand oligonucleotide donors for gene correction. (United States)

    Alam, Rowshon; Thazhathveetil, Arun Kalliat; Li, Hong; Seidman, Michael M


    Strategies for site-specific modulation of genomic sequences in mammalian cells require two components. One must be capable of recognizing and activating a specific target sequence in vivo, driving that site into an exploitable repair pathway. Information is transferred to the site via participation in the pathway by the second component, a donor nucleic acid, resulting in a permanent change in the target sequence. We have developed biologically active triple helix forming oligonucleotides (TFOs) as site-specific gene targeting reagents. These TFOs, linked to DNA reactive compounds (such as a cross-linking agent), activate pathways that can engage informational donors. We have used the combination of a psoralen-TFO and single strand oligonucleotide donors to generate novel cell lines with directed sequence changes at the target site. Here we describe the synthesis and purification of bioactive psoralen-linked TFOs, their co-introduction into mammalian cells with donor nucleic acids, and the identification of cells with sequence conversion of the target site. We have emphasized details in the synthesis and purification of the oligonucleotides that are essential for preparation of reagents with optimal activity.

  17. On Naturalism in Stephen Crane’s The Open Boat

    Institute of Scientific and Technical Information of China (English)

    WANG Xiu-hong; CHEN Bei-bei


    Stephen Crane is one of the greatest figures in the history of American literature and his novella The Open Boat is wide-ly known by people all over the world as a representative work of Naturalism. This paper aims at exploring Naturalism in Stephen Crane’s The Open Boat, which will guide the readers to understand American Naturalism accurately and thoroughly. It firstly in-troduces the author and his novel The Open Boat. Secondly, it progressively dissects Naturalism. It not only analyses the general characteristics of Naturalism with details in The Open Boat, but also interprets the distinguishing features of American Naturalism so that readers can better understand the features of American Naturalism. Through textual perusal, it further studies the unique characteristics of Stephen Crane’s Naturalism and lays particular emphasis on Crane’s special concern on the awakening of hu-manity. It is hoped that this thesis can help readers get an in-depth comprehension of Naturalism and gain some inspiration from The Open Boat.

  18. Nucleic acid sequence detection using multiplexed oligonucleotide PCR (United States)

    Nolan, John P.; White, P. Scott


    Methods for rapidly detecting single or multiple sequence alleles in a sample nucleic acid are described. Provided are all of the oligonucleotide pairs capable of annealing specifically to a target allele and discriminating among possible sequences thereof, and ligating to each other to form an oligonucleotide complex when a particular sequence feature is present (or, alternatively, absent) in the sample nucleic acid. The design of each oligonucleotide pair permits the subsequent high-level PCR amplification of a specific amplicon when the oligonucleotide complex is formed, but not when the oligonucleotide complex is not formed. The presence or absence of the specific amplicon is used to detect the allele. Detection of the specific amplicon may be achieved using a variety of methods well known in the art, including without limitation, oligonucleotide capture onto DNA chips or microarrays, oligonucleotide capture onto beads or microspheres, electrophoresis, and mass spectrometry. Various labels and address-capture tags may be employed in the amplicon detection step of multiplexed assays, as further described herein.

  19. Design and analysis of mismatch probes for long oligonucleotide microarrays

    Energy Technology Data Exchange (ETDEWEB)

    Deng, Ye; He, Zhili; Van Nostrand, Joy D.; Zhou, Jizhong


    Nonspecific hybridization is currently a major concern with microarray technology. One of most effective approaches to estimating nonspecific hybridizations in oligonucleotide microarrays is the utilization of mismatch probes; however, this approach has not been used for longer oligonucleotide probes. Here, an oligonucleotide microarray was constructed to evaluate and optimize parameters for 50-mer mismatch probe design. A perfect match (PM) and 28 mismatch (MM) probes were designed for each of ten target genes selected from three microorganisms. The microarrays were hybridized with synthesized complementary oligonucleotide targets at different temperatures (e.g., 42, 45 and 50 C). In general, the probes with evenly distributed mismatches were more distinguishable than those with randomly distributed mismatches. MM probes with 3, 4 and 5 mismatched nucleotides were differentiated for 50-mer oligonucleotide probes hybridized at 50, 45 and 42 C, respectively. Based on the experimental data generated from this study, a modified positional dependent nearest neighbor (MPDNN) model was constructed to adjust the thermodynamic parameters of matched and mismatched dimer nucleotides in the microarray environment. The MM probes with four flexible positional mismatches were designed using the newly established MPDNN model and the experimental results demonstrated that the redesigned MM probes could yield more consistent hybridizations. Conclusions: This study provides guidance on the design of MM probes for long oligonucleotides (e.g., 50 mers). The novel MPDNN model has improved the consistency for long MM probes, and this modeling method can potentially be used for the prediction of oligonucleotide microarray hybridizations.

  20. Antisense oligonucleotide induction of progerin in human myogenic cells.

    Directory of Open Access Journals (Sweden)

    Yue-Bei Luo

    Full Text Available We sought to use splice-switching antisense oligonucleotides to produce a model of accelerated ageing by enhancing expression of progerin, translated from a mis-spliced lamin A gene (LMNA transcript in human myogenic cells. The progerin transcript (LMNA Δ150 lacks the last 150 bases of exon 11, and is translated into a truncated protein associated with the severe premature ageing disease, Hutchinson-Gilford progeria syndrome (HGPS. HGPS arises from de novo mutations that activate a cryptic splice site in exon 11 of LMNA and result in progerin accumulation in tissues of mesodermal origin. Progerin has also been proposed to play a role in the 'natural' ageing process in tissues. We sought to test this hypothesis by producing a model of accelerated muscle ageing in human myogenic cells. A panel of splice-switching antisense oligonucleotides were designed to anneal across exon 11 of the LMNA pre-mRNA, and these compounds were transfected into primary human myogenic cells. RT-PCR showed that the majority of oligonucleotides were able to modify LMNA transcript processing. Oligonucleotides that annealed within the 150 base region of exon 11 that is missing in the progerin transcript, as well as those that targeted the normal exon 11 donor site induced the LMNA Δ150 transcript, but most oligonucleotides also generated variable levels of LMNA transcript missing the entire exon 11. Upon evaluation of different oligomer chemistries, the morpholino phosphorodiamidate oligonucleotides were found to be more efficient than the equivalent sequences prepared as oligonucleotides with 2'-O-methyl modified bases on a phosphorothioate backbone. The morpholino oligonucleotides induced nuclear localised progerin, demonstrated by immunostaining, and morphological nuclear changes typical of HGPS cells. We show that it is possible to induce progerin expression in myogenic cells using splice-switching oligonucleotides to redirect splicing of LMNA. This may offer a model


    Institute of Scientific and Technical Information of China (English)

    MA Cheng; QIAN Zheng-fang; DU Du; ZHANG Xu


    Wake energy recovery of a ship's propeller is an important research subject in ship propulsion field. Based on the study on the mechanism analysis and computation method of the hydrodynamic performance of the propeller-rudder-rudder bubble combination, computer simulation system was built and then used to design and simulate four kinds of rudder bubbles, then the model comparison tests with the original rudder was carried out and the sensible factors influencing the energy-saving effects of rudder bubble was also analyzed. On this basis, a further optimized design of the rudder bubble for a fishing boat was put forward, and comparison test on a real boat was then carried out. It is shown that the optimized rudder bubble design can effectively recover wake energy of ships and the rudder bubble installation features a simple operation and high cost-effectiveness, and the energy-saving effect of real boat test achieves 5.1%.

  2. Self-propulsion of a metallic superoleophobic micro-boat. (United States)

    Musin, Albina; Grynyov, Roman; Frenkel, Mark; Bormashenko, Edward


    The self-propulsion of a heavy, superoleophobic, metallic micro-boat carrying a droplet of various aqueous alcohol solutions as a fuel tank is reported. The micro-boat is driven by the solutocapillary Marangoni flow. The jump in the surface tension owing to the condensation of alcohols on the water surface was established experimentally. Maximal velocities of the self-propulsion were registered as high as 0.05m/s. The maximal velocity of the center mass of the boat correlates with the maximal change in the surface tension, due to the condensation of alcohols. The mechanism of the self-locomotion is discussed. The phenomenological dynamic model describing the self-propulsion is reported.

  3. Optical Characterization of Oligonucleotide DNA Influenced by Magnetic Fields

    Directory of Open Access Journals (Sweden)

    Seyedeh Maryam Banihashemian


    Full Text Available UV-VIS spectroscopic analysis of oligonucleotide DNA exposed to different magnetic fields was performed in order to investigate the relationship between DNA extinction coefficients and optical parameters according to magnetic-field strength. The results with the oligonucleotides adenine-thymine 100 mer (AT-100 DNA and cytosine-guanine 100 mer (CG-100 DNA indicate that the magnetic field influences DNA molar extinction coefficients and refractive indexes. The imaginary parts of the refractive index and molar extinction coefficients of the AT-100 and CG-100 DNA decreased after exposure to a magnetic field of 750 mT due to cleavage of the DNA oligonucleotides into smaller segments.

  4. Allyl group as a protecting group for internucleotide phosphate and thiophosphate linkages in oligonucleotide synthesis: facile oxidation and deprotection conditions. (United States)

    Manoharan, M; Lu, Y; Casper, M D; Just, G


    [reaction: see text] The allyl group, which serves as a protecting group for an internucleotide bond for both phosphates and phosphorothioates, can be easily removed by good nucleophiles under weakly basic or neutral conditions. For a practical synthesis on solid support, camphorsulfonyloxaziridine was used as the oxidizing agent for synthesizing DNA, while the Beaucage reagent was used for preparing phosphorothioate oligomers. Both types of oligonucleotides were easily deprotected by concentrated ammonium hydroxide containing 2% mercaptoethanol.

  5. Summary on China boat import and export in 2005

    Institute of Scientific and Technical Information of China (English)

    Zhu Huaixin


    @@ According to the latest statistics by the Customs, China exported inflatable recreation/ sports boats in 4 types (as the Customs codes 89039900, 89031000, 89039200& 89039100) increased in large numbers up to 137634 and totaled 63.4096 million US dollars, which increased 40.8% than 2004 and took 1.344% of the total ship export sum of 2005.Besides, it also imported 1162 various boats mounting to 18.108 million US dollars,this increased 80.5% than 2004 and took 3.69%of the total ship import value.

  6. Basic electrotechnology

    CERN Document Server

    Ashen, R A


    BASIC Electrotechnology discusses the applications of Beginner's All-purpose Symbolic Instruction Code (BASIC) in engineering, particularly in solving electrotechnology-related problems. The book is comprised of six chapters that cover several topics relevant to BASIC and electrotechnology. Chapter 1 provides an introduction to BASIC, and Chapter 2 talks about the use of complex numbers in a.c. circuit analysis. Chapter 3 covers linear circuit analysis with d.c. and sinusoidal a.c. supplies. The book also discusses the elementary magnetic circuit theory. The theory and performance of two windi

  7. 19 CFR 12.85 - Coast Guard boat and associated equipment safety standards. (United States)


    ... 19 Customs Duties 1 2010-04-01 2010-04-01 false Coast Guard boat and associated equipment safety... Associated Equipment § 12.85 Coast Guard boat and associated equipment safety standards. (a) Applicability of standards or regulations prescribed by the Commandant, U.S. Coast Guard. Boats and associated equipment...

  8. 7 CFR 58.317 - Bulk butter trucks, boats, texturizers, and packers. (United States)


    ... 7 Agriculture 3 2010-01-01 2010-01-01 false Bulk butter trucks, boats, texturizers, and packers... and Grading Service 1 Equipment and Utensils § 58.317 Bulk butter trucks, boats, texturizers, and packers. Bulk butter trucks, boats, texturizers, and packers shall be constructed of aluminum,...

  9. 75 FR 17754 - Certificate of Alternative Compliance for the Lift Boat GARY CHIASSON ELEVATOR (United States)


    ... SECURITY Coast Guard Certificate of Alternative Compliance for the Lift Boat GARY CHIASSON ELEVATOR AGENCY... Compliance was issued for the lift boat GARY CHIASSON ELEVATOR as required by 33 U.S.C. 1605(c) and 33 CFR 81... Regulations, Parts 81 and 89, has been issued for the lift boat GARY CHIASSON ELEVATOR. The Certificate...

  10. 76 FR 27284 - Special Local Regulation; Partnership in Education, Dragon Boat Race; Maumee River, Toledo, OH (United States)


    ..., Dragon Boat Race; Maumee River, Toledo, OH AGENCY: Coast Guard, DHS. ACTION: Notice of proposed... during the Dragon Boat Races which take place during the third or fourth weekend in July each year. This... from other vessel traffic. The Captain of the Port Detroit has determined dragon boat races in...

  11. 78 FR 24065 - Special Local Regulation; Tuscaloosa Dragon Boat Races; Black Warrior River; Tuscaloosa, AL (United States)


    ... SECURITY Coast Guard 33 CFR Part 100 RIN 1625-AA08 Special Local Regulation; Tuscaloosa Dragon Boat Races... the Junior League of Tuscaloosa Dragon Boat Races. Entry into, transiting or anchoring in this area is... the Junior League of Tuscaloosa Dragon Boat Races. This event is advertised as scheduled...

  12. 77 FR 55436 - Special Local Regulation; Partnership in Education, Dragon Boat Race; Maumee River, Toledo, OH (United States)


    ..., Dragon Boat Race; Maumee River, Toledo, OH AGENCY: Coast Guard, DHS. ACTION: Notice of proposed... River during the annual Dragon Boat Races. This special local regulated area is necessary to protect... Regulation; Partnership in Education Dragon Boat Race, Maumee River Toledo, OH in the Federal...

  13. 77 FR 23125 - Special Local Regulation; Tuscaloosa Dragon Boat Race; Black Warrior River; Tuscaloosa, AL (United States)


    ... SECURITY Coast Guard 33 CFR Part 100 RIN 1625-AA08 Special Local Regulation; Tuscaloosa Dragon Boat Race.... League of Tuscaloosa Dragon Boat Races. Entry into, transiting or anchoring in this area is prohibited to... associated with the Jr. League of Tuscaloosa Dragon Boat Races. This event is advertised as scheduled...

  14. 78 FR 57063 - Special Local Regulations; Jacksonville Dragon Boat Festival; St. Johns River; Jacksonville, FL (United States)


    ... SECURITY Coast Guard 33 CFR Part 100 RIN 1625-AA08 Special Local Regulations; Jacksonville Dragon Boat... Jacksonville, Florida during the Jacksonville Dragon Boat Festival, a series of paddle boat races. The event is... is to ensure safety of life on navigable waters of the United States during the Jacksonville...

  15. 19 CFR 148.32 - Vehicles, aircraft, boats, teams and saddle horses taken abroad. (United States)


    ... 19 Customs Duties 2 2010-04-01 2010-04-01 false Vehicles, aircraft, boats, teams and saddle horses... for Returning Residents § 148.32 Vehicles, aircraft, boats, teams and saddle horses taken abroad. (a) Admission free of duty. Automobiles and other vehicles, aircraft, boats, teams and saddle horses,...

  16. Oligonucleotide and Long Polymeric DNA Encoding

    Energy Technology Data Exchange (ETDEWEB)

    Miller, E; Mariella Jr., R P; Christian, A T; Gardner, S N; Williams, J M


    This report summarizes the work done at Lawrence Livermore National Laboratory for the Oligonucleotide and Long Polymeric DNA Encoding project, part of the Microelectronic Bioprocesses Program at DARPA. The goal of the project was to develop a process by which long (circa 10,000 base-pair) synthetic DNA molecules could be synthesized in a timely and economic manner. During construction of the long molecule, errors in DNA sequence occur during hybridization and/or the subsequent enzymatic process. The work done on this project has resulted in a novel synthesis scheme that we call the parallel pyramid synthesis protocol, the development of a suit of computational tools to minimize and quantify errors in the synthesized DNA sequence, and experimental proof of this technique. The modeling consists of three interrelated modules: the bioinformatics code which determines the specifics of parallel pyramid synthesis for a given chain of long DNA, the thermodynamics code which tracks the products of DNA hybridization and polymerase extension during the later steps in the process, and the kinetics model which examines the temporal and spatial processes during one thermocycle. Most importantly, we conducted the first successful syntheses of a gene using small starting oligomers (tetramers). The synthesized sequence, 813 base pairs long, contained a 725 base pair gene, modified green fluorescent protein (mGFP), which has been shown to be a functional gene by cloning into cells and observing its green fluorescent product.

  17. Hole hopping rates in single strand oligonucleotides

    Energy Technology Data Exchange (ETDEWEB)

    Borrelli, Raffaele [Dipartimento di Scienze Agrarie, Forestali e Alimentari, Università di Torino, Largo Paolo Braccini 2, I-10095 Grugliasco, TO (Italy); Capobianco, Amedeo [Dipartimento di Chimica e Biologia, Università di Salerno, Via Giovanni Paolo II, I-84084 Fisciano, SA (Italy); Peluso, Andrea, E-mail: [Dipartimento di Chimica e Biologia, Università di Salerno, Via Giovanni Paolo II, I-84084 Fisciano, SA (Italy)


    Highlights: • DNA hole transfer rates have been computed. • Delocalized adenine domains significantly affect hole transfer rates in DNA. • Franck–Condon weighted density of state from DFT normal modes. • DNA application in molecular electronics. - Abstract: The rates of hole transfer between guanine and adenine in single strand DNA have been evaluated by using Fermi’s golden rule and Kubo’s generating function approach for the Franck–Condon weighted density of states. The whole sets of the normal modes and vibrational frequencies of the two nucleobases, obtained at DFT/B3LYP level of calculation, have been considered in computations. The results show that in single strand the pyramidalization/planarization mode of the amino groups of both nucleobases plays the major role. At room temperature, the Franck–Condon density of states extends over a wide range of hole site energy difference, 0–1 eV, giving some hints about the design of oligonucleotides of potential technological interest.

  18. Silver and Cyanine Staining of Oligonucleotides in Polyacrylamide Gel.

    Directory of Open Access Journals (Sweden)

    Weizhong Tang

    Full Text Available To explore why some oligonucleotides in denaturing polyacrylamide gel could not be silver-stained, 134 different oligonucleotides were analyzed using denaturing polyacrylamide gel electrophoresis stained with silver and asymmetric cyanine. As a result, we found that the sensitivity of oligos (dA, (dC, (dG and (dT to silver staining could be ranged as (dA > (dG > (dC > (dT from high to low. It was unexpected that oligo (dT was hard to be silver-stained. Moreover, the silver staining of an oligonucleotide containing base T could be partially or completely inhibited by base T. The inhibition of silver staining by base T was a competitive inhibition which could be affected by the amounts of the argyrophil nucleobase and base T, the cis-distance between the argyrophil nucleobase and base T, and the gel concentration. The changes of the intensity of an oligonucleotide band caused by the changes of DNA base composition were diverse and interesting. The intensity of some oligonucleotide bands would significantly change when the changes of DNA base composition accumulated to a certain extent (usually ≥ 4 nt. The sensitivity of cyanine staining of ≤ 11-nt long oligonucleotides could be enhanced about 250-fold by fixing the gels with methanol fixing solution.

  19. Brain Basics

    Medline Plus

    Full Text Available ... Basics will introduce you to some of this science, such as: How the brain develops How genes and the environment affect the brain The basic structure of the brain How different parts of the brain communicate and work with each other How changes in the brain ...

  20. The Team Boat Exercise: Enhancing Team Communication Midsemester (United States)

    Cox, Pamela L.; Friedman, Barry A.


    This paper discusses the Team Boat Exercise, which was developed to provide students with a mechanism for addressing team problems and enhancing team communication midsemester. The inspiration for the exercise came from a video by Prentice Hall, Inc. (2001). Part III of the video, entitled "Corporate Coaching," shows senior staff members from the…

  1. 77 FR 17084 - National Boating Safety Advisory Council (United States)


    ... Safety Strategic Planning Subcommittee will meet on Friday, April 13, 2012 from 1:30 p.m. to 3:45 p.m... comment. Saturday, April 14, 2012 (1) Receipt and Discussion of the Strategic Planning, Boats & Associated... meetings may conclude early if NBSAC has completed all business. All written materials, comments,...

  2. Carboranyl oligonucleotides. 3. Biochemical properties of oligonucleotides containing 5-(o-carboranyl-l-yl)-2{prime}-deoxyuridine

    Energy Technology Data Exchange (ETDEWEB)

    Lesnikowski, Z.J.; Fulcrand, G.; Lloyd, R.M. Jr. [Veterans Affairs Medical Center and Georgia Research Center for AIDS and HIV Infections, Decatur, GA (United States)]|[Emory Univ. School of Medicine, Atlanta, GA (United States)


    Boronated oligonucleotides are potential candidates for boron neutron capture therapy, antisense technology, and as tools in molecular biology. The biological properties of dodecathymidylic acids containing one or more 5-(o-carboran-l-yl)-2{prime}-deoxyuridine residues at different locations within the oligonucleotide chain were studied. 5-(o-carboran-l-yl)-2{prime}-deoxyuridine containing oligonucleotides manifested marked increased lipophilicity and resistance to 3{prime}- or 5{prime}-phosphodiesterases compared to the corresponding unmodified oligomer. They were substrates for T4 polynucleotide kinase and primers for Escherichia coli polymerase I and human immunodeficiency virus type 1 reverse transcriptase but not for human DNA polymerase {alpha} and {beta}. They also formed heteroduplexes that were substrates for E. coli RNase H, an essential property for antisense technology. These studies indicate that the carboranyl-containing oligonucleotides have desirable properties that need to be exploited further in the design of novel biopharmaceuticals. 33 refs., 2 figs., 1 tab.

  3. Targeting of single stranded oligonucleotides through metal-induced cyclization of short complementary strands : Targeting of single stranded oligonucleotides


    Freville, Fabrice; Richard, Tristan; Bathany, Katell; Moreau, Serge


    International audience; A new strategy to cyclize a short synthetic oligonucleotide on a DNA or a RNA target strand is described. This one relies on a metal-mediated cyclization of short synthetic oligonucleotides conjugated with two chelating 2,2':6',2”-terpyridine moieties at their 3' and 5' ends. Cyclization following metal addition (Zn2+, Fe2+) was demonstrated using UV monitored thermal denaturation experiments, mass spectrometry analysis and gel shift assays. NMR experiments were used t...

  4. Basic hydraulics

    CERN Document Server

    Smith, P D


    BASIC Hydraulics aims to help students both to become proficient in the BASIC programming language by actually using the language in an important field of engineering and to use computing as a means of mastering the subject of hydraulics. The book begins with a summary of the technique of computing in BASIC together with comments and listing of the main commands and statements. Subsequent chapters introduce the fundamental concepts and appropriate governing equations. Topics covered include principles of fluid mechanics; flow in pipes, pipe networks and open channels; hydraulic machinery;

  5. 78 FR 42733 - Safety Zone; Cleveland Dragon Boat Festival and Head of the Cuyahoga, Cuyahoga River, Cleveland, OH (United States)


    ... SECURITY Coast Guard 33 CFR Part 165 RIN 1625-AA00 Safety Zone; Cleveland Dragon Boat Festival and Head of... during the Dragon Boat Festival and Head of the Cuyahoga boating events. The safety zone established by... (HOTC) rowing regatta has occurred annually for over a decade and the Dragon Boat Festival for the...

  6. 77 FR 15600 - Special Local Regulation; Emerald Coast Super Boat Grand Prix; Saint Andrew Bay; Panama City, FL (United States)


    ... SECURITY Coast Guard 33 CFR Part 100 RIN 1625-AA08 Special Local Regulation; Emerald Coast Super Boat Grand... crews, vessels, and persons on navigable waters during the Emerald Coast Super Boat Grand Prix high... the Emerald Coast Super Boat Grand Prix high speed boat races. Basis and Purpose On January 31,...

  7. Brain Basics

    Medline Plus

    Full Text Available ... depression. The Growing Brain Inside the Brain: Neurons & Neural Circuits Neurons are the basic working unit of ... but sometimes give rise to disabilities or diseases. neural circuit —A network of neurons and their interconnections. ...

  8. Brain Basics

    Medline Plus

    Full Text Available ... Real Life Brain Basics in Real Life—How Depression affects the Brain Meet Sarah Sarah is a ... blues" from time to time. In contrast, major depression is a serious disorder that lasts for weeks. ...

  9. Schizophrenia Basics (United States)

    ... I know with schizophrenia? For More Information Share Schizophrenia Basics Download PDF Download ePub Order a free hardcopy What is schizophrenia? Schizophrenia is a serious mental disorder that affects ...

  10. Brain Basics

    Medline Plus

    Full Text Available ... News About Us Home > Health & Education > Educational Resources Brain Basics Introduction The Growing Brain The Working Brain ... to mental disorders, such as depression. The Growing Brain Inside the Brain: Neurons & Neural Circuits Neurons are ...

  11. Brain Basics (United States)

    ... News About Us Home > Health & Education > Educational Resources Brain Basics Introduction The Growing Brain The Working Brain ... to mental disorders, such as depression. The Growing Brain Inside the Brain: Neurons & Neural Circuits Neurons are ...

  12. Fluoridation Basics (United States)

    ... Page Basic Information About Fluoride Benefits: Strong Teeth History of Fluoride in Water Cost: Saves Money, Saves Teeth Fluoride in the Water Today The mineral fluoride occurs naturally on earth and is released from rocks into the soil, ...

  13. Basic Finance (United States)

    Vittek, J. F.


    A discussion of the basic measures of corporate financial strength, and the sources of the information is reported. Considered are: balance sheet, income statement, funds and cash flow, and financial ratios.

  14. Brain Basics

    Medline Plus

    Full Text Available ... science, such as: How the brain develops How genes and the environment affect the brain The basic ... that with brain development in people mental disorders. Genes and environmental cues both help to direct this ...

  15. Brain Basics

    Medline Plus

    Full Text Available ... in the anatomy, physiology, and chemistry of the nervous system. When the brain cannot effectively coordinate the billions ... the basic working unit of the brain and nervous system. These cells are highly specialized for the function ...

  16. Biominetic High Density Lipoproteins for the Delivery of Therapeutic Oligonucleotides (United States)

    Tripathy, Sushant

    Advances in nanotechnology have brought about novel inorganic and hybrid nanoparticles with unique physico-chemical properties that make them suitable for a broad range of applications---from nano-circuitry to drug delivery. A significant part of those advancements have led to ground-breaking discoveries that have changed the approaches to formulation of therapeutics against diseases, such as cancer. Now-a-days the focus does not lie solely on finding a candidate small-molecule therapeutic with minimal adverse effects, but researchers are looking up to nanoparticles to improve biodistribution and biocompatibility profile of clinically proven therapeutics. The plethora of conjugation chemistries offered by currently extant inorganic nanoparticles have, in recent years, led to great leaps in the field of biomimicry---a modality that promises high biocompatibility. Further, in the pursuit of highly specific therapeutic molecules, researchers have turned to silencing oligonucleotides and some have already brought together the strengths of nanoparticles and silencing oligonucleotides in search of an efficacious therapy for cancer with minimal adverse effects. This dissertation work focuses on such a biomimetic platform---a gold nanoparticle based high density lipoprotein biomimetic (HDL NP), for the delivery of therapeutic oligonucleotides. The first chapter of this body of work introduces the molecular target of the silencing oligonucleotides---VEGFR2, and its role in the progression of solid tumor cancers. The background information also covers important aspects of natural high density lipoproteins (HDL), especially their innate capacity to bind and deliver exogenous and endogenous silencing oligonucleotides to tissues that express their high affinity receptor SRB1. We subsequently describe the synthesis of the biomimetic HDL NP and its oligonucleotide conjugates, and establish their biocompatibility. Further on, experimental data demonstrate the efficacy of silencing

  17. China's export of boats valued at over200million USD in2010%China's export of boats valued at over 200 million USD in 2010

    Institute of Scientific and Technical Information of China (English)

    Zhang Shouchun


    @@ In 2010, China's economy saw a relatively good recovery.The economic operation of the shipbuilding industry was better than expected and the import and export trade of ships developed at high speeds.According to the statistics of China's custom house, thetotal value of the import and export of boats was 295 million USD, up 40.75% from a year ago; the total quantity in import and export trade was 2057200 boats, up 15.26% in quantity from a year ago. Last year, the value of China's export of boats was 204 million USD, up 19.64%;2028700 boats were exported, up 14.79% in the quantity of boats from a year ago.The ratio of the export of boats to the export of ships in 2010 was 1%; the increase rate of boat export was much lower than that of ship export, which was 42.7%.In the first half of 2010, the export trade of boats showed a highspeed increase with monthly average export value of 14.655 million USD while in August and September, export decreased at a significant rate with the lowest value of 9.7324 million USD in September.

  18. Basic electronics

    CERN Document Server

    Holbrook, Harold D


    Basic Electronics is an elementary text designed for basic instruction in electricity and electronics. It gives emphasis on electronic emission and the vacuum tube and shows transistor circuits in parallel with electron tube circuits. This book also demonstrates how the transistor merely replaces the tube, with proper change of circuit constants as required. Many problems are presented at the end of each chapter. This book is comprised of 17 chapters and opens with an overview of electron theory, followed by a discussion on resistance, inductance, and capacitance, along with their effects on t

  19. Impact of boat-generated waves on intertidal estuarine sediments (United States)

    Blanpain, O.; Deloffre, J.; Lafite, R.; Gomit, G.; Calluaud, D.; David, L.


    Hydrodynamics in the macrotidal Seine estuary (France) are controlled by the semi-diurnal tidal regime modulated seasonally by the fluvial discharge. Wind effect on sediment transport (through wind waves and swell) is observed at the mouth of the estuary. Over the last century, authorities have put emphasis on facilitating economic exchanges by means of embankment building and increased dredging activity. These developments led to allow and secure sea vessel traffic in the Seine estuary (from its mouth to the port of Rouen, 125 km upstream) but they also resulted in a change of estuarine hydrodynamics and sediment transport features. A riversides restoration policy has been recently started by port authorities. In this context, the objective of the field-based study presented is to connect vessel characteristics (i.e. speed, draft...), boat-generated waves and their sedimentary impacts. Such information will be used by stakeholders to manage riverside. The natural intertidal site of interest is located in the fluvial freshwater part of the Seine estuary characterized by a 4.5 m maximum tidal range. The foreshore slope is gently decreasing and surface sediments are composed of fine to coarse sand with occasional mud drapes. In order to decipher boat-generated events, the sampling strategy is based on continuous ADV measurements coupled with a turbidimeter and an altimeter to study sediment dynamics. These instruments are settled in the lower part of the foreshore (i) to obtain a significant dataset (i.e. oceanic instruments are not measuring in air) on a zone statically affected by boat waves and (ii) because most of boat traffic occurs during early flood or late ebb period. Spatial variations are assessed along a cross-section through grain-size analysis of surface sediments and topography measurements using pole technique. Results enhance hydrodynamic and sedimentary impacts of boat-generated waves compared respectively to tidal and wind effects. Long

  20. Solar Photovoltaic Powered Sailing Boat Using Buck Converter

    Directory of Open Access Journals (Sweden)

    Soumya Das


    Full Text Available The main objective of this paper is to establish technical and economical aspects of the application of stand-alone photovoltaic (PV system in sailing boat using a buck converter in order to enhance the power generation and also to minimize the cost. Performance and control of dc-dc converter, suitable for photovoltaic (PV applications, is presented here. A buck converter is employed here which extracts complete power from the PV source and feeds into the dc load. The power, which is fed into the load, is sufficient to drive a boat . With the help of matlab simulink software PV module and buck model has been designed and simulated and also compared with theoretical predictions.

  1. [Recreational boating accidents--Part 1: Catamnestic study]. (United States)

    Lignitz, Eberhard; Lustig, Martina; Scheibe, Ernst


    Deaths on the water are common in the autopsy material of medicolegal institutes situated on the coast or big rivers and lakes (illustrated by the example of the Institute of Legal Medicine of Greifswald University). They mostly occur during recreational boating activities. Apart from hydro-meteorological influences, human error is the main cause of accidents. Often it is not sufficiently kept in mind whether the boat crew is fit for sailing and proper seamanship is ensured. Drowning (following initial hypothermia) is the most frequent cause of death. Medicolegal aspects are not decisive for ordering a forensic autopsy. As statistics are not compiled in a uniform way, a comparison of the data of different institutions engaged in investigating deaths at sea and during water sports activities is hardly possible, neither on a national nor an international basis--and the reconstruction of aquatic accidents is generally difficult. Fatal accidents can only be prevented by completely clarifying their causes.

  2. Investigations of oligonucleotide usage variance within and between prokaryotes

    DEFF Research Database (Denmark)

    Bohlin, J.; Skjerve, E.; Ussery, David


    Oligonucleotide usage in archaeal and bacterial genomes can be linked to a number of properties, including codon usage (trinucleotides), DNA base-stacking energy (dinucleotides), and DNA structural conformation (di-to tetranucleotides). We wanted to assess the statistical information potential...... was that prokaryotic chromosomes can be described by hexanucleotide frequencies, suggesting that prokaryotic DNA is predominantly short range correlated, i. e., information in prokaryotic genomes is encoded in short oligonucleotides. Oligonucleotide usage varied more within AT-rich and host-associated genomes than...... in GC-rich and free-living genomes, and this variation was mainly located in non-coding regions. Bias (selectional pressure) in tetranucleotide usage correlated with GC content, and coding regions were more biased than non-coding regions. Non-coding regions were also found to be approximately 5.5% more...

  3. Delivery of RNAi-Based Oligonucleotides by Electropermeabilization

    Directory of Open Access Journals (Sweden)

    Muriel Golzio


    Full Text Available For more than a decade, understanding of RNA interference (RNAi has been a growing field of interest. The potent gene silencing ability that small oligonucleotides have offers new perspectives for cancer therapeutics. One of the present limits is that many biological barriers exist for their efficient delivery into target cells or tissues. Electropermeabilization (EP is one of the physical methods successfully used to transfer small oligonucleotides into cells or tissues. EP consists in the direct application of calibrated electric pulses to cells or tissues that transiently permeabilize the plasma membranes, allowing efficient in vitro and in vivo. cytoplasmic delivery of exogenous molecules. The present review reports on the type of therapeutic RNAi-based oligonucleotides that can be electrotransferred, the mechanism(s of their electrotransfer and the technical settings for pre-clinical purposes.

  4. Rapid and accurate synthesis of TALE genes from synthetic oligonucleotides. (United States)

    Wang, Fenghua; Zhang, Hefei; Gao, Jingxia; Chen, Fengjiao; Chen, Sijie; Zhang, Cuizhen; Peng, Gang


    Custom synthesis of transcription activator-like effector (TALE) genes has relied upon plasmid libraries of pre-fabricated TALE-repeat monomers or oligomers. Here we describe a novel synthesis method that directly incorporates annealed synthetic oligonucleotides into the TALE-repeat units. Our approach utilizes iterative sets of oligonucleotides and a translational frame check strategy to ensure the high efficiency and accuracy of TALE-gene synthesis. TALE arrays of more than 20 repeats can be constructed, and the majority of the synthesized constructs have perfect sequences. In addition, this novel oligonucleotide-based method can readily accommodate design changes to the TALE repeats. We demonstrated an increased gene targeting efficiency against a genomic site containing a potentially methylated cytosine by incorporating non-conventional repeat variable di-residue (RVD) sequences.

  5. Deployment of Lightweight Shock Mitigating Boat Manufacturing Innovation (United States)


    VARTM Hull 1267 760 Deck 702 399 Console 540 350 Total 2509 1509 % Current 100% 60% % Savings - 40% Deployment of Lightweight Shock Mitigating...1430 47 Weight Evaluation- Baseline Boat Weights (lb) Segment Baseline 850 A 850 B 850 B-OM Fabrication Method Open Mold Open Mold VARTM Open...Segment Baseline 850 A 850 B 850 B-OM Fabrication Method Open Mold Open Mold VARTM Open Mold Hull 1115 843 717 789 Deck 665 587 481 529

  6. U-Boat Worx出租私人用潜艇

    Institute of Scientific and Technical Information of China (English)


    荷兰潜艇制造商U—Boat Worx(UBW)现推出两款潜艇对外出租,使个人和科学家们能够深入探索神秘的海底世界。客户可自行探索或专门欣赏海底世界的美景与水生物。

  7. Investigation of longitudinal control system for a small hydrofoil boat (United States)

    Phillips, W. H.; Shaughnessy, J. D.


    An analysis of a hydromechanical system for longitudinal control of a small hydrofoil boat is presented. The system incorporates height and acceleration sensors operating flaps on the foils through a mechanical linkage. Effects of some of the system parameters on the stability and response to waves are shown. The results indicate that the system is capable of providing adequate stability, but the response to stern waves at low frequencies is larger than desired.

  8. Oligonucleotide Therapy for Obstructive and Restrictive Respiratory Diseases. (United States)

    Liao, Wupeng; Dong, Jinrui; Peh, Hong Yong; Tan, Lay Hong; Lim, Kah Suan; Li, Li; Wong, Wai-Shiu Fred


    Inhaled oligonucleotide is an emerging therapeutic modality for various common respiratory diseases, including obstructive airway diseases like asthma and chronic obstructive pulmonary disease (COPD) and restrictive airway diseases like idiopathic pulmonary fibrosis (IPF). The advantage of direct accessibility for oligonucleotide molecules to the lung target sites, bypassing systemic administration, makes this therapeutic approach promising with minimized potential systemic side effects. Asthma, COPD, and IPF are common chronic respiratory diseases, characterized by persistent airway inflammation and dysregulated tissue repair and remodeling, although each individual disease has its unique etiology. Corticosteroids have been widely prescribed for the treatment of asthma, COPD, and IPF. However, the effectiveness of corticosteroids as an anti-inflammatory drug is limited by steroid resistance in severe asthma, the majority of COPD cases, and pulmonary fibrosis. There is an urgent medical need to develop target-specific drugs for the treatment of these respiratory conditions. Oligonucleotide therapies, including antisense oligonucleotide (ASO), small interfering RNA (siRNA), and microRNA (miRNA) are now being evaluated both pre-clinically and clinically as potential therapeutics. The mechanisms of action of ASO and siRNA are highly target mRNA specific, ultimately leading to target protein knockdown. miRNA has both biomarker and therapeutic values, and its knockdown by a miRNA antagonist (antagomir) has a broader but potentially more non-specific biological outcome. This review will compile the current findings of oligonucleotide therapeutic targets, verified in various respiratory disease models and in clinical trials, and evaluate different chemical modification approaches to improve the stability and potency of oligonucleotides for the treatment of respiratory diseases.

  9. Oligonucleotide Therapy for Obstructive and Restrictive Respiratory Diseases

    Directory of Open Access Journals (Sweden)

    Wupeng Liao


    Full Text Available Inhaled oligonucleotide is an emerging therapeutic modality for various common respiratory diseases, including obstructive airway diseases like asthma and chronic obstructive pulmonary disease (COPD and restrictive airway diseases like idiopathic pulmonary fibrosis (IPF. The advantage of direct accessibility for oligonucleotide molecules to the lung target sites, bypassing systemic administration, makes this therapeutic approach promising with minimized potential systemic side effects. Asthma, COPD, and IPF are common chronic respiratory diseases, characterized by persistent airway inflammation and dysregulated tissue repair and remodeling, although each individual disease has its unique etiology. Corticosteroids have been widely prescribed for the treatment of asthma, COPD, and IPF. However, the effectiveness of corticosteroids as an anti-inflammatory drug is limited by steroid resistance in severe asthma, the majority of COPD cases, and pulmonary fibrosis. There is an urgent medical need to develop target-specific drugs for the treatment of these respiratory conditions. Oligonucleotide therapies, including antisense oligonucleotide (ASO, small interfering RNA (siRNA, and microRNA (miRNA are now being evaluated both pre-clinically and clinically as potential therapeutics. The mechanisms of action of ASO and siRNA are highly target mRNA specific, ultimately leading to target protein knockdown. miRNA has both biomarker and therapeutic values, and its knockdown by a miRNA antagonist (antagomir has a broader but potentially more non-specific biological outcome. This review will compile the current findings of oligonucleotide therapeutic targets, verified in various respiratory disease models and in clinical trials, and evaluate different chemical modification approaches to improve the stability and potency of oligonucleotides for the treatment of respiratory diseases.

  10. A novel catechol-based universal support for oligonucleotide synthesis. (United States)

    Anderson, Keith M; Jaquinod, Laurent; Jensen, Michael A; Ngo, Nam; Davis, Ronald W


    A novel universal support for deoxyribo- and ribonucleic acid synthesis has been developed. The support, constructed from 1,4-dimethoxycatechol, represents an improvement over existing universal supports because of its ability to cleave and deprotect under mild conditions in standard reagents. Because no nonvolatile additives are required for cleavage and deprotection, the synthesized oligonucleotides do not require purification prior to use in biochemical assays. Using reverse phase HPLC and electrospray mass spectroscopy, it was determined that oligonucleotides synthesized on the universal support (UL1) 3'-dephosphorylate quickly (9 h in 28-30% ammonium hydroxide (NH4OH) at 55 degrees C, 2 h in 28-30% NH4OH at 80 degrees C, or <1 h in ammonium hydroxide/methylamine (1:1) (AMA) at 80 degrees C). Oligonucleotides used as primers for the polymerase chain reaction (PCR) assay were found to perform identically to control primers, demonstrating full biological compatibility. In addition, a method was developed for sintering the universal support directly into a filter plug which can be pressure fit into the synthesis column of a commercial synthesizer. The universal support plugs allow the synthesis of high-quality oligonucleotides at least 120 nucleotides in length, with purity comparable to non-universal commercial supports and approximately 50% lower reagent consumption. The universal support plugs are routinely used to synthesize deoxyribo-, ribo-, 3'-modified, 5'-modified, and thioated oligonucleotides. The flexibility of the universal support and the efficiency of 3'-dephosphorylation are expected to increase the use of universal supports in oligonucleotide synthesis.

  11. Snohomish Estuary Wetlands Study. Volume II. Basic Information and Evaluation (United States)


    moorage and may be used by boat,ýrs or swimmers . A boom is basically a floating log, moored a:t each end, the purpose o’ which is to enclose an area of...ndpbl ting or environment In which thle imt . humain safety, health and welfart’:; and 239.14 Application of XO to Civil Works ural functions of the

  12. Ethanol Basics

    Energy Technology Data Exchange (ETDEWEB)



    Ethanol is a widely-used, domestically-produced renewable fuel made from corn and other plant materials. More than 96% of gasoline sold in the United States contains ethanol. Learn more about this alternative fuel in the Ethanol Basics Fact Sheet, produced by the U.S. Department of Energy's Clean Cities program.

  13. Body Basics (United States)

    ... more about how the body works, what basic human anatomy is, and what happens when parts of the body don't function properly. Blood Bones, Muscles, and Joints Brain and Nervous System Digestive System Endocrine System Eyes Female Reproductive System Heart and Circulatory System Immune ...

  14. Brain Basics

    Medline Plus

    Full Text Available ... such as depression. The Growing Brain Inside the Brain: Neurons & Neural Circuits Neurons are the basic working unit ... final destination. Chemical signals from other cells guide neurons in forming various brain structures. Neighboring neurons make connections with each other ...

  15. Insulin Basics (United States)

    ... Honor Become a Member En Español Type 1 Type 2 About Us Online Community Meal Planning Sign In Search: Search More Sites Search ≡ Are You At Risk? Diabetes Basics Living with Diabetes Food & Fitness In My ... Diabetes and Learning About Prediabetes Type 2 Diabetes Risk Test Lower Your Risk Healthy ...

  16. Synthesis of Peptide-Oligonucleotide Conjugates Using a Heterobifunctional Crosslinker (United States)

    Williams, Berea A.R.; Chaput, John C.


    Peptide-oligonucleotide conjugates (POCs) are molecular chimeras composed of a nucleic acid moiety covalently attached to a polypeptide moiety. POCs have been used in numerous applications from therapeutics to nanotechnology, and most recently as combinatorial agents in the assembly of bivalent protein affinity reagents. This unit describes the synthesis and purification of POC molecules using the heterobifunctional crosslinking reagent succinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC), which enables amine-modified oligonucleotides to become covalently linked to cysteine-modified polypeptides. This solution-based protocol consists of a two-step synthesis followed by a single purification step. PMID:20827717

  17. Chemical phosphorylation of deoxyribonucleosides and thermolytic DNA oligonucleotides. (United States)

    Ausín, Cristina; Grajkowski, Andrzej; Cieślak, Jacek; Beaucage, Serge L


    The phosphorylating reagent bis[S-(4,4'-dimethoxytrityl)-2-mercaptoethyl]-N,N-diisopropylphosphoramidite is prepared in three steps from commercial methyl thioglycolate and diisopropylphosphoramidous dichloride. The phosphorylating reagent has been used successfully in the solid-phase synthesis of deoxyribonucleoside 5'-/3'-phosphate or -thiophosphate monoesters and oligonucleotide 5'-phosphate/-thiophosphate monoesters. Bis[S-(4,4'-dimethoxytrityl)-2-mercaptoethyl]-N,N-diisopropylphosphoramidite has also been employed in the construction of a thermolytic dinucleotide prodrug model to evaluate the ability of the reagent to produce thermosentive oligonucleotide prodrugs under mild temperature conditions ( approximately 25 degrees C) for potential therapeutic applications.

  18. Versatile functionalization of nanoelectrodes by oligonucleotides via pyrrole electrochemistry. (United States)

    Descamps, Emeline; Nguyen, Khoa; Bouchain-Gautier, Christelle; Filoramo, Arianna; Goux-Capes, Laurence; Goffman, Marcello; Bourgoin, Jean-Philippe; Mailley, Pascal; Livache, Thierry


    Surface modification at the nanometer scale is a challenge for the future of molecular electronics. In particular, the precise anchoring and electrical addressing of biological scaffolds such as complex DNA nanonetworks is of importance for generating bio-directed assemblies of nano-objects for nanocircuit purposes. Herein, we consider the individual modification of nanoelectrodes with different oligonucleotide sequences by an electrochemically driven co-polymerization process of pyrrole and modified oligonucleotide sequences bearing pyrrole monomers. We demonstrate that this one-step technique presents the advantages of simplicity, localization of surface modification, mechanical, biological and chemical stability of the coatings, and high lateral resolution.

  19. Wavelet basics

    CERN Document Server

    Chan, Y T


    Since the study of wavelets is a relatively new area, much of the research coming from mathematicians, most of the literature uses terminology, concepts and proofs that may, at times, be difficult and intimidating for the engineer. Wavelet Basics has therefore been written as an introductory book for scientists and engineers. The mathematical presentation has been kept simple, the concepts being presented in elaborate detail in a terminology that engineers will find familiar. Difficult ideas are illustrated with examples which will also aid in the development of an intuitive insight. Chapter 1 reviews the basics of signal transformation and discusses the concepts of duals and frames. Chapter 2 introduces the wavelet transform, contrasts it with the short-time Fourier transform and clarifies the names of the different types of wavelet transforms. Chapter 3 links multiresolution analysis, orthonormal wavelets and the design of digital filters. Chapter 4 gives a tour d'horizon of topics of current interest: wave...

  20. Oligonucleotide-directed mutagenesis for precision gene editing. (United States)

    Sauer, Noel J; Mozoruk, Jerry; Miller, Ryan B; Warburg, Zachary J; Walker, Keith A; Beetham, Peter R; Schöpke, Christian R; Gocal, Greg F W


    Differences in gene sequences, many of which are single nucleotide polymorphisms, underlie some of the most important traits in plants. With humanity facing significant challenges to increase global agricultural productivity, there is an urgent need to accelerate the development of these traits in plants. oligonucleotide-directed mutagenesis (ODM), one of the many tools of Cibus' Rapid Trait Development System (RTDS(™) ) technology, offers a rapid, precise and non-transgenic breeding alternative for trait improvement in agriculture to address this urgent need. This review explores the application of ODM as a precision genome editing technology, with emphasis on using oligonucleotides to make targeted edits in plasmid, episomal and chromosomal DNA of bacterial, fungal, mammalian and plant systems. The process of employing ODM by way of RTDS technology has been improved in many ways by utilizing a fluorescence conversion system wherein a blue fluorescent protein (BFP) can be changed to a green fluorescent protein (GFP) by editing a single nucleotide of the BFP gene (CAC→TAC; H66 to Y66). For example, dependent on oligonucleotide length, applying oligonucleotide-mediated technology to target the BFP transgene in Arabidopsis thaliana protoplasts resulted in up to 0.05% precisely edited GFP loci. Here, the development of traits in commercially relevant plant varieties to improve crop performance by genome editing technologies such as ODM, and by extension RTDS, is reviewed.

  1. Lipid-modified G4-decoy oligonucleotide anchored to nanoparticles

    DEFF Research Database (Denmark)

    Cogoi, S; Jakobsen, U; Pedersen, E B


    KRAS is mutated in >90% of pancreatic ductal adenocarcinomas. As its inactivation leads to tumour regression, mutant KRAS is considered an attractive target for anticancer drugs. In this study we report a new delivery strategy for a G4-decoy oligonucleotide that sequesters MAZ, a transcription fa...

  2. Regioselective immobilization of short oligonucleotides to acrylic copolymer gels. (United States)

    Timofeev, E; Kochetkova, S V; Mirzabekov, A D; Florentiev, V L


    Four types of polyacrylamide or polydimethyl-acrylamide gels for regioselective (by immobilization at the 3' end) of short oligonucleotides have been designed for use in manufacturing oligonucleotide microchips. Two of these supports contain amino or aldehyde groups in the gel, allowing coupling with oligonucleotides bearing aldehyde or amino groups, respectively, in the presence of a reducing agent. The aldehyde gel support showed a higher immobilization efficiency relative to the amino gel. Of all reducing agents tested, the best results were obtained with a pyridine-borane complex. The other supports are based on an acrylamide gel activated with glutaraldehyde or a hydroxyalkyl-functionalized gel treated with mesyl chloride. The use of dimethylacrylamide instead of acrylamide allows subsequent gel modifications in organic solvents. All the immobilization methods are easy and simple to perform, give high and reproducible yields, allow long durations of storage of the activated support, and provide high stability of attachment and low non-specific binding. Although these gel supports have been developed for preparing oligonucleotide microchips, they may be used for other purposes as well. PMID:8774893

  3. LNA 5'-phosphoramidites for 5'→3'-oligonucleotide synthesis

    DEFF Research Database (Denmark)

    Madsen, Andreas Stahl; Kumar, Santhosh T.; Wengel, Jesper


    Hereby we report an efficient synthesis of LNA thymine and LNA 5-methylcytosine 5′-phosphoramidites, allowing incorporation of LNA thymine and LNA 5-methylcytosine into oligonucleotides synthesized in the 5′→3′ direction. Key steps include regioselective enzymatic benzoylation of the 5′-hydroxy g...

  4. Systematic design of mouse Vh gene family-specific oligonucleotides

    NARCIS (Netherlands)

    Seijen, AM; Seijen, HG; Bos, NA


    Kabat's database has often been used to design mouse Vh gene-specific 5 ' primers. The emphasis was mostly on constructing a universal (degenerate) 5 ' primer or 5 ' primer set, which would be able to match every mouse Vh gene. We were interested in finding oligonucleotides that could be used as pri

  5. Chromosome-specific painting in Cucumis species using bulked oligonucleotides (United States)

    Chromosome-specific painting is a powerful technique in molecular cytogenetic and genome research. We developed an oligonucleotide (oligo)-based chromosome painting technique in cucumber (Cucumis sativus) that will be applicable in any plant species with a sequenced genome. Oligos specific to a sing...

  6. Oligonucleotides with 1,4-dioxane-based nucleotide monomers

    DEFF Research Database (Denmark)

    Madsen, Andreas S; Wengel, Jesper


    An epimeric mixture of H-phosphonates 5R and 5S has been synthesized in three steps from known secouridine 1. Separation of the epimers has been accomplished by RP-HPLC, allowing full characterization and incorporation of monomers X and Y into 9-mer oligonucleotides using H-phosphonates building ...

  7. The behavioural response of Australian fur seals to motor boat noise.

    Directory of Open Access Journals (Sweden)

    Joy S Tripovich

    Full Text Available Australian fur seals breed on thirteen islands located in the Bass Strait, Australia. Land access to these islands is restricted, minimising human presence but boat access is still permissible with limitations on approach distances. Thirty-two controlled noise exposure experiments were conducted on breeding Australian fur seals to determine their behavioural response to controlled in-air motor boat noise on Kanowna Island (39°10'S, 146°18'E. Our results show there were significant differences in the seals' behaviour at low (64-70 dB versus high (75-85 dB sound levels, with seals orientating themselves towards or physically moving away from the louder boat noise at three different sound levels. Furthermore, seals responded more aggressively with one another and were more alert when they heard louder boat noise. Australian fur seals demonstrated plasticity in their vocal responses to boat noise with calls being significantly different between the various sound intensities and barks tending to get faster as the boat noise got louder. These results suggest that Australian fur seals on Kanowna Island show behavioural disturbance to high level boat noise. Consequently, it is recommended that an appropriate level of received boat sound emissions at breeding fur seal colonies be below 74 dB and that these findings be taken into account when evaluating appropriate approach distances and speed limits for boats.

  8. The Behavioural Response of Australian Fur Seals to Motor Boat Noise (United States)

    Tripovich, Joy S.; Hall-Aspland, Sophie; Charrier, Isabelle; Arnould, John P. Y.


    Australian fur seals breed on thirteen islands located in the Bass Strait, Australia. Land access to these islands is restricted, minimising human presence but boat access is still permissible with limitations on approach distances. Thirty-two controlled noise exposure experiments were conducted on breeding Australian fur seals to determine their behavioural response to controlled in-air motor boat noise on Kanowna Island (39°10′S, 146°18′E). Our results show there were significant differences in the seals' behaviour at low (64–70 dB) versus high (75–85 dB) sound levels, with seals orientating themselves towards or physically moving away from the louder boat noise at three different sound levels. Furthermore, seals responded more aggressively with one another and were more alert when they heard louder boat noise. Australian fur seals demonstrated plasticity in their vocal responses to boat noise with calls being significantly different between the various sound intensities and barks tending to get faster as the boat noise got louder. These results suggest that Australian fur seals on Kanowna Island show behavioural disturbance to high level boat noise. Consequently, it is recommended that an appropriate level of received boat sound emissions at breeding fur seal colonies be below 74 dB and that these findings be taken into account when evaluating appropriate approach distances and speed limits for boats. PMID:22623998

  9. Dating the Bibong-ri Neolithic site in Korea: Excavating the oldest ancient boat

    Energy Technology Data Exchange (ETDEWEB)

    Park, Gyujun, E-mail: [Geochemical Analysis Center, Geologic Environment Division, Korea Institute of Geoscience and Mineral Resources 305-350 (Korea, Republic of); Kim, Jong-Chan [Department of Physics and Astronomy, Seoul National University, Seoul 151-742 (Korea, Republic of); Youn, Minyoung; Yun, Chongcheol; Kang, Jin; Song, Yong-Mi; Song, Su-Jin; Noh, Hye-Jin [Electrostatic Accelerator Research Center, NCIRF, Seoul National University, Seoul 151-742 (Korea, Republic of); Kim, Do-Kyun [Department of Physics and Astronomy, Seoul National University, Seoul 151-742 (Korea, Republic of); Im, Hack-Jong [Gimhae National Museum, Gimhae, Gyeongsangnam-do 621-060 (Korea, Republic of)


    The remains of an ancient wooden boat were unearthed at the Bibong-ri shell mound site. The site was located at Bibong-ri, Bugog-myeon, Changnyeong-gun, Gyeongsangnam-do in South Korea. A substantial fragment of the vessel was discovered in the lowest layer of the site. We collected 17 samples of charcoal and wood from pebble, sand, and shell layers. Sample preparation extracted the carbon from each sample material and converted it into graphite for AMS radiocarbon dating. Radiocarbon dates of the samples indicate that they belong to the Neolithic period and that the boat dates from ca. 5700 BC. To this point, the oldest known boat in the world has been a wooden boat dating from ca. 5500 BC in China. Other ancient boats from around the world include a logboat dating from ca. 3600 BC in Japan and a fleet of wooden boats dating from ca. 3000 BC in Egypt. The Bibong-ri boat is the first boat from the Neolithic period ever found in South Korea and must represent one of the world's oldest known boats.

  10. China Xiamen International Boat Show 2011——Making Yachting a new marine leisure fashion

    Institute of Scientific and Technical Information of China (English)


    The China (Xiamen) International Boat Show 2011 will be staged from November 4-7.Covering an area of 20,000 square meters with 300 berths,it will be China's largest boat show and an extravaganza for yacht manufacturers and enthusiasts.A series of other activities will be held alongside the boat show,such as a welcoming banquet,cruises,a seminar,a photography competition and a wedding upon one of the sailing vessels.The annual international boat show has become a unique attraction of southeast China's Xiamen City.

  11. Effects of fluid flow on the oligonucleotide folding in single-walled carbon nanotubes. (United States)

    Lim, M C G; Zhong, Z W


    This paper presents molecular-dynamics (MD) simulations of DNA oligonucleotide and water molecules translocating through carbon nanotube (CNT) channels. An induced pressure difference is applied to the system by pushing a layer of water molecules toward the flow direction to drive the oligonucleotide and other molecules. This MD simulation investigates the changes that occur in the conformation of the oligonucleotide due to water molecules in nanochannels while controlling the temperature and volume of the system in a canonical ensemble. The results show that the oligonucleotide in the (8,8)-(12,12) CNT channel forms a folded state at a lower pressure, whereas the oligonucleotide in the (10,10)-(14,14) CNT channel forms a folded state at a higher pressure instead. The van der Waals forces between the water molecules and the oligonucleotide suggest that the attraction between these two types of molecules results in the linear arrangements of the bases of the oligonucleotide. For a larger nanotube channel, the folding of the oligonucleotide is mainly dependent on the solvent (water molecules), whereas pressure, the size of the nanotube junction, and water molecules are the considering factors of the folding of the oligonucleotide at a smaller nanotube channel. For a folded oligonucleotide, the water distribution around the oligonucleotide is concentrated at a smaller range than that for the distribution around an unfolded oligonucleotide.

  12. Glycoclusters on oligonucleotide and PNA scaffolds: synthesis and applications. (United States)

    Spinelli, Nicolas; Defrancq, Eric; Morvan, François


    Conjugation of oligonucleotides (ONs) to a variety of reporter groups has been the subject of intensive research during the last decade. Conjugation is indeed of great interest because it can be used not only to improve the existing ONs properties but also to impart new ones. In this context tremendous efforts have been made to conjugate carbohydrate moieties to ONs. Indeed carbohydrates play an important role in biological processes such as signal transduction and cell adhesion through the recognition with sugar-binding proteins (i.e. lectins) located on the surface of cells. For this reason, carbohydrate-oligonucleotide conjugates (COCs) have been first developed for improving the poor cellular uptake or tissue specific delivery of ONs through receptor-mediated endocytosis. Besides the targeted ONs delivery, carbohydrate-oligonucleotide conjugates (COCs) are also evaluated in the context of carbohydrate biochips in which surface coating with carbohydrates is achieved by using the DNA-directed immobilization strategy (DDI). Peptide nucleic acids (PNAs) have also been extensively investigated as a surrogate of DNA for diverse applications. Therefore attachment of carbohydrate moieties to this class of molecules has been studied. The aforementioned applications of COCs require mimicking of the natural processes, in which the weak individual protein-carbohydrate binding is overcome by using multivalent interactions. This tutorial review focuses on the recent advances in carbohydrate-oligonucleotide conjugates and describes the major synthetic approaches available. In addition, an overview of applications that have been developed using various scaffolds allowing multivalent interactions is provided. Finally recent results on the use of peptide nucleic acids as oligonucleotides surrogate are described.

  13. Basic electronics

    CERN Document Server

    Tayal, DC


    The second edition of this book incorporates the comments and suggestions of my friends and students who have critically studied the first edition. In this edition the changes and additions have been made and subject matter has been rearranged at some places. The purpose of this text is to provide a comprehensive and up-to-date study of the principles of operation of solid state devices, their basic circuits and application of these circuits to various electronic systems, so that it can serve as a standard text not only for universities and colleges but also for technical institutes. This book

  14. Regression Basics

    CERN Document Server

    Kahane, Leo H


    Using a friendly, nontechnical approach, the Second Edition of Regression Basics introduces readers to the fundamentals of regression. Accessible to anyone with an introductory statistics background, this book builds from a simple two-variable model to a model of greater complexity. Author Leo H. Kahane weaves four engaging examples throughout the text to illustrate not only the techniques of regression but also how this empirical tool can be applied in creative ways to consider a broad array of topics. New to the Second Edition Offers greater coverage of simple panel-data estimation:

  15. Physiological and physical characteristics of elite dragon boat paddlers. (United States)

    Ho, Sarah R; Smith, Richard M; Chapman, Philip G; Sinclair, Peter J; Funato, Kazuo


    The objectives of this study were to profile the physiological and physical characteristics of elite dragon boat paddlers, to identify characteristics that predict race performance and to quantify the metabolic energy contributions to simulated 200-m and 500-m dragon boat racing. Eleven, national level, male, Japanese dragon boat paddlers completed a battery of tests on a paddling ergometer including an incremental maximal aerobic capacity test, a 2-minute maximal accumulated oxygen deficit (MAOD) test, and simulated 200-m and 500-m races. A physiological and physical profile of subjects was compiled. Results showed that 200-m race performance correlated with flexed arm girth and excess postexercise oxygen consumption (EPOC) measured in the 30 minutes after the MAOD test, whereas 500-m race performance correlated with body fat percentage, relaxed and flexed arm girth, MAOD, EPOC, and peak power during the MAOD test. Stepwise multiple regression revealed that flexed arm girth was the most powerful predictor of 200-m and 500-m race performance, followed by EPOC with the combination of these 2 factors able to explain 74% and 68% of the variance in 200-m and 500-m race performance, respectively. Aerobic energy contributions for 200-m (50 seconds) and 500-m (1 minute 50 seconds) races were (mean (95% confidence intervals)) 52.1% (range, 47.4-56.8%) and 67.5% (range, 60.1-77.8%), respectively. In conclusion, coaches should develop training programs targeted at developing upper-body musculature and increasing anaerobic capacity because these factors are the strongest predictors of 200-m and 500-m race performance. Given the substantial aerobic energy contributions even for a 200-m race event, coaches should aim to increase the maximal aerobic capacity of the paddler in preparation for both 200-m and 500-m events.

  16. On the Writing Style of The Open Boat

    Institute of Scientific and Technical Information of China (English)



      The Open Boat is one of the best realistic stories written by Stephen Crane, an early American naturalist and impressionist writer who dramatically presents the subtle relation between man and nature as well as that of man and society. His personal experience on the sea has an enormous impact on the story. This paper tries to carry out a detailed examination of the story in the light of writing style in order to explore his unique style and the beauty of the story.

  17. Compression fractures of the vertebrae during a "bumpy" boat ride.

    LENUS (Irish Health Repository)

    Chukwunyerenwa, C K


    INTRODUCTION: Compression fracture of the vertebrae is common, often the result of falls from height and motor vehicle accidents in the younger age groups. It can occur following minor trauma in the elderly and in those with osteoporosis. MATERIALS AND METHODS: We present an interesting case of compression fracture of the vertebral bodies occurring simultaneously in a couple during a boat ride while on holiday. One individual had fracture of the T8, while the other fractured the L1 vertebrae. Both injuries were treated conservatively with Taylor braces. CONCLUSION: We highlight one of the potential hazards of this recreational activity, and the almost identical fracture pattern in this couple.

  18. Biophysical and RNA Interference Inhibitory Properties of Oligonucleotides Carrying Tetrathiafulvalene Groups at Terminal Positions

    Directory of Open Access Journals (Sweden)

    Sónia Pérez-Rentero


    Full Text Available Oligonucleotide conjugates carrying a single functionalized tetrathiafulvalene (TTF unit linked through a threoninol molecule to the 3′ or 5′ ends were synthesized together with their complementary oligonucleotides carrying a TTF, pyrene, or pentafluorophenyl group. TTF-oligonucleotide conjugates formed duplexes with higher thermal stability than the corresponding unmodified oligonucleotides and pyrene- and pentafluorophenyl-modified oligonucleotides. TTF-modified oligonucleotides are able to bind to citrate-stabilized gold nanoparticles (AuNPs and produce stable gold AuNPs functionalized with oligonucleotides. Finally, TTF-oligoribonucleotides have been synthesized to produce siRNA duplexes carrying TTF units. The presence of the TTF molecule is compatible with the RNA interference mechanism for gene inhibition.

  19. 75 FR 61508 - Agency Information Collection Activities: Canadian Border Boat Landing Permit (CBP Form I-68) (United States)


    ... Landing Permit (CBP Form I-68) AGENCY: U.S. Customs and Border Protection (CBP), Department of Homeland... concerning the Canadian Border Boat Landing Permit (Form I- ] 68). This request for comment is being made... Form I-68. Abstract: The Canadian Border Boat Landing Permit (CBP Form I-68) allows...

  20. 78 FR 73875 - Agency Information Collection Activities: Canadian Border Boat Landing Permit (CBP Form I-68) (United States)


    ... Landing Permit (CBP Form I-68) AGENCY: U.S. Customs and Border Protection (CBP), Department of Homeland... requirement concerning the Canadian Border Boat Landing Permit (Form I-68). This request for comment is being... Form I-68. Abstract: The Canadian Border Boat Landing Permit (CBP Form I-68) allows...

  1. 76 FR 7701 - Special Local Regulations; Krewe of Charleston Mardi Gras Boat Parade, Charleston Harbor... (United States)


    ... SECURITY Coast Guard 33 CFR Part 100 RIN 1625-AA08 Special Local Regulations; Krewe of Charleston Mardi... Mardi Gras Boat Parade on the Ashley River and in Charleston Harbor in Charleston, South Carolina. This... Guard did not receive notice of the Krewe of Charleston Mardi Gras Boat Parade with sufficient time...

  2. 46 CFR 116.1120 - Drainage of cockpit vessels, well deck vessels, and open boats. (United States)


    ... 46 Shipping 4 2010-10-01 2010-10-01 false Drainage of cockpit vessels, well deck vessels, and open boats. 116.1120 Section 116.1120 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL... Drainage of cockpit vessels, well deck vessels, and open boats. Drainage of cockpit vessels, well...

  3. 33 CFR 100.109 - Winter Harbor Lobster Boat Race, Winter Harbor, ME. (United States)


    ... 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... Lobster Boat Race, Winter Harbor, ME. (a) Regulated area. The regulated area includes all waters of...

  4. 33 CFR 100.118 - Searsport Lobster Boat Races, Searsport Harbor, ME. (United States)


    ... 33 Navigation and Navigable Waters 1 2010-07-01 2010-07-01 false Searsport Lobster Boat Races, Searsport Harbor, ME. 100.118 Section 100.118 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF... Lobster Boat Races, Searsport Harbor, ME. (a) Regulated Area. The regulated area includes all waters...

  5. 33 CFR 100.909 - Chinatown Chamber of Commerce Dragon Boat Race; Chicago, IL. (United States)


    ... 33 Navigation and Navigable Waters 1 2010-07-01 2010-07-01 false Chinatown Chamber of Commerce Dragon Boat Race; Chicago, IL. 100.909 Section 100.909 Navigation and Navigable Waters COAST GUARD... Chinatown Chamber of Commerce Dragon Boat Race; Chicago, IL. (a) Regulated Area. All waters of the...

  6. 33 CFR 100.1302 - Special Local Regulation, Annual Dragon Boat Races, Portland, Oregon. (United States)


    ... 33 Navigation and Navigable Waters 1 2010-07-01 2010-07-01 false Special Local Regulation, Annual Dragon Boat Races, Portland, Oregon. 100.1302 Section 100.1302 Navigation and Navigable Waters COAST... § 100.1302 Special Local Regulation, Annual Dragon Boat Races, Portland, Oregon. (a) Regulated area....

  7. 33 CFR 100.903 - Harborfest Dragon Boat Race; South Haven, MI. (United States)


    ... 33 Navigation and Navigable Waters 1 2010-07-01 2010-07-01 false Harborfest Dragon Boat Race; South Haven, MI. 100.903 Section 100.903 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF... Dragon Boat Race; South Haven, MI. (a) Regulated Area. A regulated area is established to include...

  8. A traditional boats and navigational history of Odisha, East coast of India

    Digital Repository Service at National Institute of Oceanography (India)

    Tripati, S.

    -328. Greenhill, B. 1971. Boats and Boatmen of Pakistan, Newton Abbott: David and Charles. 14    Guy, J. 1999. A Boat Model and State Ritual in Eastern India, Bulletin de l'Ecole française d'Extrême-Orient 86: 105-126. Hill, A. H. 1958. Some early accounts...

  9. 76 FR 78151 - Special Local Regulations; Boca Raton Holiday Boat Parade, Intracoastal Waterway, Boca Raton, FL (United States)


    ... SECURITY Coast Guard 33 CFR Part 100 RIN 1625-AA08 Special Local Regulations; Boca Raton Holiday Boat Parade, Intracoastal Waterway, Boca Raton, FL AGENCY: Coast Guard, DHS. ACTION: Temporary final rule... Waterway in Boca Raton, Florida during the Boca Raton Holiday Boat Parade on Saturday, December 17,...

  10. First steps towards conformationally selective artificial lectins: the chair-boat discrimination by molecularly imprinted polymers. (United States)

    de Talancé, Vincent Lemau; Massinon, Olivier; Baati, Rachid; Wagner, Alain; Vincent, Stéphane P


    A series of molecularly imprinted polymers (MIPs) were prepared in the presence of a synthetic galactoside locked in a (1,4)B boat conformation. This study demonstrates that, depending on the polymerisation technique, an organic material can selectively bind a carbohydrate in a biologically relevant boat conformation.

  11. Has aid for AIDS raised all health funding boats? (United States)

    Shiffman, Jeremy; Berlan, David; Hafner, Tamara


    Global health analysts have debated whether donor prioritization of HIV/AIDS control has lifted all boats, raising attention and funding levels for health issues aside from HIV/AIDS. We investigate this question, considering donor funding for 4 historically prominent health agendas-HIV/AIDS, health systems strengthening, population and reproductive health, and infectious disease control-over the decade 1998-2007. We employ funding data from the Development Assistance Committee of the Organization for Economic Cooperation and Development, which tracks donor aid. The data indicate that HIV/AIDS may have helped to increase funding for the control of other infectious diseases; however, there is no firm evidence that other health issues beyond the control of infectious diseases have benefited. Between 1998 and 2007, funding for HIV/AIDS control rose from just 5.5% to nearly half of all aid for health. Over the same period, funding for health systems strengthening declined from 62.3% to 23.9% of total health aid and that for population and reproductive health declined from 26.4% to 12.3%. Also, even as total aid for health tripled during this decade, aid for health systems strengthening largely stagnated. Overall, the data indicate little support for the contention that donor funding for HIV/AIDS has lifted all boats.

  12. Inflation Basics

    Energy Technology Data Exchange (ETDEWEB)

    Green, Dan [Fermi National Accelerator Lab. (FNAL), Batavia, IL (United States)


    inflation since metrical fluctuations, both scalar and tensor, are also produced in inflationary models. Thus, the time appears to be appropriate for a very basic and simple exposition of the inflationary model written from a particle physics perspective. Only the simplest scalar model will be explored because it is easy to understand and contains all the basic elements of the inflationary model.

  13. Inflation Basics

    Energy Technology Data Exchange (ETDEWEB)

    Green, Dan [Fermi National Accelerator Lab. (FNAL), Batavia, IL (United States)


    inflation since metrical fluctuations, both scalar and tensor, are also produced in inflationary models. Thus, the time appears to be appropriate for a very basic and simple exposition of the inflationary model written from a particle physics perspective. Only the simplest scalar model will be explored because it is easy to understand and contains all the basic elements of the inflationary model.

  14. Quagga and zebra mussel risk via veliger transfer by overland hauled boats

    Directory of Open Access Journals (Sweden)

    Larry B. Dalton


    Full Text Available Invasive quagga and zebra mussels (Dreissena rostriformis bugensis and Dreissena polymorpha, respectively pose a great threat to USwaters. Recreational boats constitute a significant risk for spreading the organisms. Recreational boats circulate large amounts of raw waterwhen in use, and if not drained and dried correctly can transport many mussel larvae, called veligers. Veligers experience very high mortality rates; however, the number of potentially transported veligers can be a serious risk to non-infested bodies of water, especially if multiple boats are involved. The risk of veliger transport was calculated for Lake Mead and Lake Michigan using boat capacities for water circulation and specific veliger density data. Results illustrate the importance of draining, drying, and/or decontaminating recreational boats after use.

  15. Design and realization a skiff racing boat hull made of natural fibers reinforced composite (United States)

    Collotta, M.; Solazzi, L.; Pandini, S.; Tomasoni, G.; Alberti, M.; Donzella, G.


    This paper discusses the development of a racing boat with an hull made of a composite material reinforced by natural fibers. In particular, we report here the design and realization of the boat hull, the assessment of its mechanical performance by means of a computer assisted simulation, and the cost analysis to assess the economic sustainability of the new composite developed. The results have shown that the new composite has a performance comparable with conventional glass fiber reinforced composites employed for the realization of this type of boat, accordingly to the technology employed and the lamination sequence adopted. Moreover, the FEM analysis performed over the skiff of the designed and constructed boat has demonstrated a successful choice of the material for real application, as it was later confirmed by the good performance of the boat in water. Finally, the cost analysis highlighted the economic sustainability of the new composite, allowing a cost saving of over 28% with respect to carbon fiber composites.

  16. Typing of enteroviruses by use of microwell oligonucleotide arrays. (United States)

    Susi, P; Hattara, L; Waris, M; Luoma-Aho, T; Siitari, H; Hyypiä, T; Saviranta, P


    We have developed a straightforward assay for the rapid typing of enteroviruses using oligonucleotide arrays in microtiter wells. The viral nucleic acids are concomitantly amplified and labeled during reverse transcription-PCR, and unpurified PCR products are used for hybridization. DNA strands are separated by alkaline denaturation, and hybridization is started by neutralization. The microarray hybridization reactions and the subsequent washes are performed in standard 96-well microtiter plates, which makes the method easily adaptable to high-throughput analysis. We describe here the assay principle and its potential in clinical laboratory use by correctly identifying 10 different enterovirus reference strains. Furthermore, we explore the detection of unknown sequence variants using serotype consensus oligonucleotide probes. With just two consensus probes for the coxsackievirus A9 (CVA9) serotype, we detected 23 out of 25 highly diverse CVA9 isolates. Overall, the assay involves several features aiming at ease of performance, robustness, and applicability to large-scale studies.

  17. Solid-phase synthesis of siRNA oligonucleotides. (United States)

    Beaucage, Serge L


    Since the discovery of RNA interference (RNAi) as a means to silence the expression of specific genes, small interfering RNA (siRNA) oligonucleotides have been recognized as powerful tools for targeting therapeutically important mRNAs and eliciting their destruction. This discovery has created a high demand for synthetic oligoribonucleotides as potential therapeutics and has spurred a renaissance in the development of rapid, efficient methods for solid-phase RNA synthesis. The design and implementation of 2'-hydroxyl protecting groups that provide ribonucleoside phosphoramidites with coupling kinetics and coupling efficiencies comparable to those of deoxyribonucleoside phosphoramidites are key to the production of RNA oligonucleotides in sufficient quantity and purity for pharmaceutical applications. In this context, various siRNAs were chemically modified to identify the biophysical and biochemical parameters necessary for effective and stable RNAi-mediated gene-silencing activities.

  18. Electrochemical uranyl cation biosensor with DNA oligonucleotides as receptor layer. (United States)

    Jarczewska, Marta; Ziółkowski, Robert; Górski, Łukasz; Malinowska, Elżbieta


    The present study aims at the further development of the uranyl oligonucleotide-based voltammetric biosensor, which takes advantage of strong interaction between UO2(2+) and phosphate DNA backbone. Herein we report the optimization of working parameters of previously elaborated electrochemical DNA biosensor. It is shown that the sensor sensitivity is highly dependent on the oligonucleotide probe length and the incubation time of sensor in a sample solution. Consequently, the highest sensitivity was obtained for 10-nucleotide sequence and 60 min incubation time. The lower detection limit towards uranyl cation for developed biosensor was 30 nM. The influence of mixed monolayers and the possibility of developing a non-calibration device were also investigated. The selectivity of the proposed biosensor was significantly improved via elimination of adenine nucleobases from the DNA probe. Moreover, the regeneration procedure was elaborated and tested to prolong the use of the same biosensor for 4 subsequent determinations of UO2(2+).

  19. Palladium-Catalyzed Modification of Unprotected Nucleosides, Nucleotides, and Oligonucleotides

    Directory of Open Access Journals (Sweden)

    Kevin H. Shaughnessy


    Full Text Available Synthetic modification of nucleoside structures provides access to molecules of interest as pharmaceuticals, biochemical probes, and models to study diseases. Covalent modification of the purine and pyrimidine bases is an important strategy for the synthesis of these adducts. Palladium-catalyzed cross-coupling is a powerful method to attach groups to the base heterocycles through the formation of new carbon-carbon and carbon-heteroatom bonds. In this review, approaches to palladium-catalyzed modification of unprotected nucleosides, nucleotides, and oligonucleotides are reviewed. Polar reaction media, such as water or polar aprotic solvents, allow reactions to be performed directly on the hydrophilic nucleosides and nucleotides without the need to use protecting groups. Homogeneous aqueous-phase coupling reactions catalyzed by palladium complexes of water-soluble ligands provide a general approach to the synthesis of modified nucleosides, nucleotides, and oligonucleotides.

  20. One-oligonucleotide method for constructing vectors for RNA interference

    Institute of Scientific and Technical Information of China (English)



    Aim: To develop an easy, fast, automated, and inexpensive method for constructing short-hairpin-RNA cassettes for RNAi studies. Methods: Using single oligonucleotides, a variety of DNA cassettes for RNAi vectors were constructed in only few minutes in an automated manner. The cassettes, targeting the eGFP,were cloned into plasmids driven by RNA polymerase Ⅲ promoter H 1. Then, the plasmids were transfected into HeLa cells that were later infected with a recombinant adenovirus encoding the eGFP gene. The level of eGFP fluorescence was evaluated by confocal imaging and flow cytometry. Results: The plasmids constructed with the DNA cassettes made by the one-oligonucleotide method inhibited eGFP with different potencies, ranging from 55% to 75%. Conclusion: By using the method reported here, it is possible to simultaneously construct hundreds of different DNA cassettes for RNAi experiments in an inexpensive, automated way. This method will facilitate functional genomics studies on mammalian cells.

  1. Fluorescence quenching of TMR by guanosine in oligonucleotides

    Institute of Scientific and Technical Information of China (English)


    Nucleotide-specific fluorescence quenching in fluorescently labeled DNA has many applications in biotechnology. We have studied the inter-and intra-molecular quenching of tetramethylrhodamine (TMR) by nucleotides to better understand their quenching mechanism and influencing factors. In agreement with previous work, dGMP can effectively quench TMR, while the quenching of TMR by other nucleotides is negligible. The Stern-Volmer plot between TMR and dGMP delivers a bimolecular quenching constant of Ks=52.3 M-1. The fluorescence of TMR in labeled oligonucleotides decreases efficiently through photoinduced electron transfer by guanosine. The quenching rate constant between TMR and guanosine was measured using fluorescence correlation spectroscopy (FCS). In addition, our data show that the steric hindrance by bases around guanosine has significant effect on the G-quenching. The availability of these data should be useful in designing fluorescent oligonucleotides and understanding the G-quenching process.

  2. Voltammetric behaviour of oligonucleotide lipoplexes adsorbed onto glassy carbon electrodes


    Piedade, J. A. P.; M. Mano; Lima, M. C. Pedroso de; Oretskaya, T S; Oliveira-Brett, A. M.


    The voltammetric behaviour of oligonucleotide lipoplexes (ODN-lipoplexes) prepared from short oligodeoxynucleotides (ODN), with different base compositions, and liposomes of the cationic lipid DOTAP, was studied by differential pulse voltammetry with a glassy carbon mini-electrode. It was found that the ODN base composition influences the ODN-lipoplex voltammetric response. Differential pulse voltammograms for ODN-lipoplexes of the ODN adenosine nucleotides present two different features when...

  3. Anti-tumor activity of splice-switching oligonucleotides


    Bauman, John A; Li, Shyh-Dar; Yang, Angela; Huang, Leaf; Kole, Ryszard


    Alternative splicing has emerged as an important target for molecular therapies. Splice-switching oligonucleotides (SSOs) modulate alternative splicing by hybridizing to pre-mRNA sequences involved in splicing and blocking access to the transcript by splicing factors. Recently, the efficacy of SSOs has been established in various animal disease models; however, the application of SSOs against cancer targets has been hindered by poor in vivo delivery of antisense therapeutics to tumor cells. T...

  4. Thermodynamic treatment of oligonucleotide duplex–simplex equilibria (United States)

    Owczarzy, Richard; Dunietz, Isard; Behlke, Mark A.; Klotz, Irving M.; Walder, Joseph A.


    Thermodynamic formulations have been devised to obtain ΔG° values directly from spectroscopic data at a fixed common temperature in nucleic acid duplex–simplex melting curves. In addition, the dependence of melting on salt concentration has been expressed in terms of a stepwise stoichiometric representation, which leads to a specific equation for the partition of the added sodium ions between the different oligonucleotide forms. PMID:14657395

  5. Sex determination of bovine preimplantation embryos by oligonucleotide microarray. (United States)

    Yang, Hua; Zhong, Fagang; Yang, Yonglin; Wang, Xinhua; Liu, Shouren; Zhu, Bin


    The aim has been to set up a rapid and accurate microarray assay using sandwich mode for sex determination of bovine preimplantation embryos. Twelve sequence-specific oligonucleotide capture probes used to discriminate 12 samples were spotted onto the aldehyde-modified glass slides by Arrayer. The 2 recognition probes used to identify coding regions of the sex-determining region of the Y chromosome gene (SRY) and β-casein (CSN2) reference gene were coupled with biotin. The assay was optimized by using genomic DNA extracted from blood samples of known sex individuals. Polymerase chain reaction (PCR) was used to amplify the fragments in the HMG box region of SRY gene and CSN2 gene with sequence-specific primers. The sex of samples was identified by detecting both the SRY and CSN2 genes simultaneously in 2 reaction cells of microarrays, with the male having SRY and CSN2 signals and the female only CSN2. The sex of 20 bovine preimplantation embryos was determined by oligonucleotide microarray. The protocol was run with a blind test that showed a 100% (82/82) specificity and accuracy in sexing of leukocytes. The bovine embryos were transferred into 20 bovine recipients, with a pregnant rate of 40% (8/20). Three calves were born at term, and 5 fetuses were miscarried. Their sexes were fully in accordance with the embryonic sex predetermination predicted by oligonucleotide microarray. This suggests that the oligonucleotide microarray method of SRY gene analysis can be used in early sex prediction of bovine embryos in breeding programs.

  6. Thermoplastic polymers surfaces for Dip-Pen Nanolithography of oligonucleotides

    Energy Technology Data Exchange (ETDEWEB)

    Suriano, Raffaella [Dipartimento di Chimica, Materiali e Ingegneria Chimica “Giulio Natta”, Politecnico di Milano, Piazza Leonardo da Vinci 32, 20133 Milano (Italy); Biella, Serena, E-mail: [Dipartimento di Chimica, Materiali e Ingegneria Chimica “Giulio Natta”, Politecnico di Milano, Piazza Leonardo da Vinci 32, 20133 Milano (Italy); Cesura, Federico; Levi, Marinella; Turri, Stefano [Dipartimento di Chimica, Materiali e Ingegneria Chimica “Giulio Natta”, Politecnico di Milano, Piazza Leonardo da Vinci 32, 20133 Milano (Italy)


    Different thermoplastic polymers were spin-coated to prepare smooth surfaces for the direct deposition of end-group modified oligonucleotides by Dip-Pen Nanolithography. A study of the diffusion process was done in order to investigate the dependence of calibration coefficient and quality of deposited features on environmental parameters (temperature, relative humidity) and ink's molecular weight and functionality. The optimization of the process parameters led to the realization of high quality and density nanoarrays on plastics.

  7. ERT basics

    Energy Technology Data Exchange (ETDEWEB)

    Butters, M. [MBC Energy and Environment, Ottawa, ON (Canada)]|[National Round Table on the Environment and the Economy, Ottawa, ON (Canada)


    ERT is an economic instrument which helps power companies achieve emission reduction compliance cost-effectively. This paper presents the basics of ERT with reference to trading concepts, types of systems and types of emissions. The paper also describes the state of the Canadian energy market regarding greenhouse gases (GHG), nitrogen oxides, sulphur dioxide and volatile organic compounds. The association between ERT and district energy is also explained. By 2010, the global market for GHG trading is expected to be worth $10 billion to $3 trillion U.S. Canada has committed to reducing its GHG to 6 per cent below 1990 levels by 2012, but currently emits 705 Mt per year. This is expected to increase to 770 Mt by 2010. Therefore, in order to meet its commitment, GHGs will have to be reduced 200 Mt per year. Canada is currently considering ratifying the Kyoto agreement and a trading system is being developed. There are several abatement technologies currently under consideration for district energy systems, including adding scrubbers, improving efficiency, and fuel switching. The marginal cost of abatement was also discussed. tabs., figs.

  8. 78 FR 25574 - Special Local Regulations; Third Annual Space Coast Super Boat Grand Prix, Atlantic Ocean; Cocoa... (United States)


    ... Super Boat Grand Prix, Atlantic Ocean; Cocoa Beach, FL AGENCY: Coast Guard, DHS. ACTION: Temporary final... Ocean east of Cocoa Beach, Florida during the Space Coast Super Boat Grand Prix, a series of high-speed... Ocean east of Cocoa Beach, Florida. Approximately 30 high-speed power boats are anticipated...

  9. Particle-Based Microarrays of Oligonucleotides and Oligopeptides. (United States)

    Nesterov-Mueller, Alexander; Maerkle, Frieder; Hahn, Lothar; Foertsch, Tobias; Schillo, Sebastian; Bykovskaya, Valentina; Sedlmayr, Martyna; Weber, Laura K; Ridder, Barbara; Soehindrijo, Miriam; Muenster, Bastian; Striffler, Jakob; Bischoff, F Ralf; Breitling, Frank; Loeffler, Felix F


    In this review, we describe different methods of microarray fabrication based on the use of micro-particles/-beads and point out future tendencies in the development of particle-based arrays. First, we consider oligonucleotide bead arrays, where each bead is a carrier of one specific sequence of oligonucleotides. This bead-based array approach, appearing in the late 1990s, enabled high-throughput oligonucleotide analysis and had a large impact on genome research. Furthermore, we consider particle-based peptide array fabrication using combinatorial chemistry. In this approach, particles can directly participate in both the synthesis and the transfer of synthesized combinatorial molecules to a substrate. Subsequently, we describe in more detail the synthesis of peptide arrays with amino acid polymer particles, which imbed the amino acids inside their polymer matrix. By heating these particles, the polymer matrix is transformed into a highly viscous gel, and thereby, imbedded monomers are allowed to participate in the coupling reaction. Finally, we focus on combinatorial laser fusing of particles for the synthesis of high-density peptide arrays. This method combines the advantages of particles and combinatorial lithographic approaches.

  10. Particle-Based Microarrays of Oligonucleotides and Oligopeptides

    Directory of Open Access Journals (Sweden)

    Alexander Nesterov-Mueller


    Full Text Available In this review, we describe different methods of microarray fabrication based on the use of micro-particles/-beads and point out future tendencies in the development of particle-based arrays. First, we consider oligonucleotide bead arrays, where each bead is a carrier of one specific sequence of oligonucleotides. This bead-based array approach, appearing in the late 1990s, enabled high-throughput oligonucleotide analysis and had a large impact on genome research. Furthermore, we consider particle-based peptide array fabrication using combinatorial chemistry. In this approach, particles can directly participate in both the synthesis and the transfer of synthesized combinatorial molecules to a substrate. Subsequently, we describe in more detail the synthesis of peptide arrays with amino acid polymer particles, which imbed the amino acids inside their polymer matrix. By heating these particles, the polymer matrix is transformed into a highly viscous gel, and thereby, imbedded monomers are allowed to participate in the coupling reaction. Finally, we focus on combinatorial laser fusing of particles for the synthesis of high-density peptide arrays. This method combines the advantages of particles and combinatorial lithographic approaches.

  11. Characterization of self-assembled DNA concatemers from synthetic oligonucleotides

    Directory of Open Access Journals (Sweden)

    Lu Sun


    Full Text Available Studies of DNA–ligand interaction on a single molecule level provide opportunities to understand individual behavior of molecules. Construction of DNA molecules with repetitive copies of the same segments of sequences linked in series could be helpful for enhancing the interaction possibility for sequence-specific binding ligand to DNA. Here we report on the use of synthetic oligonucleotides to self-assembly into duplex DNA concatemeric molecules. Two strands of synthetic oligonucleotides used here were designed with 50-mer in length and the sequences are semi-complimentary so to hybridize spontaneously into concatemers of double stranded DNA. In order to optimize the length of the concatemers the oligonucleotides were incubated at different oligomer concentrations, ionic strengths and temperatures for different durations. Increasing the salt concentration to 200 mM NaCl was found to be the major optimizing factor because at this enhanced ionic strength the concatemers formed most quickly and the other parameters had no detectable effect. The size and shape of formed DNA concatemers were studied by gel electrophoresis in agarose, polyacrylamide gels and by AFM. Our results show that linear DNA constructs up to several hundred base pairs were formed and could be separated from a substantial fraction of non-linear constructs.

  12. Targeting several CAG expansion diseases by a single antisense oligonucleotide.

    Directory of Open Access Journals (Sweden)

    Melvin M Evers

    Full Text Available To date there are 9 known diseases caused by an expanded polyglutamine repeat, with the most prevalent being Huntington's disease. Huntington's disease is a progressive autosomal dominant neurodegenerative disorder for which currently no therapy is available. It is caused by a CAG repeat expansion in the HTT gene, which results in an expansion of a glutamine stretch at the N-terminal end of the huntingtin protein. This polyglutamine expansion plays a central role in the disease and results in the accumulation of cytoplasmic and nuclear aggregates. Here, we make use of modified 2'-O-methyl phosphorothioate (CUGn triplet-repeat antisense oligonucleotides to effectively reduce mutant huntingtin transcript and protein levels in patient-derived Huntington's disease fibroblasts and lymphoblasts. The most effective antisense oligonucleotide, (CUG(7, also reduced mutant ataxin-1 and ataxin-3 mRNA levels in spinocerebellar ataxia 1 and 3, respectively, and atrophin-1 in dentatorubral-pallidoluysian atrophy patient derived fibroblasts. This antisense oligonucleotide is not only a promising therapeutic tool to reduce mutant huntingtin levels in Huntington's disease but our results in spinocerebellar ataxia and dentatorubral-pallidoluysian atrophy cells suggest that this could also be applicable to other polyglutamine expansion disorders as well.


    Directory of Open Access Journals (Sweden)

    Pedro Miguel Duarte Moreno


    Full Text Available Under clinical development since the early 90’s and with two successfully approved drugs (Fomivirsen and Mipomersen, oligonucleotide-based therapeutics have not yet delivered a clinical drug to the market in the cancer field. Whilst many pre-clinical data has been generated, a lack of understanding still exists on how to efficiently tackle all the different challenges presented for cancer targeting in a clinical setting. Namely, effective drug vectorization, careful choice of target gene or synergistic multi-gene targeting are surely decisive, while caution must be exerted to avoid potential toxic, often misleading off-target-effects. Here a brief overview will be given on the nucleic acid chemistry advances that established oligonucleotide technologies as a promising therapeutic alternative and ongoing cancer related clinical trials. Special attention will be given towards a perspective on the hurdles encountered specifically in the cancer field by this class of therapeutic oligonucleotides and a view on possible avenues for success is presented, with particular focus on the contribution from nanotechnology to the field.

  14. G-Quadruplex Forming Oligonucleotides as Anti-HIV Agents. (United States)

    Musumeci, Domenica; Riccardi, Claudia; Montesarchio, Daniela


    Though a variety of different non-canonical nucleic acids conformations have been recognized, G-quadruplex structures are probably the structural motifs most commonly found within known oligonucleotide-based aptamers. This could be ascribed to several factors, as their large conformational diversity, marked responsiveness of their folding/unfolding processes to external stimuli, high structural compactness and chemo-enzymatic and thermodynamic stability. A number of G-quadruplex-forming oligonucleotides having relevant in vitro anti-HIV activity have been discovered in the last two decades through either SELEX or rational design approaches. Improved aptamers have been obtained by chemical modifications of natural oligonucleotides, as terminal conjugations with large hydrophobic groups, replacement of phosphodiester linkages with phosphorothioate bonds or other surrogates, insertion of base-modified monomers, etc. In turn, detailed structural studies have elucidated the peculiar architectures adopted by many G-quadruplex-based aptamers and provided insight into their mechanism of action. An overview of the state-of-the-art knowledge of the relevance of putative G-quadruplex forming sequences within the viral genome and of the most studied G-quadruplex-forming aptamers, selectively targeting HIV proteins, is here presented.

  15. Recursive construction of perfect DNA molecules from imperfect oligonucleotides. (United States)

    Linshiz, Gregory; Yehezkel, Tuval Ben; Kaplan, Shai; Gronau, Ilan; Ravid, Sivan; Adar, Rivka; Shapiro, Ehud


    Making faultless complex objects from potentially faulty building blocks is a fundamental challenge in computer engineering, nanotechnology and synthetic biology. Here, we show for the first time how recursion can be used to address this challenge and demonstrate a recursive procedure that constructs error-free DNA molecules and their libraries from error-prone oligonucleotides. Divide and Conquer (D&C), the quintessential recursive problem-solving technique, is applied in silico to divide the target DNA sequence into overlapping oligonucleotides short enough to be synthesized directly, albeit with errors; error-prone oligonucleotides are recursively combined in vitro, forming error-prone DNA molecules; error-free fragments of these molecules are then identified, extracted and used as new, typically longer and more accurate, inputs to another iteration of the recursive construction procedure; the entire process repeats until an error-free target molecule is formed. Our recursive construction procedure surpasses existing methods for de novo DNA synthesis in speed, precision, amenability to automation, ease of combining synthetic and natural DNA fragments, and ability to construct designer DNA libraries. It thus provides a novel and robust foundation for the design and construction of synthetic biological molecules and organisms.

  16. Differentiation of regions with atypical oligonucleotide composition in bacterial genomes

    Directory of Open Access Journals (Sweden)

    Reva Oleg N


    Full Text Available Abstract Background Complete sequencing of bacterial genomes has become a common technique of present day microbiology. Thereafter, data mining in the complete sequence is an essential step. New in silico methods are needed that rapidly identify the major features of genome organization and facilitate the prediction of the functional class of ORFs. We tested the usefulness of local oligonucleotide usage (OU patterns to recognize and differentiate types of atypical oligonucleotide composition in DNA sequences of bacterial genomes. Results A total of 163 bacterial genomes of eubacteria and archaea published in the NCBI database were analyzed. Local OU patterns exhibit substantial intrachromosomal variation in bacteria. Loci with alternative OU patterns were parts of horizontally acquired gene islands or ancient regions such as genes for ribosomal proteins and RNAs. OU statistical parameters, such as local pattern deviation (D, pattern skew (PS and OU variance (OUV enabled the detection and visualization of gene islands of different functional classes. Conclusion A set of approaches has been designed for the statistical analysis of nucleotide sequences of bacterial genomes. These methods are useful for the visualization and differentiation of regions with atypical oligonucleotide composition prior to or accompanying gene annotation.

  17. Synthesis of triazole-linked oligonucleotides with high affinity to DNA complements and an analysis of their compatibility with biosystems. (United States)

    Varizhuk, Anna M; Kaluzhny, Dmitry N; Novikov, Roman A; Chizhov, Alexandr O; Smirnov, Igor P; Chuvilin, Andrey N; Tatarinova, Olga N; Fisunov, Gleb Y; Pozmogova, Galina E; Florentiev, Vladimir L


    New oligonucleotide analogues with triazole internucleotide linkages were synthesized, and their hybridization properties were studied. The analogues demonstrated DNA binding affinities similar to those of unmodified oligonucleotides. The modification was shown to protect the oligonucleotides from nuclease hydrolysis. The modified oligonucleotides were tested as PCR primers. Modifications remote from the 3'-terminus were tolerated by polymerases. Our results suggest that these new oligonucleotide analogues are among the most promising triazole DNA mimics characterized to date.

  18. A new oligonucleotide microarray for detection of pathogenic and non-pathogenic Legionella spp. (United States)

    Cao, Boyang; Liu, Xiangqian; Yu, Xiang; Chen, Min; Feng, Lu; Wang, Lei


    Legionella pneumophila has been recognized as the major cause of legionellosis since the discovery of the deadly disease. Legionella spp. other than L. pneumophila were later found to be responsible to many non-pneumophila infections. The non-L. pneumophila infections are likely under-detected because of a lack of effective diagnosis. In this report, we have sequenced the 16S-23S rRNA gene internal transcribed spacer (ITS) of 10 Legionella species and subspecies, including L. anisa, L. bozemanii, L. dumoffii, L. fairfieldensis, L. gormanii, L. jordanis, L. maceachernii, L. micdadei, L. pneumophila subspp. fraseri and L. pneumophila subspp. pasculleii, and developed a rapid oligonucleotide microarray detection technique accordingly to identify 12 most common Legionella spp., which consist of 11 pathogenic species of L. anisa, L. bozemanii, L. dumoffii, L. gormanii, L. jordanis, L. longbeachae, L. maceachernii, L. micdadei, and L. pneumophila (including subspp. pneumophila, subspp. fraseri, and subspp. pasculleii) and one non-pathogenic species, L. fairfieldensis. Twenty-nine probes that reproducibly detected multiple Legionella species with high specificity were included in the array. A total of 52 strains, including 30 target pathogens and 22 non-target bacteria, were used to verify the oligonucleotide microarray assay. The sensitivity of the detection was at 1.0 ng with genomic DNA or 13 CFU/100 mL with Legionella cultures. The microarray detected seven samples of air conditioner-condensed water with 100% accuracy, validating the technique as a promising method for applications in basic microbiology, clinical diagnosis, food safety, and epidemiological surveillance. The phylogenetic study based on the ITS has also revealed that the non-pathogenic L. fairfieldensis is the closest to L. pneumophila than the nine other pathogenic Legionella spp.

  19. Evidence for Aedes aegypti (Diptera: Culicidae) Oviposition on Boats in the Peruvian Amazon. (United States)

    Guagliardo, Sarah Anne; Morrison, Amy C; Luis Barboza, Jose; Wesson, Dawn M; Ponnusamy, Loganathan; Astete, Helvio; Vazquez-Prokopec, Gonzalo; Kitron, Uriel


    Dengue vector Aedes aegypti L. is invading peri-urban and rural areas throughout Latin America. Our previous research in the Peruvian Amazon has shown that river boats are heavily infested with immature and adult Ae. aegypti mosquitoes, likely playing a major role in their long-distance dispersal and successful invasion. However, the presence of immature mosquitoes provides no information about the timing of oviposition, and whether it took place in the boats. Here, we used baited ovitraps deployed on river boats to test the hypothesis that Ae. aegypti oviposition occurs during boat travel. We deployed 360 ovitraps on 60 different barges during August and October of 2013, and February 2014 (with 20 barges sampled during each month). We found that Ae. aegypti mosquitoes in 22 individual ovitraps from 15 of the 60 barges (premise index 25%) across all sampling dates. Further, the distribution of Ae. aegypti egg abundance was highly aggregated: 2.6% of traps (N=7) were responsible for 71.8% of eggs found, and 1.5% of traps (N=4) were responsible for all (100%) of the larvae found. Similarly, 5% of boats were responsible for the 71.47% of eggs. Our results provide strong evidence that Ae. aegypti oviposition commonly occurs during boat travel. Baited ovitraps could represent a cost-effective means of monitoring and controlling mosquito populations on boats.

  20. Effect of boat noise and angling on lake fish behaviour

    DEFF Research Database (Denmark)

    Jacobsen, Lene; Baktoft, Henrik; Jepsen, Niels;


    The effects of disturbances from recreational activities on the swimming speed and habitat use of roach Rutilus rutilus, perch Perca fluviatilis and pike Esox lucius were explored. Disturbances were applied for 4h as (1) boating in short intervals with a small outboard internal combustion engine...... was significantly higher during disturbances [both (1) and (2)] with an immediate reaction shortly after the engine started. Perca fluviatilis displayed increased swimming activity during the first hour of disturbance but not during the following hours. Swimming activity of E. lucius was not significantly different...... between disturbance periods and the same periods on days without disturbance (control). Rutilus rutilus increased their use of the central part of the lake during disturbances, whereas no habitat change was observed in P. fluviatilis and E. lucius. No difference in fish response was detected between...

  1. Experimental Evaluation of the Hydrodynamic Response of Crew Boat Hulls

    DEFF Research Database (Denmark)

    Pecher, Arthur; Kofoed, Jens Peter

    This report presents the results of an experimental study on 3 different hulls of crew boats intended for service to offshore wind turbines. Their hydrodynamic behaviour has been tested in various sea states representing general wave conditions that could be expected at offshore wind farms. Two...... main setups were used during the tests, corresponding to the hulls being connected to an offshore windmill and being free floating. The following aspects were the main subjects of investigation: • The Response Amplitude Operators of the hulls in two different configurations and with waves coming from 3...... the supervision of Jens Peter Kofoed in the deep wave basin in the Hydraulics and Coastal Engineering laboratory at Aalborg University during spring 2012....

  2. Discrimination of oligonucleotides of different lengths with a wild-type aerolysin nanopore (United States)

    Cao, Chan; Ying, Yi-Lun; Hu, Zheng-Li; Liao, Dong-Fang; Tian, He; Long, Yi-Tao


    Protein nanopores offer an inexpensive, label-free method of analysing single oligonucleotides. The sensitivity of the approach is largely determined by the characteristics of the pore-forming protein employed, and typically relies on nanopores that have been chemically modified or incorporate molecular motors. Effective, high-resolution discrimination of oligonucleotides using wild-type biological nanopores remains difficult to achieve. Here, we show that a wild-type aerolysin nanopore can resolve individual short oligonucleotides that are 2 to 10 bases long. The sensing capabilities are attributed to the geometry of aerolysin and the electrostatic interactions between the nanopore and the oligonucleotides. We also show that the wild-type aerolysin nanopores can distinguish individual oligonucleotides from mixtures and can monitor the stepwise cleavage of oligonucleotides by exonuclease I.

  3. Discussions on the possible origin of Europe's first boats - 11,500 BP

    Directory of Open Access Journals (Sweden)

    Peter Fletcher


    Full Text Available Three decades ago Detlev Ellmers suggested that hide (skin boats were being used to hunt swimming reindeer in glacial lakes of northern Germany's Upper Palaeolithic. This paper presents new material in support of Ellmers' contention. A paleo-osteological study from Stellmoor suggests that arrows were directed at the neck and upper shoulders from above and behind, at close range which can be considered evidence of them being shot by hunters in boats as the animals were swimming. Lyngby axes are linked to this hunting technique which provides possible distribution and dating for this proposed early boat technology.

  4. Synthesis of triazole-nucleoside phosphoramidites and their use in solid-phase oligonucleotide synthesis. (United States)

    Peel, Brandon J; Efthymiou, Tim C; Desaulniers, Jean-Paul


    Triazole-backbone oligonucleotides are macromolecules that have one or more triazole units that are acting as a backbone mimic. Triazoles within the backbone have been used within oligonucleotides for a variety of applications. This unit describes the preparation and synthesis of two triazole-nucleoside phosphoramidites [uracil-triazole-uracil (UtU) and cytosine-triazole-uracil (CtU)] based on a PNA-like scaffold, and their incorporation within oligonucleotides.

  5. Hemopoiesis-stimulating activity of immobilized oligonucleotides and hyaluronidase during cytostatic-induced myelosuppression. (United States)

    Dygai, A M; Skurikhin, E G; Pershina, O V; Zhdanov, V V; Khmelevskaya, A M; Andreeva, T V; Poponina, A M; Zjuzkov, G N; Udut, E V; Khrichkova, T Ju; Simanina, E V; Miroshnichenko, L A; Stavrova, L A; Tchaikovsky, A S; Markova, T S; Gurto, R V; Brjushinina, O S; Slepichev, V A


    The hemopoiesis-stimulating effect of combined treatment with immobilized oligonucleotides and hyaluronidase preparations was studied during cytostatic-induced myelosuppression caused by cyclophosphamide administration. Immobilized hyaluronidase was shown to increase the efficiency of correction of changes in the erythroid and granulocytic hemopoietic stems with immobilized oligonucleotides. This potentiation of the effect of immobilized oligonucleotides by immobilized hyaluronidase was related to an increase in functional activity of committed hemopoietic precursors.

  6. A New Achiral Linker Reagent for the Incorporation of Multiple Amino Groups Into Oligonucleotides

    DEFF Research Database (Denmark)


    The present invention relates to a new functionalized achiral linker reagent for incorporating multiple primary amino groups or reporter groups into oligonucleotides following the phosphoramidite methodology. It is possible to substitute any ribodeoxynucleotide, deoxynucleotide, or nucleotide wit......, to a method for preparing a labelled oligonucleotide, and to the use of the labelled oligonucleotide as hybridisation probe, in polymerase chain reactions (PCR), in nucleic acid sequencing, in cloning recombinant DNA and $i(in vitro) mutagenesis....

  7. Label-free detection of hybridization of oligonucleotides by oblique-incidence reflectivity difference method

    Institute of Scientific and Technical Information of China (English)


    The microarrays of 20-base oligonucleotide with different concentrations are detected before and after hybridization by the oblique-incidence reflectivity difference (OI-RD) method. The experimental results prove that OI-RD is a label-free method which can not only distinguish the concentration difference of oligonucleotides before and after the hybridization but also detect the hybridization of short oligonucleotides. At present the OI-RD method can detect 0.39 μmol/L 20-base oligonucleotide or less. These results suggest that the OI-RD method is a promising and potential technique for label-free detection of biological microarrays.

  8. Efficient assembly of very short oligonucleotides using T4 DNA Ligase

    Directory of Open Access Journals (Sweden)

    Holt Robert A


    Full Text Available Abstract Background In principle, a pre-constructed library of all possible short oligonucleotides could be used to construct many distinct gene sequences. In order to assess the feasibility of such an approach, we characterized T4 DNA Ligase activity on short oligonucleotide substrates and defined conditions suitable for assembly of a plurality of oligonucleotides. Findings Ligation by T4 DNA Ligase was found to be dependent on the formation of a double stranded DNA duplex of at least five base pairs surrounding the site of ligation. However, ligations could be performed effectively with overhangs smaller than five base pairs and oligonucleotides as small as octamers, in the presence of a second, complementary oligonucleotide. We demonstrate the feasibility of simultaneous oligonucleotide phosphorylation and ligation and, as a proof of principle for DNA synthesis through the assembly of short oligonucleotides, we performed a hierarchical ligation procedure whereby octamers were combined to construct a target 128-bp segment of the beta-actin gene. Conclusions Oligonucleotides as short as 8 nucleotides can be efficiently assembled using T4 DNA Ligase. Thus, the construction of synthetic genes, without the need for custom oligonucleotide synthesis, appears feasible.

  9. Splice-switching antisense oligonucleotides as therapeutic drugs


    Havens, Mallory A.; Hastings, Michelle L.


    Splice-switching oligonucleotides (SSOs) are short, synthetic, antisense, modified nucleic acids that base-pair with a pre-mRNA and disrupt the normal splicing repertoire of the transcript by blocking the RNA–RNA base-pairing or protein–RNA binding interactions that occur between components of the splicing machinery and the pre-mRNA. Splicing of pre-mRNA is required for the proper expression of the vast majority of protein-coding genes, and thus, targeting the process offers a means to manipu...

  10. Chemically modified oligonucleotides with efficient RNase H response

    DEFF Research Database (Denmark)

    Vester, Birte; Boel, Anne Marie; Lobedanz, Sune;


    Ten different chemically modified nucleosides were incorporated into short DNA strands (chimeric oligonucleotides ON3-ON12 and ON15-ON24) and then tested for their capacity to mediate RNAse H cleavage of the complementary RNA strand. The modifications were placed at two central positions directly...... in the RNase H cleaving region. The RNA strand of duplexes with ON3, ON5 and ON12 were cleaved more efficiently than the RNA strand of the DNA:RNA control duplex. There seems to be no correlation between the thermal stability between the duplexes and RNase H cleavage....

  11. A quantitative framework for estimating risk of collision between marine mammals and boats (United States)

    Martin, Julien; Sabatier, Quentin; Gowan, Timothy A.; Giraud, Christophe; Gurarie, Eliezer; Calleson, Scott; Ortega-Ortiz, Joel G.; Deutsch, Charles J.; Rycyk, Athena; Koslovsky, Stacie M.


    Speed regulations of watercraft in protected areas are designed to reduce lethal collisions with wildlife but can have economic consequences. We present a quantitative framework for investigating the risk of deadly collisions between boats and wildlife.

  12. Life-threatening neurovascular injuries associated with recreational use of "banana" boats. (United States)

    Cohen, José E; Moscovici, Samuel; Rosenthal, Guy; Benifla, Mony; Itshayek, Eyal


    Banana boat rides are a popular form of recreation worldwide. Recommendations that speed should not exceed 15 mph, passengers should wear protective gear, and an observer should be present on the towing boat are generally ignored. Medical personnel at tourist venues and general practitioners may not be attuned to the risk of serious injury. We present our experience in the management carotid- and vertebral artery dissections sustained by 44- and 23-year-old males during banana boat rides. In both cases, the dissections were misdiagnosed until patients presented to the Emergency Department two days after injury. In the first patient, medical management failed and endovascular treatment was required due to neurological deterioration. In patient two, anticoagulation therapy prevented embolic sequelae. Boat operators, passengers, and the medical personnel who are first to see these patients should be aware of the risk of arterial dissection to facilitate early detection of these potentially life-threatening injuries.

  13. Development of Algorithms for Control of Motor Boat as Multidimensional Nonlinear Object

    Directory of Open Access Journals (Sweden)

    Gaiduk Anatoliy


    Full Text Available In this paper authors develop and research system for motor boat control, that allows to move along the stated paths with the given speed. It is assumed, that boat is equipped by the measuring system that provides current coordinates, linear and angular velocities. Control system is based upon the mathematical model, presented earlier (see references. In order to analytically find the necessary controls, all equations were transformed to Jordan controllable form. Besides solution this transformation also allows to handle model nonlinearities and get required quality of movement along the stated paths. Control system includes algorithms for control of longtitudal velocity and boat course. Research of the proposed control system according to boat design limitations for the values of control variables was performed by simulation in MATLAB. Results of two experiments, different in value of the required velocity are discussed.

  14. The use of oligonucleotide probes for meningococcal serotype characterization

    Directory of Open Access Journals (Sweden)

    SACCHI Claudio Tavares


    Full Text Available In the present study we examine the potential use of oligonucleotide probes to characterize Neisseria meningitidis serotypes without the use of monoclonal antibodies (MAbs. Antigenic diversity on PorB protein forms the bases of serotyping method. However, the current panel of MAbs underestimated, by at least 50% the PorB variability, presumably because reagents for several PorB variable regions (VRs are lacking, or because a number of VR variants are not recognized by serotype-defining MAbs12. We analyzed the use of oligonucleotide probes to characterize serotype 10 and serotype 19 of N. meningitidis. The porB gene sequence for the prototype strain of serotype 10 was determined, aligned with 7 other porB sequences from different serotypes, and analysis of individual VRs were performed. The results of DNA probes 21U (VR1-A and 615U (VR3-B used against 72 N. meningitidis strains confirm that VR1 type A and VR3 type B encode epitopes for serotype-defined MAbs 19 and 10, respectively. The use of probes for characterizing serotypes possible can type 100% of the PorB VR diversity. It is a simple and rapid method specially useful for analysis of large number of samples.

  15. Oligonucleotide-Based Therapy for FTD/ALS Caused by the C9orf72 Repeat Expansion: A Perspective

    Directory of Open Access Journals (Sweden)

    Stephanie A. Fernandes


    Full Text Available Amyotrophic lateral sclerosis (ALS is a progressive and lethal disease of motor neuron degeneration, leading to paralysis of voluntary muscles and death by respiratory failure within five years of onset. Frontotemporal dementia (FTD is characterised by degeneration of frontal and temporal lobes, leading to changes in personality, behaviour, and language, culminating in death within 5–10 years. Both of these diseases form a clinical, pathological, and genetic continuum of diseases, and this link has become clearer recently with the discovery of a hexanucleotide repeat expansion in the C9orf72 gene that causes the FTD/ALS spectrum, that is, c9FTD/ALS. Two basic mechanisms have been proposed as being potentially responsible for c9FTD/ALS: loss-of-function of the protein encoded by this gene (associated with aberrant DNA methylation and gain of function through the formation of RNA foci or protein aggregates. These diseases currently lack any cure or effective treatment. Antisense oligonucleotides (ASOs are modified nucleic acids that are able to silence targeted mRNAs or perform splice modulation, and the fact that they have proved efficient in repeat expansion diseases including myotonic dystrophy type 1 makes them ideal candidates for c9FTD/ALS therapy. Here, we discuss potential mechanisms and challenges for developing oligonucleotide-based therapy for c9FTD/ALS.

  16. Oligonucleotide-Based Therapy for FTD/ALS Caused by the C9orf72 Repeat Expansion: A Perspective. (United States)

    Fernandes, Stephanie A; Douglas, Andrew G L; Varela, Miguel A; Wood, Matthew J A; Aoki, Yoshitsugu


    Amyotrophic lateral sclerosis (ALS) is a progressive and lethal disease of motor neuron degeneration, leading to paralysis of voluntary muscles and death by respiratory failure within five years of onset. Frontotemporal dementia (FTD) is characterised by degeneration of frontal and temporal lobes, leading to changes in personality, behaviour, and language, culminating in death within 5-10 years. Both of these diseases form a clinical, pathological, and genetic continuum of diseases, and this link has become clearer recently with the discovery of a hexanucleotide repeat expansion in the C9orf72 gene that causes the FTD/ALS spectrum, that is, c9FTD/ALS. Two basic mechanisms have been proposed as being potentially responsible for c9FTD/ALS: loss-of-function of the protein encoded by this gene (associated with aberrant DNA methylation) and gain of function through the formation of RNA foci or protein aggregates. These diseases currently lack any cure or effective treatment. Antisense oligonucleotides (ASOs) are modified nucleic acids that are able to silence targeted mRNAs or perform splice modulation, and the fact that they have proved efficient in repeat expansion diseases including myotonic dystrophy type 1 makes them ideal candidates for c9FTD/ALS therapy. Here, we discuss potential mechanisms and challenges for developing oligonucleotide-based therapy for c9FTD/ALS.

  17. Studies on the Syntheses and Properties of 5'-Branched-sugar Isonucleosides and the Related Oligonucleotides

    Institute of Scientific and Technical Information of China (English)

    Tian Xiaobing; Zhang Lihe; Min Jimei


    @@ The chemistry of nucleosides and oligonucleotides is an actively investigated field in the search for new drugs. Thesyntheses and the properties of isonucleosides and oligonucleotides have been investigated to improve their stability,antitumor and antiviral activities, and to reduce their toxicity.

  18. The MOX/SUC precursor strategies: robust ways to construct functionalized oligonucleotides. (United States)

    Polushin, N


    The use of phosphoramidites bearing one or more methoxyoxalamido (MOX) or succinimido (SUC) reactive groups for construction of functionalized oligonucleotides is described. The efficiency of the new precursor strategy was demonstrated in the synthesis of oligonucleotide containing up to 16 imidazole residues.

  19. The big crossing: illegal boat migrants in the Mediterranean. (United States)

    Kassar, Hassène; Dourgnon, Paul


    This article explores illegal migration routes and groups across North Africa to Europe. We describe sub-Saharan and cross-Mediterranean routes, and how they changed during the years. We propose an analytical framework for the main factors for these migrations, from local to international and regulatory context. We then describe sea-migrants' nationalities and socio-economic and demographic characteristics, from studies undertook in Tunisia and Morocco. While boat migration represents only a fraction of illegal migration to Europe, it raises humanitarian as well as ethical issues for European and North African (NA) countries, as a non-negligible amount of them end up in death tolls of shipwrecks in the Mediterranean Sea. Moreover, existing statistics show that illegal trans-Mediterranean migration is growing exponentially. Ongoing crises in Africa and the Middle East are likely to prompt even larger outflows of refugees in the near future. This should induce NA countries to share closer public policy concerns with European countries.

  20. Detection of oligonucleotide hybridization on a single microparticle by time-resolved fluorometry: hybridization assays on polymer particles obtained by direct solid phase assembly of the oligonucleotide probes. (United States)

    Hakala, H; Heinonen, P; Iitiä, A; Lönnberg, H


    Oligodeoxyribonucleotides were assembled by conventional phosphoramidite chemistry on uniformly sized (50 microns) porous glycidyl methacrylate/ethylene dimethacrylate (SINTEF) and compact polystyrene (Dynosphere) particles, the aminoalkyl side chains of which were further derivatized with DMTrO-acetyl groups. The linker was completely resistant toward ammonolytic deprotection of the base moieties. The quality of oligonucleotides was assessed by repeating the synthesis on the same particles derivatized with a cleavable ester linker. The ability of the oligonucleotide-coated particles to bind complementary sequences via hybridization was examined by following the attachment of oligonucleotides bearing a photoluminescent europium(III) chelate to the particles. The fluorescence emission was measured directly on a single particle. The effects of the following factors on the kinetics and efficiency of hybridization were studied: number of particles in a given volume of the assay solution, loading of oligonucleotide on the particle, concentration of the target oligonucleotide in solution, length of the hybridizing sequence, presence of noncomplementary sequences, and ionic strength. The fluorescence signal measured on a single particle after hybridization was observed to be proportional to the concentration of the target oligonucleotide in solution over a concentration range of 5 orders of magnitude.

  1. Affinity hydrogels for controlled protein release using nucleic acid aptamers and complementary oligonucleotides. (United States)

    Soontornworajit, Boonchoy; Zhou, Jing; Snipes, Matthew P; Battig, Mark R; Wang, Yong


    Biomaterials for the precise control of protein release are important to the development of new strategies for treating human diseases. This study aimed to fundamentally understand aptamer--protein dissociation triggered by complementary oligonucleotides, and to apply this understanding to develop affinity hydrogels for controlled protein release. The results showed that the oligonucleotide tails of the aptamers played a critical role in inducing intermolecular hybridization and triggering aptamer--protein dissociation. In addition, the attachment of the oligonucleotide tails to the aptamers and the increase of hybridizing length could produce a synergistic effect on the dissociation of bound proteins from their aptamers. More importantly, pegylated complementary oligonucleotides could successfully trigger protein release from the aptamer-functionalized hydrogels at multiple time points. Based on these results, it is believed that aptamer-functionalized hydrogels and complementary oligonucleotides hold great potential of controlling the release of protein drugs to treat human diseases.

  2. Design of oligonucleotides for microarrays and perspectives for design of multi-transcriptome arrays

    DEFF Research Database (Denmark)

    Nielsen, Henrik Bjørn; Wernersson, Rasmus; Knudsen, Steen


    with an overview of these parameters. We present here a flexible tool named OligoWiz for designing oligonucleotides for multiple purposes. OligoWiz presents a set of parameter scores in a graphical interface to facilitate an overview for the user. Additional custom parameter scores can easily be added......Optimal design of oligonucleotides for microarrays involves tedious and laborious work evaluating potential oligonucleotides relative to a series of parameters. The currently available tools for this purpose are limited in their flexibility and do not present the oligonucleotide designer...... to the program to extend the default parameters: homology, DeltaTm, low-complexity, position and GATC-only. Furthermore we present an analysis of the limitations in designing oligonucleotide sets that can detect transcripts from multiple organisms. OligoWiz is available at

  3. Recommendations for safety pharmacology evaluations of oligonucleotide-based therapeutics. (United States)

    Berman, Cindy L; Cannon, Keri; Cui, Yi; Kornbrust, Douglas J; Lagrutta, Armando; Sun, Sunny Z; Tepper, Jeff; Waldron, Gareth; Younis, Husam S


    This document was prepared by the Safety Pharmacology Subcommittee of the Oligonucleotide Safety Working Group (OSWG), a group of industry and regulatory scientists involved in the development and regulation of therapeutic oligonucleotides. The mission of the Subcommittee was to develop scientific recommendations for the industry regarding the appropriate scope and strategies for safety pharmacology evaluations of oligonucleotides (ONs). These recommendations are the consensus opinion of the Subcommittee and do not necessarily reflect the current expectations of regulatory authorities. 1) Safety pharmacology testing, as described in the International Conference on Harmonisation (ICH) S7 guidance, is as applicable to ONs as it is to small molecule drugs and biotherapeutics. 2) Study design considerations for ONs are similar to those for other classes of drugs. In general, as with other therapeutics, studies should evaluate the drug product administered via the clinical route. Species selection should ideally consider relevance of the model with regard to the endpoints of interest, pharmacological responsiveness, and continuity with the nonclinical development program. 3) Evaluation of potential effects in the core battery (cardiovascular, central nervous, and respiratory systems) is recommended. In general: a. In vitro human ether-a-go-go-related gene (hERG) testing does not provide any specific value and is not warranted. b. Emphasis should be placed on in vivo evaluation of cardiovascular function, typically in nonhuman primates (NHPs). c. Due to the low level of concern, neurologic and respiratory function can be assessed concurrently with cardiovascular safety pharmacology evaluation in NHPs, within repeat-dose toxicity studies, or as stand-alone studies. In the latter case, rodents are most commonly used. 4) Other dedicated safety pharmacology studies, beyond the core battery, may have limited value for ONs. Although ONs can accumulate in the kidney and liver

  4. Enhanced gene repair mediated by methyl-CpG-modified single-stranded oligonucleotides (United States)

    Bertoni, Carmen; Rustagi, Arjun; Rando, Thomas A.


    Gene editing mediated by oligonucleotides has been shown to induce stable single base alterations in genomic DNA in both prokaryotic and eukaryotic organisms. However, the low frequencies of gene repair have limited its applicability for both basic manipulation of genomic sequences and for the development of therapeutic approaches for genetic disorders. Here, we show that single-stranded oligodeoxynucleotides (ssODNs) containing a methyl-CpG modification and capable of binding to the methyl-CpG binding domain protein 4 (MBD4) are able to induce >10-fold higher levels of gene correction than ssODNs lacking the specific modification. Correction was stably inherited through cell division and was confirmed at the protein, transcript and genomic levels. Downregulation of MBD4 expression using RNAi prevented the enhancement of gene correction efficacy obtained using the methyl-CpG-modified ssODN, demonstrating the specificity of the repair mechanism being recruited. Our data demonstrate that efficient manipulation of genomic targets can be achieved and controlled by the type of ssODN used and by modulation of the repair mechanism involved in the correction process. This new generation of ssODNs represents an important technological advance that is likely to have an impact on multiple applications, especially for gene therapy where permanent correction of the genetic defect has clear advantages over viral and other nonviral approaches currently being tested. PMID:19854937

  5. A review of statistical methods for preprocessing oligonucleotide microarrays. (United States)

    Wu, Zhijin


    Microarrays have become an indispensable tool in biomedical research. This powerful technology not only makes it possible to quantify a large number of nucleic acid molecules simultaneously, but also produces data with many sources of noise. A number of preprocessing steps are therefore necessary to convert the raw data, usually in the form of hybridisation images, to measures of biological meaning that can be used in further statistical analysis. Preprocessing of oligonucleotide arrays includes image processing, background adjustment, data normalisation/transformation and sometimes summarisation when multiple probes are used to target one genomic unit. In this article, we review the issues encountered in each preprocessing step and introduce the statistical models and methods in preprocessing.

  6. Antisense oligonucleotide targeting midkine suppresses in vivo angiogenesis

    Institute of Scientific and Technical Information of China (English)

    Li-Cheng Dai; Xiang Wang; Xing Yao; Yong-Liang Lu; Jin-Liang Ping; Jian-Fang He


    AIM: To evaluate the effect of antisense oligonucleotide targeting midkine (MK-AS) on angiogenesis in chick chorioallantoic membrane (CAM) andin situ human hepatocellular carcinoma (HCC).METHODS: An in situ human hepatocellular carcinoma (HCC) model and CAM assay were used in this experiment. The effect of MK-AS on angiogenesis was evaluated by cell proliferation assay and hematoxylineosin (HE) staining.RESULTS: MK-AS significantly inhibited human umbilical vein endothelial cells (HUVEC) and in situ human HCC growth. At the same time, MK-AS suppressed the angiogenesis both in human hepatocellular carcinoma cell line (HEPG2)-induced CAM and in situ human HCC tissues.CONCLUSION: MK-AS is an effective antiangiogenesis agent in vivo.

  7. Empirical evaluation of oligonucleotide probe selection for DNA microarrays.

    Directory of Open Access Journals (Sweden)

    Jennifer G Mulle

    Full Text Available DNA-based microarrays are increasingly central to biomedical research. Selecting oligonucleotide sequences that will behave consistently across experiments is essential to the design, production and performance of DNA microarrays. Here our aim was to improve on probe design parameters by empirically and systematically evaluating probe performance in a multivariate context. We used experimental data from 19 array CGH hybridizations to assess the probe performance of 385,474 probes tiled in the Duchenne muscular dystrophy (DMD region of the X chromosome. Our results demonstrate that probe melting temperature, single nucleotide polymorphisms (SNPs, and homocytosine motifs all have a strong effect on probe behavior. These findings, when incorporated into future microarray probe selection algorithms, may improve microarray performance for a wide variety of applications.

  8. Empirical evaluation of oligonucleotide probe selection for DNA microarrays. (United States)

    Mulle, Jennifer G; Patel, Viren C; Warren, Stephen T; Hegde, Madhuri R; Cutler, David J; Zwick, Michael E


    DNA-based microarrays are increasingly central to biomedical research. Selecting oligonucleotide sequences that will behave consistently across experiments is essential to the design, production and performance of DNA microarrays. Here our aim was to improve on probe design parameters by empirically and systematically evaluating probe performance in a multivariate context. We used experimental data from 19 array CGH hybridizations to assess the probe performance of 385,474 probes tiled in the Duchenne muscular dystrophy (DMD) region of the X chromosome. Our results demonstrate that probe melting temperature, single nucleotide polymorphisms (SNPs), and homocytosine motifs all have a strong effect on probe behavior. These findings, when incorporated into future microarray probe selection algorithms, may improve microarray performance for a wide variety of applications.

  9. Stem Cell Basics (United States)

    ... Tips Info Center Research Topics Federal Policy Glossary Stem Cell Information General Information Clinical Trials Funding Information Current ... Basics » Stem Cell Basics I. Back to top Stem Cell Basics I. Introduction: What are stem cells, and ...

  10. Basics of SCI Rehabilitation (United States)

    ... Donate Experts \\ The Basics of Spinal Cord Injury Rehabilitation Topics Adult Injuries Spinal Cord Injury 101 Spinal ... Injury 101 The Basics of Spinal Cord Injury Rehabilitation The Basics of Spinal Cord Injury Rehabilitation Preventing ...

  11. New Intelligent Method of Forecasting Control for Fishing Boat Security in the Coastal Area of Fujian

    Directory of Open Access Journals (Sweden)

    Wei He


    Full Text Available To ensure the fishing boat security in the coastal area of Fujian, it is imperative to develop efficient forecasting control system to manage the boats operation in a safe and orderly process. The core issue of establishing the forecasting control system is to predict the relationship of the real time traffic flow and the boat security condition. However, literature review shows that limited reports have addressed on this problem. Hence, a new intelligent forecasting control method base on the Chaos-Particle Swarm Optimization (PSO and Fuzzy Neural Network (FNN is proposed for the short time traffic flow prediction in this paper. The Empirical Mode Decomposition (EMD was first used to denoise the original ship traffic flow observation and then the Chaos-PSO-FNN was applied to the forecasting of the ship traffic flow and hence established the relationship of the real time traffic flow and the boat security condition. The advantage of the proposed approach is that the Chaos-PSO is employed to optimize the FNN parameters to overcome the premature problem of the FNN. As a result, the forecasting control performance is enhanced greatly. In the experiment analysis, the fishing boat traffic information provided by the Fujian marine bureau has been used to evaluate the newly proposed method. The analysis results demonstrate that the proposed Chaos-PSO-FNN method can extract distinct features of the traffic data and the prediction rate is beyond 93.7%. In addition, it found that the fishing boats is supposed to be safe when their travel time avoids the rush time of the traffic flow. This result agrees well with the real data. Thus, the new intelligent forecasting control method for fishing boat security can be used in practice.

  12. Direct microcontact printing of oligonucleotides for biochip applications

    Directory of Open Access Journals (Sweden)

    Trévisiol E


    Full Text Available Abstract Background A critical step in the fabrication of biochips is the controlled placement of probes molecules on solid surfaces. This is currently performed by sequential deposition of probes on a target surface with split or solid pins. In this article, we present a cost-effective procedure namely microcontact printing using stamps, for a parallel deposition of probes applicable for manufacturing biochips. Results Contrary to a previous work, we showed that the stamps tailored with an elastomeric poly(dimethylsiloxane material did not require any surface modification to be able to adsorb oligonucleotides or PCR products. The adsorbed DNA molecules are subsequently printed efficiently on a target surface with high sub-micron resolution. Secondly, we showed that successive stamping is characterized by an exponential decay of the amount of transferred DNA molecules to the surface up the 4th print, then followed by a second regime of transfer that was dependent on the contact time and which resulted in reduced quality of the features. Thus, while consecutive stamping was possible, this procedure turned out to be less reproducible and more time consuming than simply re-inking the stamps between each print. Thirdly, we showed that the hybridization signals on arrays made by microcontact printing were 5 to 10-times higher than those made by conventional spotting methods. Finally, we demonstrated the validity of this microcontact printing method in manufacturing oligonucleotides arrays for mutations recognition in a yeast gene. Conclusion The microcontact printing can be considered as a new potential technology platform to pattern DNA microarrays that may have significant advantages over the conventional spotting technologies as it is easy to implement, it uses low cost material to make the stamp, and the arrays made by this technology are 10-times more sensitive in term of hybridization signals than those manufactured by conventional spotting

  13. Design and development of thermolytic DNA oligonucleotide prodrugs. (United States)

    Grajkowski, Andrzej; Pedras-Vasconcelos, Joao; Ausín, Cristina; Verthelyi, Daniela; Beaucage, Serge L


    Deoxyribonucleoside phosphoramidites functionalized with the thermolytic 2-(N-formyl-N-methyl)aminoethyl group for phosphorus protection (1a-d) have been prepared and employed in the solid-phase synthesis of CpG ODN fma1555. Given that this modified oligonucleotide can be converted to the immunomodulatory CpG ODN 1555 under neutral conditions at 37 degrees C, its biologic activity was demonstrated in vivo by studies showing that intraperitoneal administration of CpG ODN fma1555 in mice resulted in the activation of cytokine-secreting splenocytes. Furthermore, administration of CpG ODN fma1555 to mice that were challenged intradermally in the ear with live L. major metacyclic promastigotes, reduced the severity of Leishmania skin lesions over time to an extent similar to that obtained with CpG ODN 1555. In another infectious model experiment, CpG ODN fma1555 protected newborn mice from death (65% survival) when administered 3 days before infection with the aggressive Tacaribe (TCRV) virus. A comparable immunoprotection was obtained by treatment of TCRV-infected mice with CpG ODN 1555 administered on the same day of infection (45% survival). However, when TCRV-infected mice were treated with CpG ODN fma1555 on the day of infection, they died as a consequence of the relatively slow conversion of the oligonucleotide prodrug to the bioactive CpG ODN 1555. Co-administration of both CpG ODN 1555 and CpG ODN fma1555 to mice 3 days prior to TCRV infection or on the day of infection provided protection from death (45-65% survival) and thus widened the immunoprotection window against TCRV-infection.

  14. Combining gene expression data from different generations of oligonucleotide arrays

    Directory of Open Access Journals (Sweden)

    Kong Sek


    Full Text Available Abstract Background One of the important challenges in microarray analysis is to take full advantage of previously accumulated data, both from one's own laboratory and from public repositories. Through a comparative analysis on a variety of datasets, a more comprehensive view of the underlying mechanism or structure can be obtained. However, as we discover in this work, continual changes in genomic sequence annotations and probe design criteria make it difficult to compare gene expression data even from different generations of the same microarray platform. Results We first describe the extent of discordance between the results derived from two generations of Affymetrix oligonucleotide arrays, as revealed in cluster analysis and in identification of differentially expressed genes. We then propose a method for increasing comparability. The dataset we use consists of a set of 14 human muscle biopsy samples from patients with inflammatory myopathies that were hybridized on both HG-U95Av2 and HG-U133A human arrays. We find that the use of the probe set matching table for comparative analysis provided by Affymetrix produces better results than matching by UniGene or LocusLink identifiers but still remains inadequate. Rescaling of expression values for each gene across samples and data filtering by expression values enhance comparability but only for few specific analyses. As a generic method for improving comparability, we select a subset of probes with overlapping sequence segments in the two array types and recalculate expression values based only on the selected probes. We show that this filtering of probes significantly improves the comparability while retaining a sufficient number of probe sets for further analysis. Conclusions Compatibility between high-density oligonucleotide arrays is significantly affected by probe-level sequence information. With a careful filtering of the probes based on their sequence overlaps, data from different

  15. The May 2015 boat crisis: the Rohingya in Aceh

    Directory of Open Access Journals (Sweden)

    Graham Thom


    Full Text Available Abstract In May 2015, Indonesia permitted the disembarkation of approximately 1,800 people who had been stranded at sea. The discovery of mass graves in Thailand’s Sadao district, on the border with Malaysia, led to a crack down on people smugglers by the Thai and Malaysian authorities. This saw thousands of Rohingya left stranded at sea as smugglers abandoned their human cargo. While initially pushed back by the Indonesian navy, after being rescued by Indonesian fishermen the Rohingya, on the three boats rescued, were permitted to stay in Indonesia’s Aceh province and seek protection. As part of an Amnesty International research team, conducting research in Aceh in August 2015, our aim was to document the reasons driving the Rohingya to leave Myanmar and Bangladesh, the human rights abuses they experienced at sea and the conditions they are experiencing in Aceh. The subsequent Amnesty International report primarily documented the abuses suffered by those on board the smuggling/trafficking vessels. It also highlighted a number of issues now facing those allowed to remain in Aceh (Amnesty International, 2015. This article will further examine the human rights situation facing the Rohingya in Aceh, why they have been treated differently from previous Rohingya groups who have arrived in Aceh and the ongoing sustainability of the current model. Significantly, the majority of those that arrived have subsequently escaped from the “sites”[1], having again engaged smugglers to reach Malaysia. Given that a process already exists for refugees arriving in Indonesia, the differing treatment received by the Rohingya arriving in May 2015 raises a number of questions. This paper will conclude that the current situation is not sustainable and the treatment of refugees in Aceh should be included in a broader national approach commensurate to the treatment of refugees and asylum seekers throughout Indonesia. [1] Rather than detention centres or camps, the

  16. Tank tests of a family of flying-boat hulls (United States)

    Shoemaker, James M; Parkinson, John B


    This report presents towing tests made in the N.A.C.A. tank of a parent form and five variations of a flying-boat hull. The beams of two of the derived forms were made the same as that of the parent and the lengths changed by increasing and decreasing the spacing of stations. The lengths of the two others of the derived forms were made the same as that of the parent while the beams were changed by increasing and decreasing the spacing of buttocks, all other widths being changed in proportion. The remaining derived form has the same length and beam as the parent, but the lines of the forebody were altered to give a planing bottom with no longitudinal curvature forward of the step. The test data were analyzed to determine the minimum resistance and the angle at which it occurs for all speeds and loads. The results of this analysis are given in the form of non dimensional curves for each model. The effect of variation in over-all size, as indicated by a "complete" test on any given hull, is pointed out. The effect of changing length alone by the spacing of buttocks, as well as the effects of the changes in length-beam ratio and longitudinal curvature that result from these operations are discussed. The difficulties encountered in interpreting test results of systematic families derived by the method used are emphasized. Further studies are suggested in which changes in the variable under consideration would not be obscured by secondary changes in other important variables.

  17. Boats, Borders and the Geo-Imaginaries of the South

    Directory of Open Access Journals (Sweden)

    Francis Maravillas


    Full Text Available This paper activates a mode of spatial inquiry into Australia’s identity through an analysis of a number of frames through which the passage and interdiction of boats off the coast of the nation may be viewed. In particular, I explore the way in which Australia’s paradoxical geographical location as South of both the West and Asia play a key role in affixing the horizon within which a conception of the nation and its relationship with the world was – and continues to be – defined and shaped. Moreover, I not only critically probe the constitutive fears and anxieties that underlie bounded conceptions of the trope of the South, but also to examine how such a trope can articulate itself as a site of exchange and negotiation, a distinctive borderland that engenders new cartographies of difference and belonging in an increasingly globalised and interconnected world. I show how these frames overlap and converge on the wider questions of space, place and identity at the very moment when the process of globalisation and migration has done so much to shake any certainties about Australia’s identity as a geographically distinct and spatially bounded nation-state. In so doing, they represent crucial sites for articulating and enacting a transcultural politics of mobility and spatiality that attends to the ways in which the trope of the South may been imagined not as a sphere of containment or an enclaved territory, but as an evolving cartography, the shifting outlines of which opens up new horizons of possibility for rethinking the spatial and temporal coordinates of Australia in a globalizing world.

  18. Release of DNA oligonucleotides and their conjugates from controlled-pore glass under thermolytic conditions. (United States)

    Grajkowski, Andrzej; Cieślak, Jacek; Norris, Scott; Freedberg, Darón I; Kauffman, Jon S; Duff, Robert J; Beaucage, Serge L


    The sequential functionalization of long-chain alkylamine controlled-pore glass (CPG) with a 3-hydroxypropyl-(2-cyanoethyl)thiophosphoryl linker and a dinucleoside phosphorotetrazolide leads to a uniquely engineered support for solid-phase synthesis. Unlike conventional succinylated-CPG supports, this support is designed to allow oligonucleotide deprotection and elimination of deprotection side-products to proceed without release of the oligonucleotide. When needed, the DNA oligonucleotide can be thermolytically released in 2 hr under essentially neutral conditions. The modified CPG support has been successfully employed in the synthesis of both native and fully phosphorothioated DNA 20-mers. On the basis of reversed-phase HPLC and electrophoretic analyses, the purity of the released oligonucleotides is comparable to that of identical oligonucleotides synthesized from succinylated-CPG supports, in terms of both shorter-than-full-length oligonucleotide contaminants and overall yields. The detailed preparation of DNA oligonucleotides conjugated with exemplary reporter or functional groups, either at the 3'-terminus or at both 3'- and 5'-termini, is also described.

  19. Polymerase-endonuclease amplification reaction (PEAR for large-scale enzymatic production of antisense oligonucleotides.

    Directory of Open Access Journals (Sweden)

    Xiaolong Wang

    Full Text Available Antisense oligonucleotides targeting microRNAs or their mRNA targets prove to be powerful tools for molecular biology research and may eventually emerge as new therapeutic agents. Synthetic oligonucleotides are often contaminated with highly homologous failure sequences. Synthesis of a certain oligonucleotide is difficult to scale up because it requires expensive equipment, hazardous chemicals and a tedious purification process. Here we report a novel thermocyclic reaction, polymerase-endonuclease amplification reaction (PEAR, for the amplification of oligonucleotides. A target oligonucleotide and a tandem repeated antisense probe are subjected to repeated cycles of denaturing, annealing, elongation and cleaving, in which thermostable DNA polymerase elongation and strand slipping generate duplex tandem repeats, and thermostable endonuclease (PspGI cleavage releases monomeric duplex oligonucleotides. Each round of PEAR achieves over 100-fold amplification. The product can be used in one more round of PEAR directly, and the process can be further repeated. In addition to avoiding dangerous materials and improved product purity, this reaction is easy to scale up and amenable to full automation. PEAR has the potential to be a useful tool for large-scale production of antisense oligonucleotide drugs.

  20. Wave Motion Compensation Scheme and Its Model Tests for the Salvage of An Ancient Sunken Boat

    Institute of Scientific and Technical Information of China (English)

    YE Jia-wei; CHEN Yuan-ming; WANG Dong-jiao; LIU Yue-qin; SONG Xin; HUANG Yuan-tian


    The application of the vertical hoisting jack and wave motion compensation techniques to the salvage of an ancient sunken boat is introduced. The boat is wooden, loaded with cultural relics. It has been immersed at the bottom of the South China Sea for more than 800 years. In order to protect the structure of the boat and the cultural relics inside to the largest extent, an open caisson is used to hold the sunken boat and the silts around before they are raised from the seabed all together as a whole. In the paper, first, the seakeeping model test of the system of the salvage barge and the open caisson is done to determine some important wave response parameters. And then a further experimental study of the application of the vertical hoisting jack and wave motion compensation scheme to the salvage of the sunken boat is carried out. In the model tests, the techniques of the integrative mechanic-electronic-hydraulic control, wave motion forecast and wave motion compensation are used to minimize the heave motion of the open caisson. The results of the model tests show that the heave motion of the open caisson can be reduced effectively by the use of the present method.

  1. Design of a semi-autonomous boat for measurements of coastal sedimentation and erosion (United States)

    Smith, D.; Cross, L.; Rivet, J.; Hall, S.


    Measurement of sediment deposition and erosion in coastal areas is a challenge due to soft shifting sediments, but is critical to assessing loss or restoration of coastal sediments and wetlands. The aim of this project was to design and construct a semi-autonomous boat with water depth measuring capabilities. It was intended to map the depth of coastal wetlands to determine erosion rates and assess coastal restoration effects. Depth-measuring equipment was incorporated into an autonomous pontoon boat powered by solar panels. The propulsion system consisted of two paddlewheels and two-way motors to allow movement and positioning for measurements. Modifications included a lightweight, hard coating on the pontoons and powder-coating the frame to extend their usable life. A microcontroller controlled the boat and captured depth data from sensors and location data with a GPS system. The depth measuring system consisted of a pulley and counter system that completed each measurement in less than 45 seconds. This allowed the boat to take approximately 400 measurements per day. Net accuracy was approximately 3 cm in the tested configuration. The boat can continually measure the depth of specified areas in the wetlands; with this data, the change in depth can be monitored to see the effects of restoration projects.

  2. Chimeric RNA Oligonucleotides with Triazole and Phosphate Linkages: Synthesis and RNA Interference. (United States)

    Fujino, Tomoko; Kogashi, Kanako; Okada, Koudai; Mattarella, Martin; Suzuki, Takeru; Yasumoto, Kenichi; Sogawa, Kazuhiro; Isobe, Hiroyuki


    Chimeric RNA oligonucleotides with an artificial triazole linker were synthesized using solution-phase click chemistry and solid-phase automated synthesis. Scalable synthesis methods for jointing units for the chimeric structure have been developed, and after click-coupling of the jointing units with triazole linkers, a series of chimeric oligonucleotides was prepared by utilizing the well-established phosphoramidite method for the elongation. The series of chimeric 21-mer oligonucleotides that possessed the triazole linker at different strands and positions allowed for a screening study of the RNA interference to clarify the preference of the triazole modifications in small-interfering RNA molecules.

  3. Research and development of Ro-boat: an autonomous river cleaning robot (United States)

    Sinha, Aakash; Bhardwaj, Prashant; Vaibhav, Bipul; Mohommad, Noor


    Ro-Boat is an autonomous river cleaning intelligent robot incorporating mechanical design and computer vision algorithm to achieve autonomous river cleaning and provide a sustainable environment. Ro-boat is designed in a modular fashion with design details such as mechanical structural design, hydrodynamic design and vibrational analysis. It is incorporated with a stable mechanical system with air and water propulsion, robotic arms and solar energy source and it is proceed to become autonomous by using computer vision. Both "HSV Color Space" and "SURF" are proposed to use for measurements in Kalman Filter resulting in extremely robust pollutant tracking. The system has been tested with successful results in the Yamuna River in New Delhi. We foresee that a system of Ro-boats working autonomously 24x7 can clean a major river in a city on about six months time, which is unmatched by alternative methods of river cleaning.

  4. 19 CFR 10.36a - Vehicles, pleasure boats and aircraft brought in for repair or alteration. (United States)


    ... RATE, ETC. General Provisions Temporary Importations Under Bond § 10.36a Vehicles, pleasure boats and... shall be without surety or cash deposit except as provided by this paragraph and paragraph (d) of this... supported by bond with surety or cash deposit in lieu of surety. (b) Each vehicle, pleasure boat,...

  5. 77 FR 15006 - Special Local Regulations; Third Annual Space Coast Super Boat Grand Prix, Atlantic Ocean, Cocoa... (United States)


    ... SECURITY Coast Guard 33 CFR Part 100 RIN 1625-AA08 Special Local Regulations; Third Annual Space Coast... of the Atlantic Ocean east of Cocoa Beach, Florida during the Third Annual Space Coast Super Boat Grand Prix, a series of high-speed boat races. The event is scheduled to take place on Sunday, May...

  6. 33 CFR 150.505 - When must the operator service and examine lifeboat and rescue boat release gear? (United States)


    ... inspection by personnel trained in examining the gear. Inflatable Lifesaving Appliances ... and examine lifeboat and rescue boat release gear? 150.505 Section 150.505 Navigation and Navigable... and examine lifeboat and rescue boat release gear? (a) The operator must service lifeboat and...

  7. 78 FR 60698 - Safety Zone, Lucas Oil Drag Boat Racing Series; Thompson Bay, Lake Havasu City, AZ. (United States)


    ... SECURITY Coast Guard 33 CFR Part 165 RIN 1625-AA00 Safety Zone, Lucas Oil Drag Boat Racing Series; Thompson... Guard is establishing a temporary safety zone within the navigable waters of Thompson Bay in Lake Havasu... Thompson Bay, Lake Havasu, AZ for The Lucas Oil Drag Boat Racing Series. This safety zone is necessary...

  8. 26 CFR 31.3401(a)(17)-1 - Remuneration for services performed on a boat engaged in catching fish. (United States)


    ... 26 Internal Revenue 15 2010-04-01 2010-04-01 false Remuneration for services performed on a boat... Remuneration for services performed on a boat engaged in catching fish. (a) Remuneration for services performed... receive, any cash remuneration, other than remuneration that is described in subparagraph (1) of...

  9. 40 CFR Table 1 to Subpart Vvvv of... - Compliance Dates for New and Existing Boat Manufacturing Facilities (United States)


    ... Boat Manufacturing Facilities 1 Table 1 to Subpart VVVV of Part 63 Protection of Environment... Manufacturing Pt. 63, Subpt. VVVV, Table 1 Table 1 to Subpart VVVV of Part 63—Compliance Dates for New and Existing Boat Manufacturing Facilities As specified in § 63.5695, you must comply by the dates in...

  10. Tank tests of a model of a flying-boat hull with a fluted bottom (United States)

    Dawson, John R


    A 1/5-scale model of a flying-boat hull having flutes in the bottom both forward and aft of the step (NACA model 19) was tested to determine its water performance. The model was also tested after the successive removal of the flutes on the afterbody and forebody. The results from these tests are compared with those from tests of a model of the hull of the Navy PN-8 flying boat and it is concluded that the fluted-bottom model and its modifications are inferior to the model of the PN-8.


    Directory of Open Access Journals (Sweden)



    Full Text Available In this paper Gain scheduling PD controller and Adaptive PD controller are adopted as autopilot systems for a boat model. The control design is based on Nomoto’s first order mathematical model. Adaptive controller is based on model reference adaptive controller theory. Similar test runs are conducted on a model boat with the above controller algorithms. The effectiveness of the two controllers are compared numerically and graphically. The results show that both controllers are well for reaching the desired heading angle. The adaptive controller has some notable advantage over gain scheduling controller.

  12. Oligonucleotide-based biosensors for in vitro diagnostics and environmental hazard detection. (United States)

    Jung, Il Young; Lee, Eun Hee; Suh, Ah Young; Lee, Seung Jin; Lee, Hyukjin


    Oligonucleotide-based biosensors have drawn much attention because of their broad applications in in vitro diagnostics and environmental hazard detection. They are particularly of interest to many researchers because of their high specificity as well as excellent sensitivity. Recently, oligonucleotide-based biosensors have been used to achieve not only genetic detection of targets but also the detection of small molecules, peptides, and proteins. This has further broadened the applications of these sensors in the medical and health care industry. In this review, we highlight various examples of oligonucleotide-based biosensors for the detection of diseases, drugs, and environmentally hazardous chemicals. Each example is provided with detailed schematics of the detection mechanism in addition to the supporting experimental results. Furthermore, future perspectives and new challenges in oligonucleotide-based biosensors are discussed.

  13. Exploiting Protected Maleimides to Modify Oligonucleotides, Peptides and Peptide Nucleic Acids

    Directory of Open Access Journals (Sweden)

    Clément Paris


    Full Text Available This manuscript reviews the possibilities offered by 2,5-dimethylfuran-protected maleimides. Suitably derivatized building blocks incorporating the exo Diels-Alder cycloadduct can be introduced at any position of oligonucleotides, peptide nucleic acids, peptides and peptoids, making use of standard solid-phase procedures. Maleimide deprotection takes place upon heating, which can be followed by either Michael-type or Diels-Alder click conjugation reactions. However, the one-pot procedure in which maleimide deprotection and conjugation are simultaneously carried out provides the target conjugate more quickly and, more importantly, in better yield. This procedure is compatible with conjugates involving oligonucleotides, peptides and peptide nucleic acids. A variety of cyclic peptides and oligonucleotides can be obtained from peptide and oligonucleotide precursors incorporating protected maleimides and thiols.

  14. Nucleoside, nucleotide and oligonucleotide based amphiphiles: a successful marriage of nucleic acids with lipids. (United States)

    Gissot, Arnaud; Camplo, Michel; Grinstaff, Mark W; Barthélémy, Philippe


    Amphiphilic molecules based on nucleosides, nucleotides and oligonucleotides are finding more and more biotechnological applications. This Perspective highlights their synthesis, supramolecular organization as well as their applications in the field of biotechnology.

  15. Integrated synoptic surveys of the hydrodynamics and water-quality distributions in two Lake Michigan rivermouth mixing zones using an autonomous underwater vehicle and a manned boat (United States)

    Jackson, P. Ryan; Reneau, Paul C.


    The U.S. Geological Survey (USGS), in cooperation with the National Monitoring Network for U.S. Coastal Waters and Tributaries, launched a pilot project in 2010 to determine the value of integrated synoptic surveys of rivermouths using autonomous underwater vehicle technology in response to a call for rivermouth research, which includes study domains that envelop both the fluvial and lacustrine boundaries of the rivermouth mixing zone. The pilot project was implemented at two Lake Michigan rivermouths with largely different scales, hydrodynamics, and settings, but employing primarily the same survey techniques and methods. The Milwaukee River Estuary Area of Concern (AOC) survey included measurements in the lower 2 to 3 miles of the Milwaukee, Menomonee, and Kinnickinnic Rivers and inner and outer Milwaukee Harbor. This estuary is situated in downtown Milwaukee, Wisconsin, and is the most populated basin that flows directly into Lake Michigan. In contrast, the Manitowoc rivermouth has a relatively small harbor separating the rivermouth from Lake Michigan, and the Manitowoc River Watershed is primarily agricultural. Both the Milwaukee and Manitowoc rivermouths are unregulated and allow free exchange of water with Lake Michigan. This pilot study of the Milwaukee River Estuary and Manitowoc rivermouth using an autonomous underwater vehicle (AUV) paired with a manned survey boat resulted in high spatial and temporal resolution datasets of basic water-quality parameter distributions and hydrodynamics. The AUV performed well in these environments and was found primarily well-suited for harbor and nearshore surveys of three-dimensional water-quality distributions. Both case studies revealed that the use of a manned boat equipped with an acoustic Doppler current profiler (ADCP) and multiparameter sonde (and an optional flow-through water-quality sampling system) was the best option for riverine surveys. To ensure that the most accurate and highest resolution velocity data

  16. Kinetic Hairpin Oligonucleotide Blockers for Selective Amplification of Rare Mutations (United States)

    Jia, Yanwei; Sanchez, J. Aquiles; Wangh, Lawrence J.


    Detection of rare mutant alleles in an excess of wild type alleles is increasingly important in cancer diagnosis. Several methods for selective amplification of a mutant allele via the polymerase chain reaction (PCR) have been reported, but each of these methods has its own limitations. A common problem is that Taq DNA polymerase errors early during amplification generate false positive mutations which also accumulate exponentially. In this paper, we described a novel method using hairpin oligonucleotide blockers that can selectively inhibit the amplification of wild type DNA during LATE-PCR amplification. LATE-PCR generates double-stranded DNA exponentially followed by linear amplification of single-stranded DNA. The efficiency of the blocker is optimized by adjusting the LATE-PCR temperature cycling profile. We also demonstrate that it is possible to minimize false positive signals caused by Taq DNA polymerase errors by using a mismatched excess primer plus a modified PCR profile to preferentially enrich for mutant target sequences prior to the start of the exponential phase of LATE-PCR amplification. In combination these procedures permit amplification of specific KRAS mutations in the presence of more than 10,000 fold excess of wild type DNA without false positive signals. PMID:25082368

  17. Advantages of ion-exchange chromatography for oligonucleotide analysis. (United States)

    Cook, Ken; Thayer, Jim


    The rapid development of therapeutic oligonucleotides (ONs) has created a need for in-depth characterization of ONs, beyond previous requirements. The natural migration to LC-MS requires the use of chromatography with MS-compatible eluents to introduce the large, highly charged biopolymers into the mass spectrometer. Most frequently this employs ion-pair reversed-phase liquid chromatography, which may leave gaps in the characterization, but these can be filled with the use of high-resolution ion-exchange chromatography. Several classes of isobaric isomers are among the impurities that will require further separation prior to MS analysis. This review shows how the use of ion exchange as an additional orthogonal analytical method can be used as standalone or interfaced with MS to achieve the highest possible analytical coverage in the characterization and quantification of impurities present in single- and double-stranded ON formulations. Some of these techniques have been in use for some time and the importance of others is just being recognized.

  18. Antineoplastic effect of decoy oligonucleotide derived from MGMT enhancer.

    Directory of Open Access Journals (Sweden)

    Tamar Canello

    Full Text Available Silencing of O(6-methylguanine-DNA-methyltransferase (MGMT in tumors, mainly through promoter methylation, correlates with a better therapeutic response and with increased survival. Therefore, it is conceivable to consider MGMT as a potential therapeutic target for the treatment of cancers. Our previous results demonstrated the pivotal role of NF-kappaB in MGMT expression, mediated mainly through p65/NF-kappaB homodimers. Here we show that the non-canonical NF-KappaB motif (MGMT-kappaB1 within MGMT enhancer is probably the major inducer of MGMT expression following NF-kappaB activation. Thus, in an attempt to attenuate the transcription activity of MGMT in tumors we designed locked nucleic acids (LNA modified decoy oligonucleotides corresponding to the specific sequence of MGMT-kappaB1 (MGMT-kB1-LODN. Following confirmation of the ability of MGMT-kB1-LODN to interfere with the binding of p65/NF-kappaB to the NF-KappaB motif within MGMT enhancer, the efficacy of the decoy was studied in-vitro and in-vivo. The results of these experiments show that the decoy MGMT-kB1-LODN have a substantial antineoplastic effect when used either in combination with temozolomide or as monotherapy. Our results suggest that MGMT-kB1-LODN may provide a novel strategy for cancer therapy.

  19. Integrated Microfluidic Isolation of Aptamers Using Electrophoretic Oligonucleotide Manipulation (United States)

    Kim, Jinho; Olsen, Timothy R.; Zhu, Jing; Hilton, John P.; Yang, Kyung-Ae; Pei, Renjun; Stojanovic, Milan N.; Lin, Qiao


    We present a microfluidic approach to integrated isolation of DNA aptamers via systematic evolution of ligands by exponential enrichment (SELEX). The approach employs a microbead-based protocol for the processes of affinity selection and amplification of target-binding oligonucleotides, and an electrophoretic DNA manipulation scheme for the coupling of these processes, which are required to occur in different buffers. This achieves the full microfluidic integration of SELEX, thereby enabling highly efficient isolation of aptamers in drastically reduced times and with minimized consumption of biological material. The approach as such also offers broad target applicability by allowing selection of aptamers with respect to targets that are either surface-immobilized or solution-borne, potentially allowing aptamers to be developed as readily available affinity reagents for a wide range of targets. We demonstrate the utility of this approach on two different procedures, respectively for isolating aptamers against a surface-immobilized protein (immunoglobulin E) and a solution-phase small molecule (bisboronic acid in the presence of glucose). In both cases aptamer candidates were isolated in three rounds of SELEX within a total process time of approximately 10 hours.

  20. The development of bioactive triple helix-forming oligonucleotides. (United States)

    Seidman, Michael M; Puri, Nitin; Majumdar, Alokes; Cuenoud, Bernard; Miller, Paul S; Alam, Rowshon


    We are developing triple helix-forming oligonucleotides (TFOs) as gene targeting reagents in mammalian cells. We have described psoralen-conjugated TFOs containing 2'-O-methyl (2'OMe) and 2'-O-aminoethoxy (AE) ribose substitutions. TFOs with a cluster of 3-4 AE residues, with all other sugars as 2'OMe, were bioactive in a gene knockout assay in mammalian cells. In contrast, TFOs with one or two clustered, or three dispersed, AE residues were inactive. Thermal stability analysis of the triplexes indicated that there were only incremental differences between the active and inactive TFOs. However the active and inactive TFOs could be distinguished by their association kinetics. The bioactive TFOs showed markedly greater on-rates than the inactive TFOs. It appears that the on-rate is a better predictor of TFO bioactivity than thermal stability. Our data are consistent with a model in which a cluster of 3-4 AE residues stabilizes the nucleation event that precedes formation of a complete triplex. It is likely that triplexes in cells are much less stable than triplexes in vitro probably as a result of elution by chromatin-associated translocases and helicases. Consequently the biologic assay will favor TFOs that can bind and rebind genomic targets quickly.

  1. Effect of oligonucleotide primers in determining viral variability within hosts

    Directory of Open Access Journals (Sweden)

    Moya Andrés


    Full Text Available Abstract Background Genetic variability in viral populations is usually estimated by means of polymerase chain reaction (PCR based methods in which the relative abundance of each amplicon is assumed to be proportional to the frequency of the corresponding template in the initial sample. Although bias in template-to-product ratios has been described before, its relevance in describing viral genetic variability at the intrapatient level has not been fully assessed yet. Results To investigate the role of oligonucleotide design in estimating viral variability within hosts, genetic diversity in hepatitis C virus (HCV populations from eight infected patients was characterised by two parallel PCR amplifications performed with two slightly different sets of primers, followed by cloning and sequencing (mean = 89 cloned sequences per patient. Population genetics analyses of viral populations recovered by pairs of amplifications revealed that in seven patients statistically significant differences were detected between populations sampled with different set of primers. Conclusions Genetic variability analyses demonstrates that PCR selection due to the choice of primers, differing in their degeneracy degree at some nucleotide positions, can eclipse totally or partially viral variants, hence yielding significant different estimates of viral variability within a single patient and therefore eventually producing quite different qualitative and quantitative descriptions of viral populations within each host.

  2. Advancements of antisense oligonucleotides in treatment of breast cancer

    Institute of Scientific and Technical Information of China (English)

    YANGShuan-Ping; SONGSan-Tai; 等


    Breast cancer is one kind of multi-gene related malignancy.Overexpression of some oncogenes such as HER-2(c-erbB-2,Neu),bcl-2/bcl-xL,protein kinase A(PKA),and transferrin receptor gene(TfR gene),etc significantly affect the prognosis of breast cancer.It was shown that specific suppression of the overexpressed genes above resulted in the improvement of the therapy of breast cancer.Antisense of useful tools for inhibiting the overexpression of specific oncogenes,was involved in the therapy of breast cancer in recent years. Data indicated that antisense oligonucleotides(ON)could inhibit specially the expression of the target genes on mRNA or protein levels in most of cases;some ON candidates showed encouraging therapeutic effects in vitro and in vivo on breast cancer cell lines or xenografts.Furthermore,the combination use of the antisense ON and normal chemotherapeutic agents indicated synergistic antitumor effects,which was probably the best utilization of antisense ON in the treatment of breast cancer.

  3. Antineoplastic effect of decoy oligonucleotide derived from MGMT enhancer. (United States)

    Canello, Tamar; Ovadia, Haim; Refael, Miri; Zrihan, Daniel; Siegal, Tali; Lavon, Iris


    Silencing of O(6)-methylguanine-DNA-methyltransferase (MGMT) in tumors, mainly through promoter methylation, correlates with a better therapeutic response and with increased survival. Therefore, it is conceivable to consider MGMT as a potential therapeutic target for the treatment of cancers. Our previous results demonstrated the pivotal role of NF-kappaB in MGMT expression, mediated mainly through p65/NF-kappaB homodimers. Here we show that the non-canonical NF-KappaB motif (MGMT-kappaB1) within MGMT enhancer is probably the major inducer of MGMT expression following NF-kappaB activation. Thus, in an attempt to attenuate the transcription activity of MGMT in tumors we designed locked nucleic acids (LNA) modified decoy oligonucleotides corresponding to the specific sequence of MGMT-kappaB1 (MGMT-kB1-LODN). Following confirmation of the ability of MGMT-kB1-LODN to interfere with the binding of p65/NF-kappaB to the NF-KappaB motif within MGMT enhancer, the efficacy of the decoy was studied in-vitro and in-vivo. The results of these experiments show that the decoy MGMT-kB1-LODN have a substantial antineoplastic effect when used either in combination with temozolomide or as monotherapy. Our results suggest that MGMT-kB1-LODN may provide a novel strategy for cancer therapy.

  4. Nanoexplosive gene therapy using triplex-forming oligonucleotides

    Energy Technology Data Exchange (ETDEWEB)

    Oh, Eun Jung; Min, Hye Jung; Choe, Jae Gol; Park, Gil Hong; Kim, Meyoung Kon [College of Medicine, Korea Univ., Seoul (Korea, Republic of)


    Triplex forming oligonucleotides (TFO) labeled with Auger emitter could be ideal vehicles for delivering radiation energy to specific DNA sequences, and followed by double-stranded DNA breaks and subsequent inactivation of targeted genes. We designed TFOs targeting the selected DNA fragments (i.e., estrogen receptors and N-myc promoter) and labeled with {sup 125}I and {sup 111}In. Various Cancer cells, e.g., MCF-7 (breast adenocarcinoma), MCF-10A (immortalized breast cells), Jurkat (T-cell leukemia), ARO (thyroid cancer), SNU-449 (Colon Caner), and HL-60 (polymyelocytic leukemia), were prepared and treated with radiolabeled TFO for 24 h. After the incubation, subcellular fractions (i.e., cell nucleus, cytoplasm and cultured medium) were collected and measured radioactivity by a gamma scintillation counter, respectively. The mean value of % injected dose for each fraction was ranged as follows: nucleus, 4.4-20%; cytoplasm, 8.2-29%; and medium, 64-87%. Therefore, we speculated that TFO labeled with Auger emitter could be a next-generation therapeutic tool in nanoexplosive gene therapy.

  5. Respirable antisense oligonucleotides: a new drug class for respiratory disease

    Directory of Open Access Journals (Sweden)

    Tanaka Makoto


    Full Text Available Abstract Respirable antisense oligonucleotides (RASONs, which attenuate specific disease-associated mRNAs, represent a new class of respiratory therapeutics with considerable potential. RASONs overcome previous obstacles that have impeded the development of antisense therapeutics targeting diseases in other organ systems. RASONs are delivered directly to the target tissue via inhalation; their uptake seems to be enhanced by cationic properties inherent in pulmonary surfactant, and, because of the markedly different target properties of mRNA and proteins, they can have very long durations of effect compared with traditional drugs targeting the protein of the same gene. RASONs contain chemical modifications that decrease their degradation by cellular nucleases. However, total insensitivity to nucleases is probably not an optimal design criterion for RASONs, because moderate nuclease sensitivity can prevent their systemic delivery, decreasing the potential for systemic toxicity. EPI-2010 is a 21-mer phosphorothioate RASON that attenuates bronchoconstriction, inflammation and surfactant depletion in preclinical models of human asthma, has a duration of effect of seven days, and seems to undergo minimal systemic delivery.

  6. Sheath liquid effects in capillary high-performance liquid chromatography-electrospray mass spectrometry of oligonucleotides. (United States)

    Huber, C G; Krajete, A


    Fused-silica capillary columns of 200 microm inner diameter were packed with micropellicular, octadecylated, 2.3 microm poly(styrene-divinylbenzene) particles and applied to the separation of oligonucleotides by ion-pair reversed-phase high-performance liquid chromatography. Oligonucleotides were eluted at 50 degrees C with gradients of 3-13% acetonitrile in 50 mM triethylammonium bicarbonate. Addition of sheath liquid to the column effluent allowed the detection of oligonucleotides by electrospray ionization mass spectrometry using full-scan data acquisition with a detectability comparable to that obtained with UV detection. The signal-to-noise ratios with different sheath liquids increased in the order isopropanololigonucleotides longer than 20 nucleotide units whereas no significant effect was observed with shorter oligonucleotides. Organic acids and bases in the sheath liquid generally deteriorated the signal-to-noise ratios in the chromatograms and mass spectra mainly because of increased background noise. Only a few charge states were observed in the mass spectra of oligonucleotides because of charge state reduction due to the presence of carbonic acid in the eluent. With triethylammonium hydrogencarbonate as chromatographic eluent and acetonitrile as sheath liquid, very few cation adducts of oligonucleotides were observed in the mass spectra. However, the presence of small amounts of monopotassium adducts enabled the calculation of the charge state of multiply charged ions. With acetonitrile as sheath liquid, 710 amol of a 16-mer oligonucleotide were detected using selected ion monitoring data acquisition with a signal-to-noise ratio of 3:1. Finally, capillary ion-pair reversed-phase high-performance liquid chromatography-electrospray ionization mass spectrometry was

  7. Complexes of carbon nanotubes with oligonucleotides in thin Langmuir-Blodgett films to detect electrochemically hybridization (United States)

    Egorov, A. S.; Egorova, V. P.; Krylova, H. V.; Lipnevich, I. V.; Orekhovskaya, T. I.; Veligura, A. A.; Govorov, M. I.; Shulitsky, B. G.


    Self-assembled complexes consisting of thin multi-walled carbon nanotubes (MWCNTs) and DNA-oligonucleotides which are able to a cooperative binding to complementary oligonucleotides have been investigated. It was establised a high-performance charge transport in nanostructured Langmuir-Blodgett complexes thin MWCNTs/DNA. A method to electrochemically detect DNA hybridization on the self-organized structures has been proposed.

  8. Conceptual "Heat-Driven" approach to the synthesis of DNA oligonucleotides on microarrays. (United States)

    Grajkowski, A; Cieślak, J; Chmielewski, M K; Marchán, V; Phillips, L R; Wilk, A; Beaucage, S L


    The discovery of deoxyribonucleoside cyclic N-acylphosphoramidites, a novel class of phosphoramidite monomers for solid-phase oligonucleotide synthesis, has led to the development of a number of phosphate protecting groups that can be cleaved from DNA oligonucleotides under thermolytic neutral conditions. These include the 2-(N-formyl-N-methyl)aminoethyl, 4-oxopentyl, 3-(N-tert-butyl)carboxamido-1-propyl, 3-(2-pyridyl)-1-propyl, 2-[N-methyl-N-(2-pyridyl)]aminoethyl, and 4-methythiobutyl groups. When used for 5'-hydroxyl protection of nucleosides, the analogous 1-phenyl-2-[N-methyl-N-(2-pyridyl)]aminoethyloxycarbonyl group exhibited excellent thermolytic properties, which may permit an iterative "heat-driven" synthesis of DNA oligonucleotides on microarrays. In this regard, progress has been made toward the use of deoxyribonucleoside cyclic N-acylphosphoramidites in solid-phase oligonucleotide syntheses without nucleobase protection. Given that deoxyribonucleoside cyclic N-acylphosphoramidites produce oligonucleotides with heat-sensitive phosphate protecting groups, blocking the 5'-hydroxyl of these monomers with, for example, the thermolabile 1-phenyl-2-[N-methyl-N-(2-pyridyl)]aminoethyloxycarbonyl group may provide a convenient thermo-controlled method for the synthesis of oligonucleotides on microarrays.

  9. Strategies in the preparation of DNA oligonucleotide arrays for diagnostic applications. (United States)

    Beaucage, S L


    This report emphasizes the interfacial chemistry that is required to ensure proper attachment of oligonucleotides onto the surface of microarrays. For example, strategies for the covalent attachment of pre-synthesized oligonucleotides to glass slides, gold films, polyacrylamide gel pads, polypyrrole films, and optical fibers are surveyed in an attempt to better define the parameters for optimal formation and detection of DNA hybrids. These parameters include among others, the nature and length of the linkers attaching oligonucleotides to the arrays, and the surface density of oligonucleotides required for unhindered hybridization with DNA targets. Sensitive detection methods such as the use of light-scattering techniques, molecular beacons, surface plasmon resonance, attenuated total internal reflection-FTIR, and the evanescent field excitation of fluorescence from surface-bound fluorophores have been developed to study the kinetics and specificity of hybridization events. Finally, the synthesis of oligonucleotides directly on glass surfaces and polypropylene sheets has been investigated to enable DNA sequencing by hybridization and achieve oligonucleotide densities of ca. 10(6) sequences per cm(2) on DNA chips.

  10. Oligonucleotide chip, real-time PCR and sequencing for genotyping of hepatitis B virus

    Institute of Scientific and Technical Information of China (English)

    Yong-Zhong Wang; Guo-Xiang Wu; Li-Bo Luo; Min Chen; Li-Hua Ruan


    AIM: To compare the oligonucleotide chip, real-time PCR and sequencing for genotyping of hepatitis B virus in Chinese patients with chronic hepatitis B.METHODS: Mixture of samples with different genotypes and clinical serum samples from 126 chronic hepatitis B patients was tested for hepatitis B virus genotypes by oligonucleotide chip, real-time PCR and sequencing of PCR products, respectively. Clinical performances, time required and costs of the three assays were evaluated.RESULTS: Oligonucleotide chips and real-time PCR detected 1% and 0.1% genotypes, respectively, in mixed samples. Of the 126 clinical samples from patients with chronic hepatitis B, genotype B was detected in 41(33%), 41 (33%) and 45 (36%) samples, and genotype C in 76 (60%), 76 (60%) and 81 (64%) samples, by oligonucleotide chip, real-time PCR and sequencing,respectively. Oligonucleotide chip and real-time PCR detected mixed genotypes B and C in 9 samples. Realtime PCR was the rapidest and cheapest among the three assays.CONCLUSION: Oligonucleotide chip and real-time PCR are able to detect mixed genotypes, while sequencing only detects the dominant genotype in clinical samples.

  11. Improving signal intensities for genes with low-expression on oligonucleotide microarrays

    Directory of Open Access Journals (Sweden)

    Hu Limei


    Full Text Available Abstract Background DNA microarrays using long oligonucleotide probes are widely used to evaluate gene expression in biological samples. These oligonucleotides are pre-synthesized and sequence-optimized to represent specific genes with minimal cross-hybridization to homologous genes. Probe length and concentration are critical factors for signal sensitivity, particularly when genes with various expression levels are being tested. We evaluated the effects of oligonucleotide probe length and concentration on signal intensity measurements of the expression levels of genes in a target sample. Results Selected genes of various expression levels in a single cell line were hybridized to oligonucleotide arrays of four lengths and four concentrations of probes to determine how these critical parameters affected the intensity of the signal representing their expression. We found that oligonucleotides of longer length significantly increased the signals of genes with low-expression in the target. High-expressing gene signals were also boosted but to a lesser degree. Increasing the probe concentration, however, did not linearly increase the signal intensity for either low- or high-expressing genes. Conclusions We conclude that the longer the oligonuclotide probe the better the signal intensities of low expressing genes on oligonucleotide arrays.

  12. Porous silicon-cell penetrating peptide hybrid nanocarrier for intracellular delivery of oligonucleotides. (United States)

    Rytkönen, Jussi; Arukuusk, Piret; Xu, Wujun; Kurrikoff, Kaido; Langel, Ulo; Lehto, Vesa-Pekka; Närvänen, Ale


    The largest obstacle to the use of oligonucleotides as therapeutic agents is the delivery of these large and negatively charged biomolecules through cell membranes into intracellular space. Mesoporous silicon (PSi) is widely recognized as a potential material for drug delivery purposes due to its several beneficial features like large surface area and pore volume, high loading capacity, biocompatibility, and biodegradability. In the present study, PSi nanoparticles stabilized by thermal oxidation or thermal carbonization and subsequently modified by grafting aminosilanes on the surface are utilized as an oligonucleotide carrier. Splice correcting oligonucleotides (SCOs), a model oligonucleotide drug, were loaded into the positively charged PSi nanoparticles with a loading degree as high as 14.3% (w/w). Rapid loading was achieved by electrostatic interactions, with the loading efficiencies reaching 100% within 5 min. The nanoparticles were shown to deliver and release SCOs, in its biologically active form, inside cells when formulated together with cell penetrating peptides (CPP). The biological effect was monitored with splice correction assay and confocal microscopy utilizing HeLa pLuc 705 cells. Furthermore, the use of PSi carrier platform in oligonucleotide delivery did not reduce the cell viability. Additionally, the SCO-CPP complexes formed in the pores of the carrier were stabilized against proteolytic digestion. The advantageous properties of protecting and releasing the cargo and the possibility to further functionalize the carrier surface make the hybrid nanoparticles a potential system for oligonucleotide delivery.

  13. Basic Research Firing Facility (United States)

    Federal Laboratory Consortium — The Basic Research Firing Facility is an indoor ballistic test facility that has recently transitioned from a customer-based facility to a dedicated basic research...

  14. DNA sequence analysis by hybridization with oligonucleotide microchips : MALDI mass spectrometry identification of 5mers contiguously stacked to microchip oligonucleotides.

    Energy Technology Data Exchange (ETDEWEB)

    Stomakhin, A. A.; Vasiliskov, V. A.; Timofeev, E.; Schulga, D.; Cotter, R. J.; Mirzabekov, A. D.; Biochip Technology Center; Engelhardt Inst. of Molecular Biology; Moscow Inst. of Physics and Technology; Middle Atlantic Mass Spectrometry Lab.; Johns Hopkins Univ. School of Medicine


    Matrix-assisted laser desorption ionization mass spectrometry (MALDI MS) has been applied to increase the informational output from DNA sequence analysis. It has been used to analyze DNA by hybridization with microarrays of gel-immobilized oligonucleotides extended with stacked 5mers. In model experiments, a 28 nt long DNA fragment was hybridized with 10 immobilized, overlapping 8mers. Then, in a second round of hybridization DNA-8mer duplexes were hybridized with a mixture of 10 5mers. The stability of the 5mer complex with DNA was increased to raise the melting temperature of the duplex by 10-15{sup o}C as a result of stacking interaction with 8mers. Contiguous 13 bp duplexes containing an internal break were formed. MALDI MS identified one or, in some cases, two 5mers contiguously stacked to each DNA-8mer duplex formed on the microchip. Incorporating a mass label into 5mers optimized MALDI MS monitoring. This procedure enabled us to reconstitute the sequence of a model DNA fragment and identify polymorphic nucleotides. The application of MALDI MS identification of contiguously stacked 5mers to increase the length of DNA for sequence analysis is discussed.

  15. Body Basics Library (United States)

    ... of Healthy Breakfasts Shyness About the Body Basics Library KidsHealth > For Teens > About the Body Basics Library A A A Did you ever wonder what ... system, part, and process works. Use this medical library to find out about basic human anatomy, how ...

  16. Body Basics Library (United States)

    ... of Healthy Breakfasts Shyness About the Body Basics Library KidsHealth > For Teens > About the Body Basics Library Print A A A Did you ever wonder ... system, part, and process works. Use this medical library to find out about basic human anatomy, how ...

  17. Basic Cake Decorating Workbook. (United States)

    Bogdany, Mel

    Included in this student workbook for basic cake decorating are the following: (1) Drawings of steps in a basic way to ice a layer cake, how to make a paper cone, various sizes of flower nails, various sizes and types of tin pastry tubes, and special rose tubes; (2) recipes for basic decorating icings (buttercream, rose paste, and royal icing);…

  18. 75 FR 78725 - Recreational Boating Safety Projects, Programs and Activities Funded Under Provisions of the... (United States)


    ... Guard from the Sport Fish Restoration and Boating Trust Fund and committed, obligated, and/or expended... defect. ($17,335). Web-based Document Management System: Funding was provided to continue to provide a web-based document management system to better enable the handling of thousands of...

  19. 'We get nothing from fishing' : Fishing for boat opportunities amongst Senegalese fisher migrants

    NARCIS (Netherlands)

    Nyamnjoh, Henrietta Mambo


    The world is regularly confronted in the media with dramatic images of African boat migrants. Seemingly desperate, these Africans, most of them males, are willing to risk a perilous journey at sea, hoping for a better life in Europe. And, even worse, hundreds more are believed to die each year, swal

  20. BOAT: automatic alignment of biomedical ontologies using term informativeness and candidate selection. (United States)

    Chua, Watson Wei Khong; Kim, Jung-Jae


    The biomedical sciences is one of the few domains where ontologies are widely being developed to facilitate information retrieval and knowledge sharing, but there still remains the problem that applications using different ontologies cannot share knowledge without explicit references between overlapping concepts. Ontology alignment is the task of identifying such equivalence relations between concepts across ontologies. Its application to the biomedical domain should address two open issues: (1) determining the equivalence of concept-pairs which have overlapping terms in their names, and (2) the high run-time required to align large ontologies which are typical in the biomedical domain. To address them, we present a novel approach, named the Biomedical Ontologies Alignment Technique (BOAT), which is state-of-the-art in terms of F-measure, precision and speed. A key feature of BOAT is that it considers the informativeness of each component word in the concept labels, which has significant impact on biomedical ontologies, resulting in a 12.2% increase in F-measure. Another important feature of BOAT is that it selects for comparison only concept pairs that show high likelihoods of equivalence, based on the similarity of their annotations. BOAT's F-measure of 0.88 for the alignment of the mouse and human anatomy ontologies is on par with that of another state-of-the-art matcher, AgreementMaker, while taking a shorter time.

  1. Motion induced passenger dislocations in life raft and life boat configurations

    NARCIS (Netherlands)

    Bos, J.E.; Andersen ,O.; Reinke, P.


    Within the European project SafeCrafts, dealing with the safe abandoning of ships, the dislocation of passengers aboard life boats and rafts has been considered to be a risk regarding passenger survival at sea. To get an idea about the risks and the affecting factors involved, we studied the effects

  2. Simple Verification of the Parabolic Shape of a Rotating Liquid and a Boat on Its Surface (United States)

    Sabatka, Z.; Dvorak, L.


    This article describes a simple and inexpensive way to create and to verify the parabolic surface of a rotating liquid. The liquid is water. The second part of the article deals with the problem of a boat on the surface of a rotating liquid. (Contains 1 table, 10 figures and 5 footnotes.)

  3. 75 FR 43823 - Safety Zone; He'eia Kea Small Boat Harbor, Kaneohe Bay, Oahu, HI (United States)


    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF HOMELAND SECURITY Coast Guard 33 CFR Part 165 RIN 1625-AA00 Safety Zone; He'eia Kea Small Boat Harbor, Kaneohe Bay, Oahu, HI AGENCY: Coast Guard, DHS. ACTION: Temporary Final Rule. SUMMARY: The Coast Guard...

  4. 33 CFR 100.111 - Stonington Lobster Boat Races, Stonington, ME. (United States)


    ... 33 Navigation and Navigable Waters 1 2010-07-01 2010-07-01 false Stonington Lobster Boat Races, Stonington, ME. 100.111 Section 100.111 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF HOMELAND SECURITY REGATTAS AND MARINE PARADES SAFETY OF LIFE ON NAVIGABLE WATERS § 100.111 Stonington Lobster...

  5. 33 CFR 100.110 - World's Fastest Lobster Boat Race, Jonesport, ME. (United States)


    ... 33 Navigation and Navigable Waters 1 2010-07-01 2010-07-01 false World's Fastest Lobster Boat Race, Jonesport, ME. 100.110 Section 100.110 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF HOMELAND SECURITY REGATTAS AND MARINE PARADES SAFETY OF LIFE ON NAVIGABLE WATERS § 100.110 World's Fastest...

  6. 46 CFR 199.175 - Survival craft and rescue boat equipment. (United States)


    ....75 inches) below the boat's light waterline. (19) Mirror. The signalling mirror must be approved... Knife 1,4 1 1 1 1 18 Ladder 1 1 1 1 19 Mirror, signalling 1 1 1 1 20 Oars, units 5,6 1 1 1 1 Paddles 2...

  7. The use of vascular plants as traditional boat raw material by Yachai tribe in Mappi Regency

    Directory of Open Access Journals (Sweden)



    Full Text Available This research is executed aim to know the plant species and the way of exploiting permanent wood upon which traditional boat making by Yachai tribe in Mappi regency. The Method that used in this research is descriptive method with the structural semi interview technique and direct perception in field. Result of research indicate that the tribe Yachai exploit the plant species have permanent wood upon which traditional boat as much 26 species from 14 family. There are 8 wood species which is often used for the body of boat and also own the good quality according to Yachai tribe, that is Atam (Scihizomeria serrata Hochr, Batki (Adinandra forbesii Baker. F, Chomach (Gordonia papuana Kobuski, Rupke (Tristania sp., Bao (Dillenia papuana artelli, Top (Buchanania macrocarpa Laut, Mitbo (Cordia Dichtoma Forst., and Yunun (Camnosperma brevipetiolata Volkens. While to part of oar exploit 2 wood species that is Bach (Buchanania Arborescens.Bi and Tup (Litsea ampala Merr. Yachai Tribe recognized 3 boat model owning different size measure and function, that is Junun Ramchai, Junun Pochoi and Junun Toch.

  8. Stone sculptures of goddesses on the boats from Goa, west coast of India

    Digital Repository Service at National Institute of Oceanography (India)

    Gaur, A; Kerkar, R.

    goddess panels. A few sculptures reflect influence of folk art traditions in Goa. On the basis of lion figures in a few sculptured panels indicate that these boat motifs must have been engraved during the Kadamba period (10 th - 13 th) when Goa witnessed...

  9. The boat people of Vietnam. Rescue on the South China Sea. (United States)

    Hayes, T J; Thomas, D; Dewar, E P


    Poor economic conditions in Vietnam have encouraged large numbers of people seeking a better life in the West to join the flow of political refugees leaving the country as "boat people". We report the events surrounding one such group of people who were rescued from the South China Sea by the Outback 88 Task Group.

  10. Correction of a Cystic Fibrosis Splicing Mutation by Antisense Oligonucleotides. (United States)

    Igreja, Susana; Clarke, Luka A; Botelho, Hugo M; Marques, Luís; Amaral, Margarida D


    Cystic fibrosis (CF), the most common life-threatening genetic disease in Caucasians, is caused by ∼2,000 different mutations in the CF transmembrane conductance regulator (CFTR) gene. A significant fraction of these (∼13%) affect pre-mRNA splicing for which novel therapies have been somewhat neglected. We have previously described the effect of the CFTR splicing mutation c.2657+5G>A in IVS16, showing that it originates transcripts lacking exon 16 as well as wild-type transcripts. Here, we tested an RNA-based antisense oligonucleotide (AON) strategy to correct the aberrant splicing caused by this mutation. Two AONs (AON1/2) complementary to the pre-mRNA IVS16 mutant region were designed and their effect on splicing was assessed at the RNA and protein levels, on intracellular protein localization and function. To this end, we used the 2657+5G>A mutant CFTR minigene stably expressed in HEK293 Flp-In cells that express a single copy of the transgene. RNA data from AON1-treated mutant cells show that exon 16 inclusion was almost completely restored (to 95%), also resulting in increased levels of correctly localized CFTR protein at the plasma membrane (PM) and with increased function. A novel two-color CFTR splicing reporter minigene developed here allowed the quantitative monitoring of splicing by automated microscopy localization of CFTR at the PM. The AON strategy is thus a promising therapeutic approach for the specific correction of alternative splicing.

  11. Profiled support vector machines for antisense oligonucleotide efficacy prediction

    Directory of Open Access Journals (Sweden)

    Martín-Guerrero José D


    Full Text Available Abstract Background This paper presents the use of Support Vector Machines (SVMs for prediction and analysis of antisense oligonucleotide (AO efficacy. The collected database comprises 315 AO molecules including 68 features each, inducing a problem well-suited to SVMs. The task of feature selection is crucial given the presence of noisy or redundant features, and the well-known problem of the curse of dimensionality. We propose a two-stage strategy to develop an optimal model: (1 feature selection using correlation analysis, mutual information, and SVM-based recursive feature elimination (SVM-RFE, and (2 AO prediction using standard and profiled SVM formulations. A profiled SVM gives different weights to different parts of the training data to focus the training on the most important regions. Results In the first stage, the SVM-RFE technique was most efficient and robust in the presence of low number of samples and high input space dimension. This method yielded an optimal subset of 14 representative features, which were all related to energy and sequence motifs. The second stage evaluated the performance of the predictors (overall correlation coefficient between observed and predicted efficacy, r; mean error, ME; and root-mean-square-error, RMSE using 8-fold and minus-one-RNA cross-validation methods. The profiled SVM produced the best results (r = 0.44, ME = 0.022, and RMSE= 0.278 and predicted high (>75% inhibition of gene expression and low efficacy ( Conclusions The SVM approach is well suited to the AO prediction problem, and yields a prediction accuracy superior to previous methods. The profiled SVM was found to perform better than the standard SVM, suggesting that it could lead to improvements in other prediction problems as well.

  12. Application of heteronuclear couplings to conformational analysis of oligonucleotides

    Energy Technology Data Exchange (ETDEWEB)

    Zhu, G. [Univ. of Maryland, College Park, MD (United States); Live, D. [Memorial Sloan-Kettering Cancer Center, New York, NY (United States); Bax, A. [NIDDK National Institutes of Health, Bethesda, MD (United States)


    The value of vicinal coupling constants extracted from NMR spectra in deducing torsion angles for conformational analysis is well recognized. Due to the abundance of protons, their couplings have been mostly widely used. In many instances, couplings between protons and other nuclei may be a valuable complement to proton-proton couplings or, in some instances, may be the only coupling available to characterize the torsion angle about a bond. Recently, heteronuclear couplings have been used to great benefit in studies of isotopically enriched proteins, and this general approach has been extended to peptides at natural abundance. The possibility of using this approach to study oligonucleotides is also attractive but has not as yet been widely exploited. With the development of strategies for labeling such molecules, particularly RNAs, this may become an important component in conformational analysis. For DNA, labeling is less accessible, but sufficient quantities of unlabeled material are readily available for measuring these couplings at natural abundance. We chose several DNA systems to explore the usefulness of heteronuclear couplings in addressing the sugar conformation and the glycosidic torsion angle. Intensities of cross peaks in long-range HMQC experiments can be related to the couplings. Crosspeaks involving H1{prime} and C1{prime} atoms have been emphasized because of the superior shift dispersion at these positions between sugar protons and carbon atoms. Results will be shown for the self-complementary Dickerson duplex dodecamer sequence d(CGCGAATTCGCG) and for d(GGTCGG), which dimerizes to form a G-tetrad structure incorporating both syn and anti base orientations. The couplings provide a clear discrimination between presence of C3{prime}-endo and C2{prime}-endo conformations of the sugars and syn and anti bases arrangements.

  13. Evaluation of ADCP apparent bed load velocity in a large sand-bed river: Moving versus stationary boat conditions (United States)

    Jamieson, E.C.; Rennie, C.D.; Jacobson, R.B.; Townsend, R.D.


    Detailed mapping of bathymetry and apparent bed load velocity using a boat-mounted acoustic Doppler current profiler (ADCP) was carried out along a 388-m section of the lower Missouri River near Columbia, Missouri. Sampling transects (moving boat) were completed at 5- and 20-m spacing along the study section. Stationary (fixed-boat) measurements were made by maintaining constant boat position over a target point where the position of the boat did not deviate more than 3 m in any direction. For each transect and stationary measurement, apparent bed load velocity (vb) was estimated using ADCP bottom tracking data and high precision real-time kinematic (RTK) global positioning system (GPS). The principal objectives of this research are to (1) determine whether boat motion introduces a bias in apparent bed load velocity measurements; and (2) evaluate the reliability of ADCP bed velocity measurements for a range of sediment transport environments. Results indicate that both high transport (vb>0.6 m/s) and moving-boat conditions (for both high and low transport environments) increase the relative variability in estimates of mean bed velocity. Despite this, the spatially dense single-transect measurements were capable of producing detailed bed velocity maps that correspond closely with the expected pattern of sediment transport over large dunes. ?? 2011 American Society of Civil Engineers.

  14. A simple and rapid method for the preparation of homologous DNA oligonucleotide hybridization probes from heterologous gene sequences and probes. (United States)

    Maxwell, E S; Sarge, K D


    We describe a simple and rapid method for the preparation of homologous DNA oligonucleotide probes for hybridization analysis and/or cDNA/genomic library screening. With this method, a synthetic DNA oligonucleotide derived from a known heterologous DNA/RNA/protein sequence is annealed to an RNA preparation containing the gene transcript of interest. Any unpaired 3'-terminal oligonucleotides of the heterologous DNA primer are then removed using the 3' exonuclease activity of the DNA Polymerase I Klenow fragment before primer extension/dideoxynucleotide sequencing of the annealed RNA species with AMV reverse transcriptase. From the determined RNA sequence, a completely homologous DNA oligonucleotide probe is then prepared. This approach has been used to prepare a homologous DNA oligonucleotide probe for the successful library screening of the yeast hybRNA gene starting with a heterologous mouse hybRNA DNA oligonucleotide probe.

  15. 'Specific' oligonucleotides often recognize more than one gene: the limits of in situ hybridization applied to GABA receptors. (United States)

    Mladinic, M; Didelon, F; Cherubini, E; Bradbury, A


    As exquisite probes for gene sequences, oligonucleotides are one of the most powerful tools of recombinant molecular biology. In studying the GABA receptor subunits in the neonatal hippocampus we have used oligonucleotide probes in in situ hybridization and cloning techniques. The oligonucleotides used and assumed to be specific for the target gene, actually recognized more than one gene, leading to surprising and contradictory results. In particular, we found that a GABA(A)-rho specific oligonucleotide recognized an abundant, previously unknown, transcription factor in both in situ and library screening, while oligos 'specific' for GABA(A) subunits were able to recognize 30 additional unrelated genes in library screening. This suggests that positive results obtained with oligonucleotides should be interpreted with caution unless confirmed by identical results with oligonucleotides from different parts of the same gene, or cDNA library screening excludes the presence of other hybridizing species.

  16. Diffusion of Oligonucleotides from within Iron-Crosslinked Polyelectrolyte-Modified Alginate Beads: A Model System for Drug Release

    CERN Document Server

    Privman, Vladimir; Luz, Roberto A S; Guz, Nataliia; Glasser, M Lawrence; Katz, Evgeny


    We developed and experimentally verified an analytical model to describe diffusion of oligonucleotides from stable hydrogel beads. The synthesized alginate beads are Fe3+-cross-linked as well as polyelectrolyte-doped for uniformity and stability at physiological pH. Data on diffusion of oligonucleotides from inside the beads provide physical insights into the volume nature of the immobilization of a fraction of oligonucleotides due to polyelectrolyte cross-linking, i.e., the absence of the surface-layer barrier in this case. Furthermore, our results suggest a new simple approach to measuring the diffusion coefficient of the mobile oligonucleotide molecules inside hydrogel. The considered alginate beads provide a model for a well-defined component in drug release systems and for the oligonucleotide-release transduction steps in drug-delivering and biocomputing applications. This is illustrated by destabilizing the beads with citrate that induces full oligonucleotide release with non-diffusional kinetics.

  17. Analyses of point mutation repair and allelic heterogeneity generated by CRISPR/Cas9 and single-stranded DNA oligonucleotides


    Pawel Bialk; Brett Sansbury; Natalia Rivera-Torres; Kevin Bloh; Dula Man; Kmiec, Eric B.


    The repair of a point mutation can be facilitated by combined activity of a single-stranded oligonucleotide and a CRISPR/Cas9 system. While the mechanism of action of combinatorial gene editing remains to be elucidated, the regulatory circuitry of nucleotide exchange executed by oligonucleotides alone has been largely defined. The presence of the appropriate CRISPR/Cas9 system leads to an enhancement in the frequency of gene editing directed by single-stranded DNA oligonucleotides. While CRIS...

  18. Basics of Bayesian Learning - Basically Bayes

    DEFF Research Database (Denmark)

    Larsen, Jan

    Tutorial presented at the IEEE Machine Learning for Signal Processing Workshop 2006, Maynooth, Ireland, September 8, 2006. The tutorial focuses on the basic elements of Bayesian learning and its relation to classical learning paradigms. This includes a critical discussion of the pros and cons...

  19. Experimental analysis of oligonucleotide microarray design criteria to detect deletions by comparative genomic hybridization

    Directory of Open Access Journals (Sweden)

    Moerman Donald G


    Full Text Available Abstract Background Microarray comparative genomic hybridization (CGH is currently one of the most powerful techniques to measure DNA copy number in large genomes. In humans, microarray CGH is widely used to assess copy number variants in healthy individuals and copy number aberrations associated with various diseases, syndromes and disease susceptibility. In model organisms such as Caenorhabditis elegans (C. elegans the technique has been applied to detect mutations, primarily deletions, in strains of interest. Although various constraints on oligonucleotide properties have been suggested to minimize non-specific hybridization and improve the data quality, there have been few experimental validations for CGH experiments. For genomic regions where strict design filters would limit the coverage it would also be useful to quantify the expected loss in data quality associated with relaxed design criteria. Results We have quantified the effects of filtering various oligonucleotide properties by measuring the resolving power for detecting deletions in the human and C. elegans genomes using NimbleGen microarrays. Approximately twice as many oligonucleotides are typically required to be affected by a deletion in human DNA samples in order to achieve the same statistical confidence as one would observe for a deletion in C. elegans. Surprisingly, the ability to detect deletions strongly depends on the oligonucleotide 15-mer count, which is defined as the sum of the genomic frequency of all the constituent 15-mers within the oligonucleotide. A similarity level above 80% to non-target sequences over the length of the probe produces significant cross-hybridization. We recommend the use of a fairly large melting temperature window of up to 10°C, the elimination of repeat sequences, the elimination of homopolymers longer than 5 nucleotides, and a threshold of -1 kcal/mol on the oligonucleotide self-folding energy. We observed very little difference in data

  20. Basic molecular spectroscopy

    CERN Document Server

    Gorry, PA


    BASIC Molecular Spectroscopy discusses the utilization of the Beginner's All-purpose Symbolic Instruction Code (BASIC) programming language in molecular spectroscopy. The book is comprised of five chapters that provide an introduction to molecular spectroscopy through programs written in BASIC. The coverage of the text includes rotational spectra, vibrational spectra, and Raman and electronic spectra. The book will be of great use to students who are currently taking a course in molecular spectroscopy.

  1. Basic digital signal processing

    CERN Document Server

    Lockhart, Gordon B


    Basic Digital Signal Processing describes the principles of digital signal processing and experiments with BASIC programs involving the fast Fourier theorem (FFT). The book reviews the fundamentals of the BASIC program, continuous and discrete time signals including analog signals, Fourier analysis, discrete Fourier transform, signal energy, power. The text also explains digital signal processing involving digital filters, linear time-variant systems, discrete time unit impulse, discrete-time convolution, and the alternative structure for second order infinite impulse response (IIR) sections.

  2. Efficient gene silencing by delivery of locked nucleic acid antisense oligonucleotides, unassisted by transfection reagents. (United States)

    Stein, C A; Hansen, J Bo; Lai, Johnathan; Wu, SiJian; Voskresenskiy, Anatoliy; Høg, Anja; Worm, Jesper; Hedtjärn, Maj; Souleimanian, Naira; Miller, Paul; Soifer, Harris S; Castanotto, Daniella; Benimetskaya, Luba; Ørum, Henrik; Koch, Troels


    For the past 15-20 years, the intracellular delivery and silencing activity of oligodeoxynucleotides have been essentially completely dependent on the use of a delivery technology (e.g. lipofection). We have developed a method (called 'gymnosis') that does not require the use of any transfection reagent or any additives to serum whatsoever, but rather takes advantage of the normal growth properties of cells in tissue culture in order to promote productive oligonucleotide uptake. This robust method permits the sequence-specific silencing of multiple targets in a large number of cell types in tissue culture, both at the protein and mRNA level, at concentrations in the low micromolar range. Optimum results were obtained with locked nucleic acid (LNA) phosphorothioate gap-mers. By appropriate manipulation of oligonucleotide dosing, this silencing can be continuously maintained with little or no toxicity for >240 days. High levels of oligonucleotide in the cell nucleus are not a requirement for gene silencing, contrary to long accepted dogma. In addition, gymnotic delivery can efficiently deliver oligonucleotides to suspension cells that are known to be very difficult to transfect. Finally, the pattern of gene silencing of in vitro gymnotically delivered oligonucleotides correlates particularly well with in vivo silencing. The establishment of this link is of particular significance to those in the academic research and drug discovery and development communities.

  3. Novel Efficient Cell-Penetrating, Peptide-Mediated Strategy for Enhancing Telomerase Inhibitor Oligonucleotides. (United States)

    Muñoz-Alarcón, Andrés; Eriksson, Jonas; Langel, Ülo


    At present, there are several therapeutic approaches for targeting telomerase in tumors. One in particular, currently undergoing clinical trials, is based on synthetic lipid-modified oligonucleotide antagonists aimed at inhibiting the ribonucleoprotein subunit of human telomerase. However, while enabling efficient uptake, the lipid modifications reduce the potency of the therapeutic oligonucleotides compared to nonmodified oligonucleotides. Moreover, lipid modification may increase oligonucleotide accumulation in the liver causing undesirable hepatotoxicity. Noncovalent complexation strategies for cell-penetrating peptide (CPP)-mediated delivery present an option to circumvent the need for potency-reducing modifications, while allowing for a highly efficient uptake, and could significantly improve the efficiency of telomerase-targeting cancer therapeutics. Delivery of a nonlipidated locked nucleic acid/2'-O-methyl mixmer significantly inhibits the telomerase activity in treated HeLa cells. The inhibitory effect was further improved through addition of a CPP. Furthermore, calculated IC50-values for the oligonucleotide delivered by CPPs into HeLa cells are more than 20 times lower than telomerase inhibitor Imetelstat, currently undergoing clinical trials. These results emphasize the potential of CPP-mediated delivery of future pharmaceuticals and provide means by which to enhance an already promising therapeutic strategy for cancer treatment.

  4. Conjugates of Phthalocyanines With Oligonucleotides as Reagents for Sensitized or Catalytic DNA Modification

    Directory of Open Access Journals (Sweden)


    Full Text Available Several conjugates of metallophthalocyanines with deoxyribooligonucleotides were synthesized to investigate sequence-specific modification of DNA by them. Oligonucleotide parts of these conjugates were responsible for the recognition of selected complementary sequences on the DNA target. Metallophthalocyanines were able to induce the DNA modification: phthalocyanines of Zn(II and Al(III were active as photosensitizers in the generation of singlet oxygen 1 O 2 , while phthalocyanine of Co(II promoted DNA oxidation by molecular oxygen through the catalysis of formation of reactive oxygen species ( ⋅ O 2 − , O 2 H 2 , OH. Irradiation of the reaction mixture containing either Zn(II- or Al(III-tetracarboxyphthalocyanine conjugates of oligonucleotide pd(TCTTCCCA with light of > 340 nm wavelength (Hg lamp or He/Ne laser resulted in the modification of the 22-nucleotide target d(TGAATGGGAAGAGGGTCAGGTT. A conjugate of Co(II-tetracarboxyphthalocyanine with the oligonucleotide was found to modify the DNA target in the presence of O 2 and 2-mercaptoethanol or in the presence of O 2 H 2 . Under both sensitized and catalyzed conditions, the nucleotides G 13 – G 15 were mainly modified, providing evidence that the reaction proceeded in the double-stranded oligonucleotide. These results suggest the possible use of phthalocyanine-oligonucleotide conjugates as novel artificial regulators of gene expression and therapeutic agents for treatment of cancer.

  5. Oligonucleotide-induced alternative splicing of serotonin 2C receptor reduces food intake. (United States)

    Zhang, Zhaiyi; Shen, Manli; Gresch, Paul J; Ghamari-Langroudi, Masoud; Rabchevsky, Alexander G; Emeson, Ronald B; Stamm, Stefan


    The serotonin 2C receptor regulates food uptake, and its activity is regulated by alternative pre-mRNA splicing. Alternative exon skipping is predicted to generate a truncated receptor protein isoform, whose existence was confirmed with a new antiserum. The truncated receptor sequesters the full-length receptor in intracellular membranes. We developed an oligonucleotide that promotes exon inclusion, which increases the ratio of the full-length to truncated receptor protein. Decreasing the amount of truncated receptor results in the accumulation of full-length, constitutively active receptor at the cell surface. After injection into the third ventricle of mice, the oligonucleotide accumulates in the arcuate nucleus, where it changes alternative splicing of the serotonin 2C receptor and increases pro-opiomelanocortin expression. Oligonucleotide injection reduced food intake in both wild-type and ob/ob mice. Unexpectedly, the oligonucleotide crossed the blood-brain barrier and its systemic delivery reduced food intake in wild-type mice. The physiological effect of the oligonucleotide suggests that a truncated splice variant regulates the activity of the serotonin 2C receptor, indicating that therapies aimed to change pre-mRNA processing could be useful to treat hyperphagia, characteristic for disorders like Prader-Willi syndrome.

  6. A facile inhibitor screening of SARS coronavirus N protein using nanoparticle-based RNA oligonucleotide

    Directory of Open Access Journals (Sweden)

    Roh C


    Full Text Available Changhyun RohDivision of Biotechnology, Advanced Radiation Technology Institute (ARTI, Korea Atomic Energy Research Institute (KAERI, Jeongeup, Republic of KoreaAbstract: Hundreds of million people worldwide have been infected with severe acute respiratory syndrome (SARS, and the rate of global death from SARS has remarkably increased. Hence, the development of efficient drug treatments for the biological effects of SARS is highly needed. We have previously shown that quantum dots (QDs-conjugated RNA oligonucleotide is sensitive to the specific recognition of the SARS-associated coronavirus (SARS-CoV nucleocapsid (N protein. In this study, we found that a designed biochip could analyze inhibitors of the SARS-CoV N protein using nanoparticle-based RNA oligonucleotide. Among the polyphenolic compounds examined, (--catechin gallate and (--gallocatechin gallate demonstrated a remarkable inhibition activity on SARS-CoV N protein. (--catechin gallate and (--gallocatechin gallate attenuated the binding affinity in a concentrated manner as evidenced by QDs-conjugated RNA oligonucleotide on a designed biochip. At a concentration of 0.05 µg mL–1, (--catechin gallate and (--gallocatechin gallate showed more than 40% inhibition activity on a nanoparticle-based RNA oligonucleotide biochip system.Keywords: SARS, RNA oligonucleotide, quantum dots, inhibitor, screening

  7. Aspects of oligonucleotide and peptide sequencing with MALDI and electrospray mass spectrometry. (United States)

    Owens, D R; Bothner, B; Phung, Q; Harris, K; Siuzdak, G


    Biopolymer sequencing with mass spectrometry has become increasingly important and accessible with the development of matrix-assisted laser desorption/ionization (MALDI) and electrospray ionization (ESI). Here we examine the use of sequential digestion for the rapid identification of proteolytic fragments, in turn highlighting the general utility of enzymatic MALDI ladder sequencing and ESI tandem mass spectrometry. Analyses were performed on oligonucleotides ranging in size from 2 to 50 residues, on peptides ranging in size from 7 to 44 residues and on viral coat proteins. MALDI ladder sequencing using exonuclease digestion generated a uniform distribution of ions and provided complete sequence information on the oligonucleotides 2-30 nucleic acid residues long. Only partial sequence information was obtained on the longer oligonucleotides. C-terminal peptide ladder sequencing typically provided information from 4 to 7 amino acids into the peptide. Sequential digestion, or endoprotease followed by exoprotease exposure, was also successfully applied to a trypsin digest of viral proteins. Analysis of ladder sequenced peptides by LCMS generated less information than in the MALDI-MS analysis and ESI-MS2 normally provided partial sequence information on both the small oligonucleotides and peptides. In general, MALDI ladder sequencing offered information on a broader mass range of biopolymers than ESI-MS2 and was relatively straightforward to interpret, especially for oligonucleotides.

  8. LNA-modified isothermal oligonucleotide microarray for differentiating bacilli of similar origin

    Indian Academy of Sciences (India)

    Jing Yan; Ying Yuan; Runqing Mu; Hong Shang; Yifu Guan


    Oligonucleotide microarray has been one of the most powerful tools in the ‘Post-Genome Era’ for its high sensitivity, high throughput and parallel processing capability. To achieve high detection specificity, we fabricated an isothermal microarray using locked nucleic acid (LNA)-modified oligonucleotide probes, since LNA has demonstrated the advanced ability to enhance the binding affinity toward their complementary nucleotides. After designing the nucleotide sequences of these oligonucleotide probes for gram-positive bacilli of similar origin (Bacillus subtilis, Bacillus licheniformis, Bacillus pumilus, Bacillus megaterium and Bacillus circulans), we unified the melting temperatures of these oligonucleotide probes by modifying some nucleotides using LNA. Furthermore, we optimized the experimental procedures of hydrating microarray slides, blocking side surface as well as labelling the PCR products. Experimental results revealed that KOD Dash DNA polymerase could efficiently incorporate Cy3-dCTP into the PCR products, and the LNA-isothermal oligonucleotide microarray were able to distinguish the bacilli of similar origin with a high degree of accuracy and specificity under the optimized experimental condition.

  9. Combined in vitro transcription and reverse transcription to amplify and label complex synthetic oligonucleotide probe libraries. (United States)

    Murgha, Yusuf; Beliveau, Brian; Semrau, Kassandra; Schwartz, Donald; Wu, Chao-Ting; Gulari, Erdogan; Rouillard, Jean-Marie


    Oligonucleotide microarrays allow the production of complex custom oligonucleotide libraries for nucleic acid detection-based applications such as fluorescence in situ hybridization (FISH). We have developed a PCR-free method to make single-stranded DNA (ssDNA) fluorescent probes through an intermediate RNA library. A double-stranded oligonucleotide library is amplified by transcription to create an RNA library. Next, dye- or hapten-conjugate primers are used to reverse transcribe the RNA to produce a dye-labeled cDNA library. Finally the RNA is hydrolyzed under alkaline conditions to obtain the single-stranded fluorescent probes library. Starting from unique oligonucleotide library constructs, we present two methods to produce single-stranded probe libraries. The two methods differ in the type of reverse transcription (RT) primer, the incorporation of fluorescent dye, and the purification of fluorescent probes. The first method employs dye-labeled reverse transcription primers to produce multiple differentially single-labeled probe subsets from one microarray library. The fluorescent probes are purified from excess primers by oligonucleotide-bead capture. The second method uses an RNA:DNA chimeric primer and amino-modified nucleotides to produce amino-allyl probes. The excess primers and RNA are hydrolyzed under alkaline conditions, followed by probe purification and labeling with amino-reactive dyes. The fluorescent probes created by the combination of transcription and reverse transcription can be used for FISH and to detect any RNA and DNA targets via hybridization.

  10. Efficient oligonucleotide probe selection for pan-genomic tiling arrays

    Directory of Open Access Journals (Sweden)

    Zhang Wei


    Full Text Available Abstract Background Array comparative genomic hybridization is a fast and cost-effective method for detecting, genotyping, and comparing the genomic sequence of unknown bacterial isolates. This method, as with all microarray applications, requires adequate coverage of probes targeting the regions of interest. An unbiased tiling of probes across the entire length of the genome is the most flexible design approach. However, such a whole-genome tiling requires that the genome sequence is known in advance. For the accurate analysis of uncharacterized bacteria, an array must query a fully representative set of sequences from the species' pan-genome. Prior microarrays have included only a single strain per array or the conserved sequences of gene families. These arrays omit potentially important genes and sequence variants from the pan-genome. Results This paper presents a new probe selection algorithm (PanArray that can tile multiple whole genomes using a minimal number of probes. Unlike arrays built on clustered gene families, PanArray uses an unbiased, probe-centric approach that does not rely on annotations, gene clustering, or multi-alignments. Instead, probes are evenly tiled across all sequences of the pan-genome at a consistent level of coverage. To minimize the required number of probes, probes conserved across multiple strains in the pan-genome are selected first, and additional probes are used only where necessary to span polymorphic regions of the genome. The viability of the algorithm is demonstrated by array designs for seven different bacterial pan-genomes and, in particular, the design of a 385,000 probe array that fully tiles the genomes of 20 different Listeria monocytogenes strains with overlapping probes at greater than twofold coverage. Conclusion PanArray is an oligonucleotide probe selection algorithm for tiling multiple genome sequences using a minimal number of probes. It is capable of fully tiling all genomes of a species on

  11. A novel multifunctional oligonucleotide microarray for Toxoplasma gondii

    Directory of Open Access Journals (Sweden)

    Chen Feng


    Full Text Available Abstract Background Microarrays are invaluable tools for genome interrogation, SNP detection, and expression analysis, among other applications. Such broad capabilities would be of value to many pathogen research communities, although the development and use of genome-scale microarrays is often a costly undertaking. Therefore, effective methods for reducing unnecessary probes while maintaining or expanding functionality would be relevant to many investigators. Results Taking advantage of available genome sequences and annotation for Toxoplasma gondii (a pathogenic parasite responsible for illness in immunocompromised individuals and Plasmodium falciparum (a related parasite responsible for severe human malaria, we designed a single oligonucleotide microarray capable of supporting a wide range of applications at relatively low cost, including genome-wide expression profiling for Toxoplasma, and single-nucleotide polymorphism (SNP-based genotyping of both T. gondii and P. falciparum. Expression profiling of the three clonotypic lineages dominating T. gondii populations in North America and Europe provides a first comprehensive view of the parasite transcriptome, revealing that ~49% of all annotated genes are expressed in parasite tachyzoites (the acutely lytic stage responsible for pathogenesis and 26% of genes are differentially expressed among strains. A novel design utilizing few probes provided high confidence genotyping, used here to resolve recombination points in the clonal progeny of sexual crosses. Recent sequencing of additional T. gondii isolates identifies >620 K new SNPs, including ~11 K that intersect with expression profiling probes, yielding additional markers for genotyping studies, and further validating the utility of a combined expression profiling/genotyping array design. Additional applications facilitating SNP and transcript discovery, alternative statistical methods for quantifying gene expression, etc. are also pursued at

  12. Delivery of antisense oligonucleotide to the cornea by iontophoresis. (United States)

    Berdugo, M; Valamanesh, F; Andrieu, C; Klein, C; Benezra, D; Courtois, Y; Behar-Cohen, F


    We wished to evaluate the potential of iontophoresis to promote the delivery of antisense oligonucleotides (ODN) directed at the vascular endothelial growth factor (VEGF)-R2 receptor (KDR/Flk) to the cornea of the rat eye. Fluorescence (CY5)-labeled ODNs in phosphate-buffered saline (PBS) (20 microM) were locally administered to rat eyes, and their fate within the anterior segment was studied. Thirty-four male, 5-week-old Wistar rats were used for all experiments. The rats were divided in four groups. In group I (12 rats, 12 eyes), the ODNs (20 microM) were delivered by iontophoresis (300 microA for 5 minutes) using a specially designed corneal applicator. In group II (12 rats, 12 eyes), the ODNs (20 microM) were delivered using the same applicator, but no electrical current was applied. In group III (6 rats, 6 eyes), a corneal neovascular reaction was induced prior to the application of ODNs (20 microM), and iontophoresis electrical current was delivered as for group I rats. Group IV (4 rats, 4 eyes) received ODN (60 microM) iontophoresis application (300 microA for 5 minutes) and were used for ODN integrity studies. The animals were killed 5 minutes, 90 minutes, and 24 hours after a single ODN application and studied. Topically applied ODNs using the same iontophoresis applicator but without current do not penetrate the cornea and remain confined to the superficial epithelial layer. ODNs delivered with transcorneoscleral iontophoresis penetrate into all corneal layers and are also detected in the iris. In corneas with neovascularization, ODNs were particularly localized within the vascular endothelial cells of the stroma. ODNs extracted from eye tissues 24 hours after iontophoresis remained unaltered. The iontophoresis current did not cause any detectable ocular damage under these conditions. Iontophoresis promotes the delivery of ODNs to the anterior segment of the eye, including all corneal layers. Iontophoresis of ODNs directed at VEGF-R2 may be used for the

  13. 南舟北移背景下京杭运河城市龙舟品牌赛事发展研究%On the dragon boat brand development with the background of Southern boats expanding North

    Institute of Scientific and Technical Information of China (English)



    Southern boats expanding north is the most powerful measure to prompt the development of dragon boats in China.Regarding canal city's dragon boat races as the research object,using the method of literature,field survey and interview,the dragon boat competition's situation,characteristics and existing problems are analyzed.By using the SWOT analysis theory,based on the research of its superiorities,weaknesses,opportunities and threats to develop the dragon boat brand competition,the development strategy of the dragon boat brand competition is put forward,specifically including:spread the dragon boat sports events and inherit and innovate dragon boating culture;perfect the mechanism and implement the commercial operation;rise the intensity of colleges and universities'races and make the dragon boat culture research;hold the canal city dragon boat club's super league competition,build the brand and so on,in order to further promote China's Dragon Boat Sport healthy and orderly development.%“南舟北移”成为驱动我国龙舟项目普及发展最强有力的举措。以京杭运河城市龙舟赛事为研究对象,运用文献资料、田野调查和访谈法,对当今龙舟赛事现状、特性和存在的问题进行了分析。借用SWOT分析理论,在对发展龙舟品牌赛事面临的优势、劣势、机遇和威胁进行研究的基础上,提出龙舟品牌赛事的开发策略,具体包括:推广龙舟运动,传承与创新龙舟文化;健全机制,实施商业化运作;推进高校龙舟开展力度,做好龙舟文化研究;举办运河城市龙舟俱乐部超级联赛,打造赛事品牌等,力求推动我国龙舟运动健康有序发展。

  14. Short locked nucleic acid antisense oligonucleotides potently reduce apolipoprotein B mRNA and serum cholesterol in mice and non-human primates

    DEFF Research Database (Denmark)

    Straarup, Ellen Marie; Fisker, Niels; Hedtjärn, Maj;


    the design of short antisense oligonucleotides (12- to 13-mers) that possessed high affinity and increased potency both in vitro and in vivo compared to longer oligonucleotides. The short LNA oligonucleotides were more target specific, and they exhibited the same biodistribution and tissue half......-high-density lipoprotein (non-HDL) cholesterol without increasing serum liver toxicity markers. The data presented here show that oligonucleotide length as a parameter needs to be considered in the design of antisense oligonucleotide and that potent short oligonucleotides with sufficient target affinity can be generated...

  15. Hydration-dependent dynamics of human telomeric oligonucleotides in the picosecond timescale: A neutron scattering study

    Energy Technology Data Exchange (ETDEWEB)

    Sebastiani, F.; Comez, L.; Sacchetti, F. [Dipartimento di Fisica e Geologia, Università degli Studi di Perugia, Via A. Pascoli, 06123 Perugia (Italy); CNR, Istituto Officina dei Materiali, Unità di Perugia, c/o Dipartimento di Fisica e Geologia, Università di Perugia, 06123 Perugia (Italy); Longo, M. [Dipartimento di Fisica e Geologia, Università degli Studi di Perugia, Via A. Pascoli, 06123 Perugia (Italy); Elettra—Sincrotrone Trieste, 34149 Basovizza, Trieste (Italy); Orecchini, A.; Petrillo, C.; Paciaroni, A., E-mail: [Dipartimento di Fisica e Geologia, Università degli Studi di Perugia, Via A. Pascoli, 06123 Perugia (Italy); De Francesco, A. [CNR-IOM OGG c/o Institut Laue-Langevin, 71 Avenue des Martyrs, CS20156, 38042 Grenoble Cedex 9 (France); Muthmann, M. [Jülich Centre for Neutron Science, Forschungszentrum Jülich GmbH, Outstation at Heinz Maier-Leibnitz Zentrum, Lichtenbergstrasse 1, 85747 Garching (Germany); Teixeira, S. C. M. [EPSAM, Keele University, Staffordshire ST5 5BG (United Kingdom); Institut Laue–Langevin, 71 Avenue des Martyrs, CS20156, 38042 Grenoble Cedex 9 (France)


    The dynamics of the human oligonucleotide AG{sub 3}(T{sub 2}AG{sub 3}){sub 3} has been investigated by incoherent neutron scattering in the sub-nanosecond timescale. A hydration-dependent dynamical activation of thermal fluctuations in weakly hydrated samples was found, similar to that of protein powders. The amplitudes of such thermal fluctuations were evaluated in two different exchanged wave-vector ranges, so as to single out the different contributions from intra- and inter-nucleotide dynamics. The activation energy was calculated from the temperature-dependent characteristic times of the corresponding dynamical processes. The trends of both amplitudes and activation energies support a picture where oligonucleotides possess a larger conformational flexibility than long DNA sequences. This additional flexibility, which likely results from a significant relative chain-end contribution to the average chain dynamics, could be related to the strong structural polymorphism of the investigated oligonucleotides.

  16. Repair of Thalassemic Human β -globin mRNA in Mammalian Cells by Antisense Oligonucleotides (United States)

    Sierakowska, Halina; Sambade, Maria J.; Agrawal, Sudhir; Kole, Ryszard


    In one form of β -thalassemia, a genetic blood disorder, a mutation in intron 2 of the β -globin gene (IVS2-654) causes aberrant splicing of β -globin pre-mRNA and, consequently, β -globin deficiency. Treatment of mammalian cells stably expressing the IVS2-654 human β -globin gene with antisense oligonucleotides targeted at the aberrant splice sites restored correct splicing in a dose-dependent fashion, generating correct human β -globin mRNA and polypeptide. Both products persisted for up to 72 hr posttreatment. The oligonucleotides modified splicing by a true antisense mechanism without overt unspecific effects on cell growth and splicing of other pre-mRNAs. This novel approach in which antisense oligonucleotides are used to restore rather than to down-regulate the activity of the target gene is applicable to other splicing mutants and is of potential clinical interest.

  17. Cellular uptake of antisense oligonucleotides after complexing or conjugation with cell-penetrating model peptides. (United States)

    Oehlke, J; Birth, P; Klauschenz, E; Wiesner, B; Beyermann, M; Oksche, A; Bienert, M


    The uptake by mammalian cells of phosphorothioate oligonucleotides was compared with that of their respective complexes or conjugates with cationic, cell-penetrating model peptides of varying helix-forming propensity and amphipathicity. An HPLC-based protocol for the synthesis and purification of disulfide bridged conjugates in the 10-100 nmol range was developed. Confocal laser scanning microscopy (CLSM) in combination with gel-capillary electrophoresis and laser induced fluorescence detection (GCE-LIF) revealed cytoplasmic and nuclear accumulationin all cases. The uptake differences between naked oligonucleotides and their respective peptide complexes or conjugates were generally confined to one order of magnitude. No significant influence of the structural properties of the peptide components upon cellular uptake was found. Our results question the common belief that the increased biological activity of oligonucleotides after derivatization with membrane permeable peptides may be primarily due to improved membrane translocation.

  18. Dynamics of human telomerase RNA structure revealed by antisense oligonucleotide technique. (United States)

    Vasilkova, Daria V; Azhibek, Dulat M; Zatsepin, Timofei S; Naraikina, Yulia V; Prassolov, Vladimir S; Prokofjeva, Maria M; Zvereva, Maria I; Rubtsova, Maria P


    Telomeres are the nucleoprotein complexes that cap the linear chromosome ends. Telomerase is a ribonucleoprotein that maintains telomere length in stem, embryonic and cancer cells. Somatic cells don't contain active telomerase and telomere function as mitotic clock and telomere length determines the number of cell divisions. Telomerase RNA (TER) contains the template for telomere synthesis and serves as a structural scaffold for holoenzyme assembly. We compared different oligonucleotide based methods for telomerase RNA inhibition, such as antisense oligonucleotides, knockdown by transient siRNA transfection and silencing by miRNA derived from short expressed RNA hairpin in HEK293 cells. All of these methods were applied to different TER regions. Our results revealed that CR2/CR3 domain of TER is accessible in vitro and in vivo and could serve as an optimal site for oligonucleotide-based telomerase silencing.

  19. Basic Research Objectives Reaffirmed

    Institute of Scientific and Technical Information of China (English)

    Guo Haiyan; Zhao Baohua


    @@ As a national institution for scientific research and a component of the national innovation system, CAS should and must make key contributions to the great national rejuvenation of the country. Keeping this in mind, CAS has developed four developmental targets for its basic research. This was revealed at a CAS conference on basic research held June 11-12 in Beijing.

  20. Cycles in basic innovations

    NARCIS (Netherlands)

    Groot, de E.A. (Bert); Franses, P.H.P.H.


    Basic innovations are often believed to be the drivers of economic growth. It has been widely documented that economic growth follows cyclical patterns of varying length. In this paper we examine if such patterns are also present in basic innovations. For an annual time series of count data covering

  1. Basic Science Training Program. (United States)

    Brummel, Clete

    These six learning modules were developed for Lake Michigan College's Basic Science Training Program, a workshop to develop good study skills while reviewing basic science. The first module, which was designed to provide students with the necessary skills to study efficiently, covers the following topics: time management; an overview of a study…

  2. Basic principle of superconductivity


    De Cao, Tian


    The basic principle of superconductivity is suggested in this paper. There have been two vital wrong suggestions on the basic principle, one is the relation between superconductivity and the Bose-Einstein condensation (BEC), and another is the relation between superconductivity and pseudogap.

  3. Psychosocial experiences of breast cancer survivors involved in a dragon boat program: exploring links to positive psychological growth. (United States)

    Sabiston, Catherine M; McDonough, Meghan H; Crocker, Peter R E


    This study explored psychosocial experiences of breast cancer survivors involved in dragon boat programs. Twenty women (M(age) = 58.69, SD = 6.85) were interviewed for 45-60 min about their experiences as members of survivor dragon boat teams. Interviews were analyzed using constructivist grounded theory methods. The dragon boat program facilitated social support from women with common challenges and a shared understanding of survivorship. It also provided opportunities to (re)gain a sense of personal control, develop new identities as athletes, and overcome physical challenges. Together these elements contributed to positive psychological growth and linked to the literature on posttraumatic growth. Future physical activity interventions targeting breast cancer survivors may benefit from developing strategies that share key characteristics of dragon boating.

  4. Environmental assessment of proposed dredging and disposal activities at the St. George small boat harbor, reconnaissance stage: Planning aid report (United States)

    US Fish and Wildlife Service, Department of the Interior — Types of environmental impacts possibly resulting from dredging and disposal activities at the St. George Island small boat harbor include: 1) bottom topographic and...

  5. 77 FR 38077 - Notice of Intent To Prepare an Environmental Impact Statement on a Proposed Boat-House Facility... (United States)


    ...-House Facility for Non-Motorized Boats, George Washington Memorial Parkway AGENCY: National Park Service... Impact Statement (EIS) to assess the impacts of constructing a boathouse facility and floating docks...

  6. A Study of the Response of Eastern Spiny Softshell Turtle to Boat Traffic in the Missisquoi River of Northern Vermont (United States)

    US Fish and Wildlife Service, Department of the Interior — The purpose of this study was to gauge the degree to which boat traffic in the Missisquoi River affects the basking behavior of the eastern spiny softshell turtle...

  7. An assessment of waterbird populations wintering in Port Frederick, Chichagof Island, Alaska, 19-23 February 1982: Boat surveys (United States)

    US Fish and Wildlife Service, Department of the Interior — Information is presented on the waterbird population of Port Frederick, Chichagof Island, Alaska, as revealed by small boat surveys during February 1982. The...

  8. Variance of discharge estimates sampled using acoustic Doppler current profilers from moving boats (United States)

    Garcia, Carlos M.; Tarrab, Leticia; Oberg, Kevin; Szupiany, Ricardo; Cantero, Mariano I.


    This paper presents a model for quantifying the random errors (i.e., variance) of acoustic Doppler current profiler (ADCP) discharge measurements from moving boats for different sampling times. The model focuses on the random processes in the sampled flow field and has been developed using statistical methods currently available for uncertainty analysis of velocity time series. Analysis of field data collected using ADCP from moving boats from three natural rivers of varying sizes and flow conditions shows that, even though the estimate of the integral time scale of the actual turbulent flow field is larger than the sampling interval, the integral time scale of the sampled flow field is on the order of the sampling interval. Thus, an equation for computing the variance error in discharge measurements associated with different sampling times, assuming uncorrelated flow fields is appropriate. The approach is used to help define optimal sampling strategies by choosing the exposure time required for ADCPs to accurately measure flow discharge.

  9. Use of thiolated oligonucleotides as anti-fouling diluents in electrochemical peptide-based sensors. (United States)

    McQuistan, Adam; Zaitouna, Anita J; Echeverria, Elena; Lai, Rebecca Y


    We incorporated short thiolated oligonucleotides as passivating diluents in the fabrication of electrochemical peptide-based (E-PB) sensors, with the goal of creating a negatively charged layer capable of resisting non-specific adsorption of matrix contaminants. The E-PB HIV sensors fabricated using these diluents were found to be more specific and selective, while retaining attributes similar to the sensor fabricated without these diluents. Overall, these results highlight the advantages of using oligonucleotides as anti-fouling diluents in self-assembled monolayer-based sensors.

  10. Transcriptional inhibition of the bacteriophage T7 early promoter region by oligonucleotide triple helix formation. (United States)

    Ross, C; Samuel, M; Broitman, S L


    We have identified a purine-rich triplex binding sequence overlapping a -35 transcriptional early promoter region of the bacteriophage T7. Triplex-forming oligonucleotide designed to bind this target was annealed to T7 templates and introduced into in vitro transcription systems under conditions favoring specific initiation from this promoter. These templates demonstrated significant transcriptional inhibition relative to naked genomic templates and templates mixed with non-triplex-forming oligonucleotide. It is suggested that triplex formation along this target interferes with transcriptional initiation, and this mechanism may hold potential to disrupt bacteriophage T7 early transcription in vivo.

  11. Customized oligonucleotide microchips that convert multiple genetic information to simple patterns, are portable and reusable (United States)

    Mirzabekov, Andrei; Guschin, Dmitry Y.; Chik, Valentine; Drobyshev, Aleksei; Fotin, Alexander; Yershov, Gennadiy; Lysov, Yuri


    This invention relates to using customized oligonucleotide microchips as biosensors for the detection and identification of nucleic acids specific for different genes, organisms and/or individuals in the environment, in food and in biological samples. The microchips are designed to convert multiple bits of genetic information into simpler patterns of signals that are interpreted as a unit. Because of an improved method of hybridizing oligonucleotides from samples to microchips, microchips are reusable and transportable. For field study, portable laser or bar code scanners are suitable.

  12. Oligonucleotide-templated chemical reactions: pushing the boundaries of a nature-inspired process. (United States)

    Percivalle, Claudia; Bartolo, Jean-François; Ladame, Sylvain


    Widespread in nature, oligonucleotide-templated reactions of phosphodiester bond formation have inspired chemists who are now applying this elegant strategy to the catalysis of a broad range of otherwise inefficient reactions. This review highlights the increasing diversity of chemical reactions that can be efficiently catalysed by an oligonucleotide template, using Watson-Crick base-pairing to bring both reagents in close enough proximity to react, thus increasing significantly their effective molarity. The applications of this elegant concept for nucleic acid sensing and controlled organic synthesis will also be discussed.

  13. Sequence selective naked-eye detection of DNA harnessing extension of oligonucleotide-modified nucleotides. (United States)

    Verga, Daniela; Welter, Moritz; Marx, Andreas


    DNA polymerases can efficiently and sequence selectively incorporate oligonucleotide (ODN)-modified nucleotides and the incorporated oligonucleotide strand can be employed as primer in rolling circle amplification (RCA). The effective amplification of the DNA primer by Φ29 DNA polymerase allows the sequence-selective hybridisation of the amplified strand with a G-quadruplex DNA sequence that has horse radish peroxidase-like activity. Based on these findings we develop a system that allows DNA detection with single-base resolution by naked eye.

  14. Boat Pumpout Locations, Fresh Water Boat Ramps in Rhode Island; Rs44ofr95; Public ramps and launching areas for boats and small craft on fresh water rivers streams lakes and ponds, Published in 1995, 1:24000 (1in=2000ft) scale, State of Rhode Island and Providence Plantations. (United States)

    NSGIC GIS Inventory (aka Ramona) — This Boat Pumpout Locations dataset, published at 1:24000 (1in=2000ft) scale, was produced all or in part from Hardcopy Maps information as of 1995. It is described...


    Directory of Open Access Journals (Sweden)

    Aulia Widyandari


    Full Text Available There are lot of research done to develop a patrol boat, from the modification of hull model until propulsion system equipment. For example the model ship type AMV (Advanced Marine Vehicle was developed starting from the Catamaran, Trimaran and  Pentamaran model. Everything is aimed at obtaining the ship design that has the speed and stability. In addition to achieving high-speed vessel must be equipped with propulsion (Main Power is great, that means the main engine dimensions, auxiliary equipments and fuel tanks is too large. Many Limitations of space on the ship's engine room trimaran vessel is the main obstacle in designing propulsion system. Beside that Patrol boat should have many missions speed, so propulsion system should be designed at that conditions.   Hybrid propulsion is a combination of Diesel Mechanical Propulsion (DMP with Diesel Electric Propulsion (DEP. DMP system is connected directly to the propeller shaft (or through a reduction-gear. DMP has provide more efficiency rate of 95%. While DEP is only able to provide efficiency by 85% - 89% is slightly lower than DMP, but the DEP offers many advantages such as simplicity and suitability in the rotational speed settings, control systems, engine power production Redundancy, Flexibility in the design of equipments layout in engine rooms, noise, vibration and fuel consumption efficiency which affects the lower pollution.   Design of Hybrid Propulsion system can be satisfied and achieved the Power requirements and optimally at all speed condition of patrol boat. Therefore the author made using modeling Maxsurf-11.12 software and carried out various optimization of the choice of main engine, propeller and system conditions for fast patrol boat cruise. 

  16. Development of Fluorescence Imaging Lidar for Boat-Based Coral Observation

    Directory of Open Access Journals (Sweden)

    Sasano Masahiko


    Full Text Available A fluorescence imaging lidar system installed in a boat-towable buoy has been developed for the observation of reef-building corals. Long-range fluorescent images of the sea bed can be recorded in the daytime with this system. The viability of corals is clear in these fluorescent images because of the innate fluorescent proteins. In this study, the specifications and performance of the system are shown.

  17. "We are all in the same boat" - ISEE leaders' trip to China

    Institute of Scientific and Technical Information of China (English)


    @@ The motto of Tongji University, whose name literallymeans "we are all in the same boat, and must work to-gether to reach common goals", describes the harvest ofISEE leaders' mission to China in May 2007. ISEE Presi-dent Joan Martinez-Alier, President-Elect Peter May andthe society's founder and first President, Robert Costanzaas well as ISEE member Robert Ayres, participated in aseries of events and meetings in Shanghai and Beijing witha view to build institutional collaboration.

  18. The Intrusion of Engine Exhaust into the Passenger Areas of Recreational Power Boats (United States)


    Annapolis, Maryland. The test boat was a 34-foot Silverton sedan cruiser equipped with twin 302 CID engines with exhaust exits located on each side of the...40.60 30.14 1.42 09:36:31 51.80 41.90 30.14 2.74 -001 -000 -@00 @00 test oeat: 361 Silverton Sedan Cruiser 19:27:01 51.30 42.70 30.14 1.53 *forever

  19. Studies on the Syntheses and Properties of 5'-Branched-sugar Isonucleosides and the Related Oligonucleotides

    Institute of Scientific and Technical Information of China (English)

    TianXiaobing; ZhangLihe; MinJimei


    The chemistry of nucleosides and oligonucleotides is an actively investigated field in the search for new drugs. Thesyntheses and the properties of isonucleosides and oligonucleotides have been investigated to improve their stability,antitumor and antiviral activities, and to reduce their toxicity.

  20. Synthesis, Dynamic Combinatorial Chemistry, and PCR Amplification of 3'-5' and 3'-6' Disulfide-linked Oligonucleotides

    DEFF Research Database (Denmark)

    Hansen, Dennis Jul; Manuguerra, Ilenia; Kjelstrup, Michael Brøndum;


    Disulfide dithymidines linked 3'-5' or 3'-6' were synthesized and incorporated into oligonucleotides through a combined phosphotriester and phosphoramidite solid-phase oligonucleotide synthesis approach. The disulfide links are cleaved and formed reversibly in the presence of thiols and oligonucl...

  1. Oligonucleotides Containing Aminated 2'-Amino-LNA Nucleotides: Synthesis and Strong Binding to Complementary DNA and RNA. (United States)

    Lou, Chenguang; Samuelsen, Simone V; Christensen, Niels Johan; Vester, Birte; Wengel, Jesper


    Mono- and diaminated 2'-amino-LNA monomers were synthesized and introduced into oligonucleotides. Each modification imparts significant stabilization of nucleic acid duplexes and triplexes, excellent sequence selectivity, and significant nuclease resistance. Molecular modeling suggested that structural stabilization occurs via intrastrand electrostatic attraction between the protonated amino groups of the aminated 2'-amino-LNA monomers and the host oligonucleotide backbone.

  2. Isolation and application of Gordonia sp. JC11 for removal of boat lubricants. (United States)

    Chanthamalee, Jirapat; Luepromchai, Ekawan


    Boat lubricants are continuously released into the marine environment and thereby cause chronic oil pollution. This study aims to isolate lubricant-degrading microorganisms from Thai coastal areas as well as to apply a selected strain for removal of boat lubricants. Ten microorganisms in the genera of Gordonia, Microbacterium, Acinetobacter, Pseudomonas, Brucella, Enterococcus and Candida were initially isolated by crude oil enrichment culture techniques. The lubricant-removal activity of these isolates was investigated with mineral-based lubricants that had been manufactured for the 4-stroke diesel engines of fishing boats. Gordonia sp. JC11, the most effective strain was able to degrade 25-55% of 1,000 mg L(-1) total hydrocarbons in six tested lubricants, while only 0-15% of the lubricants was abiotically removed. The bacterium had many characteristics that promoted lubricant degradation such as hydrocarbon utilization ability, emulsification activity and cell surface hydrophobicity. For bioaugmentation treatment of lubricant contaminated seawater, the inoculum of Gordonia sp. JC11 was prepared by immobilizing the bacterium on polyurethane foam (PUF). PUF-immobilized Gordonia sp. JC11 was able to remove 42-56% of 100-1,000 mg L(-1) waste lubricant No. 2 within 5 days. This lubricant removal efficiency was higher than those of free cells and PUF without bacterial cells. The bioaugmentation treatment significantly increased the number of lubricant-degrading microorganisms in the fishery port seawater microcosm and resulted in rapid removal of waste lubricant No. 2.

  3. Basic stress analysis

    CERN Document Server

    Iremonger, M J


    BASIC Stress Analysis aims to help students to become proficient at BASIC programming by actually using it in an important engineering subject. It also enables the student to use computing as a means of learning stress analysis because writing a program is analogous to teaching-it is necessary to understand the subject matter. The book begins by introducing the BASIC approach and the concept of stress analysis at first- and second-year undergraduate level. Subsequent chapters contain a summary of relevant theory, worked examples containing computer programs, and a set of problems. Topics c

  4. Quantum electronics basic theory

    CERN Document Server

    Fain, V M; Sanders, J H


    Quantum Electronics, Volume 1: Basic Theory is a condensed and generalized description of the many research and rapid progress done on the subject. It is translated from the Russian language. The volume describes the basic theory of quantum electronics, and shows how the concepts and equations followed in quantum electronics arise from the basic principles of theoretical physics. The book then briefly discusses the interaction of an electromagnetic field with matter. The text also covers the quantum theory of relaxation process when a quantum system approaches an equilibrium state, and explai

  5. Boat ramp locations within the Columbia River Basin with associated recreational use, water quality measurements, and risk assessment data for zebra and quagga mussels (United States)

    Hardiman, Jill M.; Holmberg, Glen S.; Elder, Nancy E.


    Aquatic invasive species are often transported between water bodies on boats or boat trailers, thus they are considered one of the primary vectors for new introductions of invasive species to a water body. This data set contains geographic positioning system locational data for boater access points, use data (i.e. recreational, fishing), water quality measurements (e.g. calcium concentrations, pH), risk assessment data, and other physical attributes (i.e. size, elevation) where available within the Columbia and Snake Rivers and throughout the Columbia River Basin. This work builds on an earlier body of work by Wells et al. 2011, Prioritizing Zebra and Quagga Mussel Monitoring in the Columbia River Basin (PDF link below), which provided much of the initial water quality, use information, and risk assessment data (categorical values for the risk of introduction and the risk of establishment). Updated information has been added by collecting additional data on use of water bodies, as well as combined categorical ranking methodology for identifying water bodies that may be high risk for both introduction and establishment of zebra and quagga mussels. This data is also related to a regional effort to contribute to the coordination of monitoring efforts for early detection of zebra and quagga mussels in the context of risk assessment data (CRBAIS weblink below). Data sets provided here include a service definition file, provided with a few reference layers within the region for viewing and an online map ( with some query options, a GIS shapefile (Child Item), and a tabular data set (csv file; Child Item). It is recommended that all users of this data thoroughly read the metadata files for data definitions, sources, and data limitations. It is recommended to use the online map link ( for a quick view of the data set and some basic query options.  Once on the ArcGIS online map, to view all the data layers click the show

  6. Video Screen Capture Basics (United States)

    Dunbar, Laura


    This article is an introduction to video screen capture. Basic information of two software programs, QuickTime for Mac and BlueBerry Flashback Express for PC, are also discussed. Practical applications for video screen capture are given.

  7. HIV Treatment: The Basics (United States)

    HIV Treatment HIV Treatment: The Basics (Last updated 2/24/2017; last reviewed 2/24/2017) Key Points Antiretroviral therapy (ART) ... reduces the risk of HIV transmission . How do HIV medicines work? HIV attacks and destroys the infection- ...

  8. Kidney Disease Basics (United States)

    ... Links Take the first step Alternate Language URL Kidney Disease Basics Page Content Your kidneys filter extra ... blood pressure are the most common causes of kidney disease. ​These conditions can slowly damage the kidneys ...

  9. Health Literacy Basics (United States)

    ... have the capacity to obtain, process, and understand basic health information and services needed to make appropriate health decisions. 1 Health literacy is dependent on individual and systemic factors: Communication skills of lay persons and professionals Lay and professional ...

  10. Basic Financial Accounting

    DEFF Research Database (Denmark)

    Wiborg, Karsten

    This textbook on Basic Financial Accounting is targeted students in the economics studies at universities and business colleges having an introductory subject in the external dimension of the company's economic reporting, including bookkeeping, etc. The book includes the following subjects...

  11. Basic Concurrency Theory

    DEFF Research Database (Denmark)

    Løvengreen, Hans Henrik


    In this set of notes, we present some of the basic theory underlying the discipline of programming with concurrent processes/threads. The notes are intended to supplement a standard textbook on concurrent programming.......In this set of notes, we present some of the basic theory underlying the discipline of programming with concurrent processes/threads. The notes are intended to supplement a standard textbook on concurrent programming....

  12. Emissions in the exhaust of fishing boats after adding viscous agents into fuel oils. (United States)

    Hsieh, Lien-Te; Shih, Shun-I; Lin, Sheng-Lun; Yang, Tsun-Lirng; Wu, Tser-Son; Hung, Chung-Hsien


    In order to avoid the illegal use of fishing boat fuel A (FBFA) by traveling diesel vehicles (TDVs) in Taiwan, alternatives that are easily distinguished from premium diesel fuel (PDF) were prepared to evaluate their suitability. Two new ingredients, pyrolysis fuel oil (PFO) and residue of desulfurization unit (RDS), were added into FBFA and formed PFO0.5 and RDS0.5, respectively. Along with FBFA, these three fuels were analyzed for their chemical and physical properties. Furthermore, they were used by three fishing boats with different sizes, output powers, and weights. The engine performances and pollutant emissions were examined and monitored. Experimental results show that there are significant differences in appearance between PDF and the two new blended fuels (PFO0.5 and RDS0.5), and thus misuse or illegal use of FBFA could be substantially reduced. The fuel consumption, which is negatively related to the heating value of fuels, is in order of FBFAfishing boats, using RDS0.5 resulted in a decrease in CO and NO(x) emissions, while the PM emission factors (g bhp(-1) h(-1) and g L(-1)-fuel) were reduced by approximately 36% and 33%, respectively. Owing to the higher total aromatic content in PFO0.5 and RDS0.5, total-PAH concentrations in the exhausts from the three fishing boats using PFO0.5 and RDS0.5 were slightly (1.2 and 1.1 times, respectively) higher than for those using FBFA. Nevertheless, the estimated total BaP(eq) from the three fishing boats using RDS0.5 was 27.5, 19.5, and 8.25% lower than those using FBFA. With using PFO0.5, they were totally different, at 23.5, 2.79, and 2.58% higher. With regard to looking different to PDF, RDS0.5 is superior to PFO0.5, and is thus recommended as a better alternative to FBFA, particularly because it can help lower more emissions of CO, NO(x), PM and BaP(eq).

  13. Neutron Star Kicks by the Gravitational Tug-boat Mechanism in Asymmetric Supernova Explosions: Progenitor and Explosion Dependence (United States)

    Janka, Hans-Thomas


    Asymmetric mass ejection in the early phase of supernova (SN) explosions can impart a kick velocity to the new-born neutron star (NS). For neutrino-driven explosions the NS acceleration has been shown to be mainly caused by the gravitational attraction of the anisotropically expelled inner ejecta, while hydrodynamic forces contribute on a subdominant level, and asymmetric neutrino emission plays only a secondary role. Two- and three-dimensional hydrodynamic simulations have demonstrated that this gravitational tug-boat mechanism can explain the observed space velocities of young NSs up to more than 1000 km s‑1. Here, we discuss how the NS kick depends on the energy, ejecta mass, and asymmetry of the SN explosion, and what role the compactness of the pre-collapse stellar core plays for the momentum transfer to the NS. We also provide simple analytic expressions for the NS velocity in terms of these quantities. Referring to results of hydrodynamic simulations in the literature, we argue why, within the discussed scenario of NS acceleration, electron-capture SNe, low-mass Fe-core SNe, and ultra-stripped SNe can be expected to have considerably lower intrinsic NS kicks than core-collapse SNe of massive stellar cores. Our basic arguments also remain valid if progenitor stars possess large-scale asymmetries in their convective silicon and oxygen burning layers. Possible scenarios for spin-kick alignment are sketched. Much of our discussion stays on a conceptual and qualitative level, and more work is necessary on the numerical modeling side to determine the dependences of involved parameters, whose prescriptions will be needed for recipes that can be used to better describe NS kicks in binary evolution and population synthesis studies.

  14. Comparison of small molecules and oligonucleotides that target a toxic, non-coding RNA. (United States)

    Costales, Matthew G; Rzuczek, Suzanne G; Disney, Matthew D


    Potential RNA targets for chemical probes and therapeutic modalities are pervasive in the transcriptome. Oligonucleotide-based therapeutics are commonly used to target RNA sequence. Small molecules are emerging as a modality to target RNA structures selectively, but their development is still in its infancy. In this work, we compare the activity of oligonucleotides and several classes of small molecules that target the non-coding r(CCUG) repeat expansion (r(CCUG)(exp)) that causes myotonic dystrophy type 2 (DM2), an incurable disease that is the second-most common cause of adult onset muscular dystrophy. Small molecule types investigated include monomers, dimers, and multivalent compounds synthesized on-site by using RNA-templated click chemistry. Oligonucleotides investigated include phosphorothioates that cleave their target and vivo-morpholinos that modulate target RNA activity via binding. We show that compounds assembled on-site that recognize structure have the highest potencies amongst small molecules and are similar in potency to a vivo-morpholino modified oligonucleotide that targets sequence. These studies are likely to impact the design of therapeutic modalities targeting other repeats expansions that cause fragile X syndrome and amyotrophic lateral sclerosis, for example.

  15. A triple-helix forming oligonucleotide targeting genomic DNA fails to induce mutation. (United States)

    Reshat, Reshat; Priestley, Catherine C; Gooderham, Nigel J


    Purine tracts in duplex DNA can bind oligonucleotide strands in a sequence specific manner to form triple-helix structures. Triple-helix forming oligonucleotides (TFOs) targeting supFG1 constructs have previously been shown to be mutagenic raising safety concerns for oligonucleotide-based pharmaceuticals. We have engineered a TFO, TFO27, to target the genomic Hypoxanthine-guanine phosphoribosyltransferase (HPRT) locus to define the mutagenic potential of such structures at genomic DNA. We report that TFO27 was resistant to nuclease degradation and readily binds to its target motif in a cell free system. Contrary to previous studies using the supFG1 reporter construct, TFO27 failed to induce mutation within the genomic HPRT locus. We suggest that it is possible that previous reports of triplex-mediated mutation using the supFG1 reporter construct could be confounded by DNA quadruplex formation. Although the present study indicates that a TFO targeting a genomic locus lacks mutagenic activity, it is unclear if this finding can be generalised to all TFOs and their targets. For the present, we suggest that it is prudent to avoid large purine stretches in oligonucleotide pharmaceutical design to minimise concern regarding off-target genotoxicity.

  16. Effect of iontophoresis on the in vitro trans-scleral transport of three single stranded oligonucleotides. (United States)

    Pescina, Silvia; Antopolsky, Maxim; Santi, Patrizia; Nicoli, Sara; Murtomäki, Lasse


    Oligonucleotides represent a subject of clinical interest due to their potential ability to treat several diseases, including those affecting the posterior segment of the eye. Unfortunately, therapeutic oligonucleotides are currently administered by means of highly invasive approaches, such as intravitreal injections. The aim of the present work was to study in vitro, across isolated bovine sclera, the effect of iontophoresis on the transport of three single stranded oligonucleotides (ssDNA), 12-, 24- and 36-mer, selected as reference compounds in view of a non-invasive drug delivery to the back of the eye. All the three sequences were able to cross bovine sclera in vitro without iontophoresis. When anodal iontophoresis was applied, no change in flux was observed, while in the presence of cathodal iontophoresis the permeability coefficients increased four-fold compared to passive conditions. This behavior can be ascribed to the electrorepulsive mechanism, due to the negative charge of the nucleic acid backbone. It was also observed that the molecular weights of the three sequences did not affect trans-scleral transport, neither in passive, nor in current assisted permeation. Furthermore, increasing the current intensity from 1.75 mA to 3 mA, no effect on the trans-scleral transport of the 24-mer was noticed. Although preliminary, the results demonstrate that cathodal iontophoresis enhances trans-scleral transport of single stranded oligonucleotides and suggest its use as a novel non-invasive approach for the treatment of diseases affecting the posterior segment of the eye.

  17. In situ entry of oligonucleotides into brain cells can occur through a nucleic acid channel

    NARCIS (Netherlands)

    Shi, Fuxin; Gounko, Natasha V.; Wang, Xiaoqin; Ronken, Eric; Hoekstra, Dick


    Brain tissue has become a challenging therapeutic target, in part because of failure of conventional treatments of brain tumors and a gradually increasing number of neurodegenerative diseases. Because antisense oligonucleotides are readily internalized by neuronal cells in culture, these compounds c

  18. A new achiral reagent for the incorporation of multiple amino groups into oligonucleotides

    DEFF Research Database (Denmark)

    Behrens, Carsten; Petersen, Kenneth H.; Egholm, Michael;


    The synthesis of a new functionalized achiral linker reagent (10) for the incorporation of multiple primary amino groups into oligonucleotides is described. The linker reagent is compatible with conventional DNA-synthesis following the phosphoramidite methodology, and the linker can be incorporat...

  19. Application of decoy oligonucleotides as novel therapeutic strategy: a contemporary overview. (United States)

    Ahmad, Mohammad Zaki; Akhter, Sohail; Mallik, Neha; Anwar, Mohammad; Tabassum, Wajda; Ahmad, Farhan Jalees


    Molecular therapy is emerging as a potential strategy for the treatment of many diseases. Correct regulation of gene expression is essential for both, to normal development and proper functioning of the all the organisms. Even after four decades of intensive research, it is still a major problem from regulatory and technical point of view, to replace defective genes. The technology of decoy oligonucleotides has received considerable attention to treat and cure a variety of diseases and abnormal physiological conditions, because they provide a rational way to design and selective regulation of a specific gene expression. Decoy oligonucleotides are widely used as inhibitors of specific gene expression because they can offer exciting possibility of expression and blocking of a particular gene without any changes in the functions of other genes. Advances in the decoy oligonucleotides are rapidly paving the way to new insights into the origin and treatment of inflammatory, cancer and/or other immune disorders. The review covers the progress achieved towards the development of decoy oligonucleotides as a potential strategy in a new class of molecular therapy.

  20. High-frequency genome editing using ssDNA oligonucleotides with zinc-finger nucleases

    DEFF Research Database (Denmark)

    Chen, Fuqiang; Pruett-Miller, Shondra M; Huang, Yuping


    Zinc-finger nucleases (ZFNs) have enabled highly efficient gene targeting in multiple cell types and organisms. Here we describe methods for using simple ssDNA oligonucleotides in tandem with ZFNs to efficiently produce human cell lines with three distinct genetic outcomes: (i) targeted point...

  1. Pd0-Catalyzed Methyl Transfer on Nucleosides and Oligonucleotides, Envisaged as a PET Tracer

    Directory of Open Access Journals (Sweden)

    Eric Fouquet


    Full Text Available The methyl transfer reaction from activated monomethyltin, via a modified Stille coupling reaction, was studied under “ligandless” conditions on fully deprotected 5'-modified nucleosides and one dinucleotide. The reaction was optimized to proceed in a few minutes and quantitative yield, even under dilute conditions, thus affording a rapid and efficient new method for oligonucleotide labelling with carbon-11.

  2. Refinement of antisense oligonucleotide mediated exon skipping as therapy for Duchenne muscular dystrophy

    NARCIS (Netherlands)

    Heemskerk, Johannes Antonius


    In recent years, modulation of mRNA has emerged as a promising therapeutic tool. For instance, in the field of neuromuscular disorders therapeutic strategies are being developed for several diseases, including antisense oligonucleotide (AON) mediated exon skipping for Duchenne Muscular Dystrophy (DM

  3. Stable gene targeting in human cells using single-strand oligonucleotides with modified bases.

    Directory of Open Access Journals (Sweden)

    Xavier Rios

    Full Text Available Recent advances allow multiplexed genome engineering in E. coli, employing easily designed oligonucleotides to edit multiple loci simultaneously. A similar technology in human cells would greatly expedite functional genomics, both by enhancing our ability to test how individual variants such as single nucleotide polymorphisms (SNPs are related to specific phenotypes, and potentially allowing simultaneous mutation of multiple loci. However, oligo-mediated targeting of human cells is currently limited by low targeting efficiencies and low survival of modified cells. Using a HeLa-based EGFP-rescue reporter system we show that use of modified base analogs can increase targeting efficiency, in part by avoiding the mismatch repair machinery. We investigate the effects of oligonucleotide toxicity and find a strong correlation between the number of phosphorothioate bonds and toxicity. Stably EGFP-corrected cells were generated at a frequency of ~0.05% with an optimized oligonucleotide design combining modified bases and reduced number of phosphorothioate bonds. We provide evidence from comparative RNA-seq analysis suggesting cellular immunity induced by the oligonucleotides might contribute to the low viability of oligo-corrected cells. Further optimization of this method should allow rapid and scalable genome engineering in human cells.

  4. Optimization of single-base-pair mismatch discrimination in oligonucleotide microarrays

    NARCIS (Netherlands)

    Urakawa, H.; Fantroussi, El S.; Smidt, H.; Smoot, J.C.; Tribou, E.H.; Kelly, J.J.; Noble, P.A.; Stahl, D.A.


    The discrimination between perfect-match and single-base-pair-mismatched nucleic acid duplexes was investigated by using oligonucleotide DNA microarrays and nonequilibrium dissociation rates (melting profiles). DNA and RNA versions of two synthetic targets corresponding to the 16S rRNA sequences of

  5. Cellular Internalization of Therapeutic Oligonucleotides by Peptide Amphiphile Nanofibers and Nanospheres. (United States)

    Mumcuoglu, Didem; Sardan Ekiz, Melis; Gunay, Gokhan; Tekinay, Turgay; Tekinay, Ayse B; Guler, Mustafa O


    Oligonucleotides are promising drug candidates due to the exceptionally high specificity they exhibit toward their target DNA and RNA sequences. However, their poor pharmacokinetic and pharmacodynamic properties, in conjunction with problems associated with their internalization by cells, necessitates their delivery through specialized carrier systems for efficient therapy. Here, we investigate the effects of carrier morphology on the cellular internalization mechanisms of oligonucleotides by using self-assembled fibrous or spherical peptide nanostructures. Size and geometry were both found to be important parameters for the oligonucleotide internalization process; direct penetration was determined to be the major mechanism for the internalization of nanosphere carriers, whereas nanofibers were internalized by clathrin- and dynamin-dependent endocytosis pathways. We further showed that glucose conjugation to carrier nanosystems improved cellular internalization in cancer cells due to the enhanced glucose metabolism associated with oncogenesis, and the internalization of the glucose-conjugated peptide/oligonucleotide complexes was found to be dependent on glucose transporters present on the surface of the cell membrane.

  6. Methods for the preparation of large quantities of complex single-stranded oligonucleotide libraries. (United States)

    Murgha, Yusuf E; Rouillard, Jean-Marie; Gulari, Erdogan


    Custom-defined oligonucleotide collections have a broad range of applications in fields of synthetic biology, targeted sequencing, and cytogenetics. Also, they are used to encode information for technologies like RNA interference, protein engineering and DNA-encoded libraries. High-throughput parallel DNA synthesis technologies developed for the manufacture of DNA microarrays can produce libraries of large numbers of different oligonucleotides, but in very limited amounts. Here, we compare three approaches to prepare large quantities of single-stranded oligonucleotide libraries derived from microarray synthesized collections. The first approach, alkaline melting of double-stranded PCR amplified libraries with a biotinylated strand captured on streptavidin coated magnetic beads results in little or no non-biotinylated ssDNA. The second method wherein the phosphorylated strand of PCR amplified libraries is nucleolyticaly hydrolyzed is recommended when small amounts of libraries are needed. The third method combining in vitro transcription of PCR amplified libraries to reverse transcription of the RNA product into single-stranded cDNA is our recommended method to produce large amounts of oligonucleotide libraries. Finally, we propose a method to remove any primer binding sequences introduced during library amplification.

  7. Precise construction of oligonucleotide-Fab fragment conjugate for homogeneous immunoassay using HaloTag technology. (United States)

    Päkkilä, Henna; Peltomaa, Riikka; Lamminmäki, Urpo; Soukka, Tero


    The use of oligonucleotide-protein conjugates enables the development of novel types of bioanalytical assays. However, convenient methods for producing covalent and stoichiometric oligonucleotide-protein conjugates are still rare. Here we demonstrate, for the first time, covalent conjugation of DNA oligonucleotide to Fab fragments with a 1:1 ratio using HaloTag self-labeling technology. The oligonucleotide coupling was carried out while the Fab was attached to protein G matrix, thereby enabling straightforward production of covalent conjugates. Furthermore, it allowed convenient purification of the product because the unreacted components were easily removed before the elution of the high-purity conjugate. The prepared conjugate was employed in a homogeneous immunoassay where prostate-specific antigen was used as a model analyte. Switchable lanthanide luminescence was used for detection, and the obtained limit of detection was 0.27 ng/ml. In the future, the developed method for covalent conjugation and successive purification in protein G column could also be applied for introducing other kinds of modifications to Fab fragments in a simple and site-specific manner.

  8. Rapid identification of bacteria in blood cultures by using fluorescently labeled oligonucleotide probes

    NARCIS (Netherlands)

    Jansen, GJ; Mooibroek, M; Idema, J; Harmsen, HJM; Welling, GW; Degener, JE


    The applicability of whole-cell hybridization for the identification of pathogenic bacteria in blood from septic patients was examined. Oligonucleotide probes, fluorescently labeled with fluorescein isothiocyanate, directed against the variable regions of the 16S rRNAs of the following bacterial spe

  9. PCSK9 LNA antisense oligonucleotides induce sustained reduction of LDL cholesterol in nonhuman primates. (United States)

    Lindholm, Marie W; Elmén, Joacim; Fisker, Niels; Hansen, Henrik F; Persson, Robert; Møller, Marianne R; Rosenbohm, Christoph; Ørum, Henrik; Straarup, Ellen M; Koch, Troels


    Proprotein convertase subtilisin/kexin type 9 (PCSK9) has emerged as a therapeutic target for the reduction of low-density lipoprotein cholesterol (LDL-C). PCSK9 increases the degradation of the LDL receptor, resulting in high LDL-C in individuals with high PCSK9 activity. Here, we show that two locked nucleic acid (LNA) antisense oligonucleotides targeting PCSK9 produce sustained reduction of LDL-C in nonhuman primates after a loading dose (20 mg/kg) and four weekly maintenance doses (5 mg/kg). PCSK9 messenger RNA (mRNA) and serum PCSK9 protein were reduced by 85% which resulted in a 50% reduction in circulating LDL-C. Serum total cholesterol (TC) levels were reduced to the same extent as LDL-C with no reduction in high-density lipoprotein levels, demonstrating a specific pharmacological effect on LDL-C. The reduction in hepatic PCSK9 mRNA correlated with liver LNA oligonucleotide content. This verified that anti-PCSK9 LNA oligonucleotides regulated LDL-C through an antisense mechanism. The compounds were well tolerated with no observed effects on toxicological parameters (liver and kidney histology, alanine aminotransferase, aspartate aminotransferase, urea, and creatinine). The pharmacologic evidence and initial safety profile of the compounds used in this study indicate that LNA antisense oligonucleotides targeting PCSK9 provide a viable therapeutic strategy and are potential complements to statins in managing high LDL-C.

  10. Design and analysis of effects of triplet repeat oligonucleotides in cell models for myotonic dystrophy

    NARCIS (Netherlands)

    Gonzalez-Barriga, A.; Mulders, S.A.M.; Giessen, J. van der; Hooijer, J.D.; Bijl, S.; Kessel, I.D.G. van; Beers, J. van; Deutekom, J.C. van; Fransen, J.A.M.; Wieringa, B.; Wansink, D.G.


    Myotonic dystrophy type 1 (DM1) is caused by DM protein kinase (DMPK) transcripts containing an expanded (CUG)n repeat. Antisense oligonucleotide (AON)-mediated suppression of these mutant RNAs is considered a promising therapeutic strategy for this severe disorder. Earlier, we identified a 2'-O-met

  11. Negative electrospray ionization mass spectrometry of synthetic and chemically modified oligonucleotides

    NARCIS (Netherlands)

    Potier, N.; Van Dorsselaer, A.; Cordier, Y.; Roch, O.; Bischoff, Rainer


    We report here on the analysis of synthetic oligonucleotides by electrospray ionization mass spectrometry (ESI-MS). After intensive removal of salt ions (especially sodium cations), negative ion mass spectra, allowing mass measurement with an accuracy of 0.01%, were obtained on several oligonucleoti

  12. Methods for the preparation of large quantities of complex single-stranded oligonucleotide libraries.

    Directory of Open Access Journals (Sweden)

    Yusuf E Murgha

    Full Text Available Custom-defined oligonucleotide collections have a broad range of applications in fields of synthetic biology, targeted sequencing, and cytogenetics. Also, they are used to encode information for technologies like RNA interference, protein engineering and DNA-encoded libraries. High-throughput parallel DNA synthesis technologies developed for the manufacture of DNA microarrays can produce libraries of large numbers of different oligonucleotides, but in very limited amounts. Here, we compare three approaches to prepare large quantities of single-stranded oligonucleotide libraries derived from microarray synthesized collections. The first approach, alkaline melting of double-stranded PCR amplified libraries with a biotinylated strand captured on streptavidin coated magnetic beads results in little or no non-biotinylated ssDNA. The second method wherein the phosphorylated strand of PCR amplified libraries is nucleolyticaly hydrolyzed is recommended when small amounts of libraries are needed. The third method combining in vitro transcription of PCR amplified libraries to reverse transcription of the RNA product into single-stranded cDNA is our recommended method to produce large amounts of oligonucleotide libraries. Finally, we propose a method to remove any primer binding sequences introduced during library amplification.

  13. Obstructive Effects of Ultrasonic Microbubble Intensifier on CHG-5 Cell with Survivin Antisense Oligonucleotides Transfection

    Institute of Scientific and Technical Information of China (English)

    CAO Hong-ying; CAO You-de; WANG Zhi-gang; LI Pan


    Objective:To study the effects on human glioma cell line CHG-5 by ultrasonic microbubble intensifier with survivin antisense oligonucleotides (ASODN)transfection. Methods: Antisense oligonucleotides targeting survivin mRNA was designed and synthesized.Four regimen groups were designed,group A:survivin antisense oligonucleotides transfected with ultrasonic microbubble intensifier combined with ultrasound irradiation,group B: survivin antisense oligonucleotides transfected with lipofectamine combined with ultrasound irradiation,group C:survivin antisense oligonucelotides with lipofectamine D:blank control.The expression changes of surviving protein were measured by immunohistochemical staining and Western blotting,and MTr assay was used to measure the changes of proliferation.Results:Survivin protein expression in group A was decreased significantly in human glioma cell line CHG-5 than other groups(P<0.05),and the proliferating rate of CHG-5 in group A was also significantly inhibited(P<0.05).Conclusion:Ultrasonic microbubble intensifier transfection combined with ultrasound irradiation is a promising method in gene transfection effectively and noninvasively.

  14. An oligonucleotide-tagged microarray for routine diagnostics of colon cancer by genotyping KRAS mutations

    DEFF Research Database (Denmark)

    Liu, Yuliang; Guðnason, Haukur; Li, Yiping


    or spiked fecal samples. The immobilized tag-probes were stable under multiple thermal cycling treatments, allowing re-use of the tag-microarray and further optimization to solid PCR. Our results demonstrated that a novel oligonucleotide-tagged microarray system has been developed which would be suitable...

  15. Oligonucleotide array outperforms SNP array on formalin-fixed paraffin-embedded clinical samples. (United States)

    Nasri, Soroush; Anjomshoaa, Ahmad; Song, Sarah; Guilford, Parry; McNoe, Les; Black, Michael; Phillips, Vicky; Reeve, Anthony; Humar, Bostjan


    Compromised quality of formalin-fixed paraffin-embedded (FFPE)-derived DNA has compounded the use of archival specimens for array-based genomic studies. Recent technological advances have led to first successes in this field; however, there is currently no general agreement on the most suitable platform for the array-based analysis of FFPE DNA. In this study, FFPE and matched fresh-frozen (FF) specimens were separately analyzed with Affymetrix single nucleotide polymorphism (SNP) 6.0 and Agilent 4x44K oligonucleotide arrays to compare the genomic profiles from the two tissue sources and to assess the relative performance of the two platforms on FFPE material. Genomic DNA was extracted from matched FFPE-FF pairs of normal intestinal epithelium from four patients and were applied to the SNP and oligonucleotide platforms according to the manufacturer-recommended protocols. On the Affymetrix platform, a substantial increase in apparent copy number alterations was observed in all FFPE tissues relative to their matched FF counterparts. In contrast, FFPE and matched FF genomic profiles obtained via the Agilent platform were very similar. Both the SNP and the oligonucleotide platform performed comparably on FF material. This study demonstrates that Agilent oligonucleotide array comparative genomic hybridization generates reliable results from FFPE extracted DNA, whereas the Affymetrix SNP-based array seems less suitable for the analysis of FFPE material.

  16. Tetrahedron-structured DNA and functional oligonucleotide for construction of an electrochemical DNA-based biosensor. (United States)

    Bu, Nan-Nan; Tang, Chun-Xia; He, Xi-Wen; Yin, Xue-Bo


    Tetrahedron-structured DNA (ts-DNA) in combination with a functionalized oligonucleotide was used to develop a "turn-on" biosensor for Hg(2+) ions. The ts-DNA provided an improved sensitivity and was used to block the active sites.

  17. An oral oligonucleotide delivery system based on a thiolated polymer: Development and in vitro evaluation. (United States)

    Martien, Ronny; Hoyer, Herbert; Perera, Glen; Schnürch, Andreas Bernkop


    The purpose of this study was to develop and evaluate an oral oligonucleotide delivery system based on a thiolated polymer/reduced glutathione (GSH) system providing a protective effect toward nucleases and permeation enhancement. A polycarbophil-cysteine conjugate (PCP-Cys) was synthesized. Enzymatic degradation of a model oligonucleotide by DNase I and within freshly collected intestinal fluid was investigated in the absence and presence of PCP-Cys. Permeation studies with PCP-Cys/GSH versus control were performed in vitro on Caco-2 cell monolayers and ex vivo on rat intestinal mucosa. PCP-Cys displayed 223 ± 13.8 μmol thiol groups per gram polymer. After 4h, 61% of the free oligonucleotides were degraded by DNase I and 80% within intestinal fluid. In contrast, less than 41% (DNase I) and 60% (intestinal fluid) were degraded in the presence of 0.02% (m/v) PCP-Cys. Permeation studies revealed an 8-fold (Caco-2) and 10-fold (intestinal mucosa) increase in apparent permeability compared to buffer control. Hence, this PCP-Cys/GSH system might be a promising tool for the oral administration of oligonucleotides as it allows a significant protection toward degrading enzymes and facilitates their transport across intestinal membranes.

  18. Mechanical properties and fabrication of small boat using woven glass/sugar palm fibres reinforced unsaturated polyester hybrid composite (United States)

    Misri, S.; Leman, Z.; Sapuan, S. M.; Ishak, M. R.


    In recent years, sugar palm fibre has been found to have great potential to be used as fibre reinforcement in polymer matrix composites. This research investigates the mechanical properties of woven glass/sugar palm fibres reinforced unsaturated polyester hybrid composite. The composite specimens made of different layer of fibres such as strand mat, natural and hand woven of sugar palm fibres. The composites were fabricated using a compression moulding technique. The tensile and impact test was carried out in accordance to ASTM 5083 and ASTM D256 standard. The fibre glass boat is a familiar material used in boat industry. A lot of research on fabrication process such as lay-up, vacuum infusion mould and resin transfer mould has been conducted. Hybrid material of sugar palm fibre and fibre glass was used in fabricating the boat. This research investigates the method selection for fabrication of small boat application of natural fibre composites. The composite specimens made of different layer of fibres; woven glass fibre, strand mat, natural and hand woven of woven sugar palm fibres were prepared. The small boat were fabricated using a compression moulding and lay up technique. The results of the experiment showed that the tensile strength, tensile modulus, elongation at break value and impact strength were higher than the natural woven sugar palm fibre. The best method for fabricating the small boat was compression moulding technique. As a general conclusion, the usage of glass fibre had improved the tensile properties sugar palm fibre composites and compression moulding technique is suitable to be used in making a small boat application of natural fibre composites.

  19. Short G-rich oligonucleotides as a potential therapeutic for Huntington's Disease

    Directory of Open Access Journals (Sweden)

    Parekh-Olmedo Hetal


    Full Text Available Abstract Background Huntington's Disease (HD is an inherited autosomal dominant genetic disorder in which neuronal tissue degenerates. The pathogenesis of the disease appears to center on the development of protein aggregates that arise initially from the misfolding of the mutant HD protein. Mutant huntingtin (Htt is produced by HD genes that contain an increased number of glutamine codons within the first exon and this expansion leads to the production of a protein that misfolds. Recent studies suggest that mutant Htt can nucleate protein aggregation and interfere with a multitude of normal cellular functions. Results As such, efforts to find a therapy for HD have focused on agents that disrupt or block the mutant Htt aggregation pathway. Here, we report that short guanosine monotonic oligonucleotides capable of adopting a G-quartet structure, are effective inhibitors of aggregation. By utilizing a biochemical/immunoblotting assay as an initial screen, we identified a 20-mer, all G-oligonucleotide (HDG as an active molecule. Subsequent testing in a cell-based assay revealed that HDG was an effective inhibitor of aggregation of a fusion protein, comprised of a mutant Htt fragment and green fluorescent protein (eGFP. Taken together, our results suggest that a monotonic G-oligonucleotide, capable of adopting a G-quartet conformation is an effective inhibitor of aggregation. This oligonucleotide can also enable cell survival in PC12 cells overexpressing a mutant Htt fragment fusion gene. Conclusion Single-stranded DNA oligonucleotides capable of forming stable G-quartets can inhibit aggregation of the mutant Htt fragment protein. This activity maybe an important part of the pathogenecity of Huntington's Disease. Our results reveal a new class of agents that could be developed as a therapeutic approach for Huntington's Disease.

  20. Modulation of 5' splice site selection using tailed oligonucleotides carrying splicing signals

    Directory of Open Access Journals (Sweden)

    Elela Sherif


    Full Text Available Abstract Background We previously described the use of tailed oligonucleotides as a means of reprogramming alternative pre-mRNA splicing in vitro and in vivo. The tailed oligonucleotides that were used interfere with splicing because they contain a portion complementary to sequences immediately upstream of the target 5' splice site combined with a non-hybridizing 5' tail carrying binding sites for the hnRNP A1/A2 proteins. In the present study, we have tested the inhibitory activity of RNA oligonucleotides carrying different tail structures. Results We show that an oligonucleotide with a 5' tail containing the human β-globin branch site sequence inhibits the use of the 5' splice site of Bcl-xL, albeit less efficiently than a tail containing binding sites for the hnRNP A1/A2 proteins. A branch site-containing tail positioned at the 3' end of the oligonucleotide also elicited splicing inhibition but not as efficiently as a 5' tail. The interfering activity of a 3' tail was improved by adding a 5' splice site sequence next to the branch site sequence. A 3' tail carrying a Y-shaped branch structure promoted similar splicing interference. The inclusion of branch site or 5' splice site sequences in the Y-shaped 3' tail further improved splicing inhibition. Conclusion Our in vitro results indicate that a variety of tail architectures can be used to elicit splicing interference at low nanomolar concentrations, thereby broadening the scope and the potential impact of this antisense technology.

  1. Phage annealing proteins promote oligonucleotide-directed mutagenesis in Escherichia coli and mouse ES cells

    Directory of Open Access Journals (Sweden)

    Muyrers Joep PP


    Full Text Available Abstract Background The phage protein pairs, RecE/RecT from Rac or Redα/Redβ from λ, initiate efficient double strand break repair (DSBR in Escherichia coli that has proven very useful for DNA engineering. These phage pairs initiate DSBR either by annealing or by another mechanism that is not defined. Results Here we report that these proteins also mediate single strand oligonucleotide repair (ssOR at high efficiencies. The ssOR activity, unlike DSBR, does not require a phage exonuclease (RecE or Redα but only requires a phage annealing protein (RecT or Redβ. Notably, the P22 phage annealing protein Erf, which does not mediate the same DSBR reactions, also delivers ssOR activity. By altering aspects of the oligonucleotides, we document length and design parameters that affect ssOR efficiency to show a simple relationship to homologies either side of the repair site. Notably, ssOR shows strand bias. Oligonucleotides that can prime lagging strand replication deliver more ssOR than their leading complements. This suggests a model in which the annealing proteins hybridize the oligonucleotides to single stranded regions near the replication fork. We also show that ssOR is a highly efficient way to engineer BACs and can be detected in a eukaryotic cell upon expression of a phage annealing protein. Conclusion Phage annealing proteins can initiate the recombination of single stranded oligonucleotides into endogenous targets in Escherichia coli at very high efficiencies. This expands the repertoire of useful DNA engineering strategies, shows promise for applications in eukaryotic cells, and has implications for the unanswered questions regarding DSBR mediated by RecE/RecT and Redα/Redβ.

  2. Development, characterization and experimental validation of a cultivated sunflower (Helianthus annuus L. gene expression oligonucleotide microarray.

    Directory of Open Access Journals (Sweden)

    Paula Fernandez

    Full Text Available Oligonucleotide-based microarrays with accurate gene coverage represent a key strategy for transcriptional studies in orphan species such as sunflower, H. annuus L., which lacks full genome sequences. The goal of this study was the development and functional annotation of a comprehensive sunflower unigene collection and the design and validation of a custom sunflower oligonucleotide-based microarray. A large scale EST (>130,000 ESTs curation, assembly and sequence annotation was performed using Blast2GO ( The EST assembly comprises 41,013 putative transcripts (12,924 contigs and 28,089 singletons. The resulting Sunflower Unigen Resource (SUR version 1.0 was used to design an oligonucleotide-based Agilent microarray for cultivated sunflower. This microarray includes a total of 42,326 features: 1,417 Agilent controls, 74 control probes for sunflower replicated 10 times (740 controls and 40,169 different non-control probes. Microarray performance was validated using a model experiment examining the induction of senescence by water deficit. Pre-processing and differential expression analysis of Agilent microarrays was performed using the Bioconductor limma package. The analyses based on p-values calculated by eBayes (p<0.01 allowed the detection of 558 differentially expressed genes between water stress and control conditions; from these, ten genes were further validated by qPCR. Over-represented ontologies were identified using FatiScan in the Babelomics suite. This work generated a curated and trustable sunflower unigene collection, and a custom, validated sunflower oligonucleotide-based microarray using Agilent technology. Both the curated unigene collection and the validated oligonucleotide microarray provide key resources for sunflower genome analysis, transcriptional studies, and molecular breeding for crop improvement.

  3. Development, characterization and experimental validation of a cultivated sunflower (Helianthus annuus L.) gene expression oligonucleotide microarray. (United States)

    Fernandez, Paula; Soria, Marcelo; Blesa, David; DiRienzo, Julio; Moschen, Sebastian; Rivarola, Maximo; Clavijo, Bernardo Jose; Gonzalez, Sergio; Peluffo, Lucila; Príncipi, Dario; Dosio, Guillermo; Aguirrezabal, Luis; García-García, Francisco; Conesa, Ana; Hopp, Esteban; Dopazo, Joaquín; Heinz, Ruth Amelia; Paniego, Norma


    Oligonucleotide-based microarrays with accurate gene coverage represent a key strategy for transcriptional studies in orphan species such as sunflower, H. annuus L., which lacks full genome sequences. The goal of this study was the development and functional annotation of a comprehensive sunflower unigene collection and the design and validation of a custom sunflower oligonucleotide-based microarray. A large scale EST (>130,000 ESTs) curation, assembly and sequence annotation was performed using Blast2GO ( The EST assembly comprises 41,013 putative transcripts (12,924 contigs and 28,089 singletons). The resulting Sunflower Unigen Resource (SUR version 1.0) was used to design an oligonucleotide-based Agilent microarray for cultivated sunflower. This microarray includes a total of 42,326 features: 1,417 Agilent controls, 74 control probes for sunflower replicated 10 times (740 controls) and 40,169 different non-control probes. Microarray performance was validated using a model experiment examining the induction of senescence by water deficit. Pre-processing and differential expression analysis of Agilent microarrays was performed using the Bioconductor limma package. The analyses based on p-values calculated by eBayes (p<0.01) allowed the detection of 558 differentially expressed genes between water stress and control conditions; from these, ten genes were further validated by qPCR. Over-represented ontologies were identified using FatiScan in the Babelomics suite. This work generated a curated and trustable sunflower unigene collection, and a custom, validated sunflower oligonucleotide-based microarray using Agilent technology. Both the curated unigene collection and the validated oligonucleotide microarray provide key resources for sunflower genome analysis, transcriptional studies, and molecular breeding for crop improvement.

  4. A study of a steering system algorithm for pleasure boats based on stability analysis of a human-machine system model (United States)

    Ikeda, Fujio; Toyama, Shigehiro; Ishiduki, Souta; Seta, Hiroaki


    Maritime accidents of small ships continue to increase in number. One of the major factors is poor manoeuvrability of the Manual Hydraulic Steering Mechanism (MHSM) in common use. The manoeuvrability can be improved by using the Electronic Control Steering Mechanism (ECSM). This paper conducts stability analyses of a pleasure boat controlled by human models in view of path following on a target course, in order to establish design guidelines for the ECSM. First, to analyse the stability region, the research derives the linear approximated model in a planar global coordinate system. Then, several human models are assumed to develop closed-loop human-machine controlled systems. These human models include basic proportional, derivative, integral and time-delay actions. The stability analysis simulations for those human-machine systems are carried out. The results show that the stability region tends to spread as a ship's velocity increases in the case of the basic proportional human model. The derivative action and time-delay action of human models are effective in spreading the stability region in their respective ranges of frontal gazing points.

  5. Basic Electromagnetism and Materials

    CERN Document Server

    Moliton, André


    Basic Electromagnetism and Materials is the product of many years of teaching basic and applied electromagnetism. This textbook can be used to teach electromagnetism to a wide range of undergraduate science majors in physics, electrical engineering or materials science. However, by making lesser demands on mathematical knowledge than competing texts, and by emphasizing electromagnetic properties of materials and their applications, this textbook is uniquely suited to students of materials science. Many competing texts focus on the study of propagation waves either in the microwave or optical domain, whereas Basic Electromagnetism and Materials covers the entire electromagnetic domain and the physical response of materials to these waves. Professor André Moliton is Director of the Unité de Microélectronique, Optoélectronique et Polymères (Université de Limoges, France), which brings together three groups studying the optoelectronics of molecular and polymer layers, micro-optoelectronic systems for teleco...

  6. Nuclear multifragmentation: Basic concepts

    Indian Academy of Sciences (India)

    G Chaudhuri; S Mallik; S Das Gupta


    We present a brief overview of nuclear multifragmentation reaction. Basic formalism of canonical thermodynamical model based on equilibrium statistical mechanics is described. This model is used to calculate basic observables of nuclear multifragmentation like mass distribution, fragment multiplicity, isotopic distribution and isoscaling. Extension of canonical thermodynamical model to a projectile fragmentation model is outlined. Application of the projectile fragmentation model for calculating average number of intermediate mass fragments and the average size of the largest cluster at different bound, differential charge distribution and cross-section of neutron-rich nuclei of different projectile fragmentation reactions at different energies are described. Application of nuclear multifragmentation reaction in basic research as well as in other domains is outlined.

  7. Binding of Dumbbell Oligonucleotides to MoMuLV Reverse Transcriptase: Inhibitory Properties of RNase H Activity

    Directory of Open Access Journals (Sweden)

    Ajay Kumar


    Full Text Available Dumbbell oligonucleotides with loops of various chemistry were synthesized. Incubation of dumbbell oligonucleotides containing phosphorothioate bonds or trimethylene phosphate linkages in loops with S1 nuclease did not result in significant cleavage under conditions which led to the degradation of dumbbell oligonucleotide containing phophodiester bonds in the loops. The binding of reverse transcriptase of Moloney Murine Leukemia Virus (MoMuLV was evaluated with all the five oligonucleotides. The protein binds to all the dumbbell oligonucleotides with similar affinity. The dissociation constants evaluated using PAGE band mobility shift assays were of the order of 10-7. The inhibitory properties of the retroviral RNase H activity was evaluated using 3H –UTP-labeled RNA:RNA-DNA hybrid. It was found that the best dumbbell oligonucleotide, inhibitor contained phosphorothioate residues in both the loops. Our value studies demonstrated that this particularly designed oligonucleotide displays an IC50 of 18 nM in its inhibition on the reverse transcriptase RNase H activity, a magnitude lower than that of first nucleotide reverse transcriptase of HIV-1, tenofovir, introduced by Gilead Science in the market.

  8. Effect of CD44 Suppression by Antisense Oligonucleotide on Attachment of Human Trabecular Meshwork Cells to HA

    Institute of Scientific and Technical Information of China (English)

    李中国; 张虹


    The effects of suppression of CD44 by CD44-specific antisense oligonucleotide on attachment of human trabecular meshwork cells to hyaluronic acid (HA) were observed and the possible relationship between CD44 and primary open-angle glaucoma (POAG) investigated. CD44-specific antisense oligonucleotide was delivered with cationic lipid to cultured human trabecular meshwork cells. The expression of CD44 suppressed by CD44-specific antisense oligonucleotide was detected by RT-PCR and Western blotting. The effect of CD44 suppression by specific antisense oligonucleotide on attachment of trabecular meshwork cells to HA was measured by MTT assay. Results showed that expression of CD44 was suppressed by CD4, specific antisense oligonucleotide. Antisense oligonucleotide also suppressed the adhesion of human trabecular meshwork cells to HA in a concentration dependent manner. It was concluded that attachment of human trabecular meshwork cells to HA was decreased when CD44 was suppressed by specific antisense oligonucleotide. CD44might play a role in pathogenesis of POAG by affecting the adhesion of trabecular meshwork cells to HA.

  9. Triple helix-forming oligonucleotides conjugated to indolocarbazole poisons direct topoisomerase I-mediated DNA cleavage to a specific site. (United States)

    Arimondo, P B; Bailly, C; Boutorine, A S; Moreau, P; Prudhomme, M; Sun, J S; Garestier, T; Hélène, C


    Topoisomerase I is an ubiquitous DNA-cleaving enzyme and an important therapeutic target in cancer chemotherapy for camptothecins as well as for indolocarbazole antibiotics such as rebeccamycin. To achieve a sequence-specific cleavage of DNA by topoisomerase I, a triple helix-forming oligonucleotide was covalently linked to indolocarbazole-type topoisomerase I poisons. The three indolocarbazole-oligonucleotide conjugates investigated were able to direct topoisomerase I cleavage at a specific site based upon sequence recognition by triplex formation. The efficacy of topoisomerase I-mediated DNA cleavage depends markedly on the intrinsic potency of the drug. We show that DNA cleavage depends also upon the length of the linker arm between the triplex-forming oligonucleotide and the drug. Based on a known structure of the DNA-topoisomerase I complex, a molecular model of the oligonucleotide conjugates bound to the DNA-topoisomerase I complex was elaborated to facilitate the design of a potent topoisomerase I inhibitor-oligonucleotide conjugate with an optimized linker between the two moieties. The resulting oligonucleotide-indolocarbazole conjugate at 10 nM induced cleavage at the triple helix site 2-fold more efficiently than 5 microM of free indolocarbazole, while the other drug-sensitive sites were not cleaved. The rational design of drug-oligonucleotide conjugates carrying a DNA topoisomerase poison may be exploited to improve the efficacy and selectivity of chemotherapeutic cancer treatments by targeting specific genes and reducing drug toxicity.

  10. Decontamination: back to basics. (United States)

    Meredith, Susan J; Sjorgen, Geoff


    My invitation from this Journal's Editor, Felicia Cox, to provide a paper for this themed issue, included the sentence 'I was wondering if you or a colleague would like to contribute a back to basics article on the relevant standards and guidelines for decontamination, including what is compliance?'. The reason it is so interesting to me is that the term 'back to basics' implies reverting to a simpler time in life - when by just sticking to the rules, life became easier. However, with decontamination this is not actually true.

  11. Comprehensive basic mathematics

    CERN Document Server

    Veena, GR


    Salient Features As per II PUC Basic Mathematics syllabus of Karnataka. Provides an introduction to various basic mathematical techniques and the situations where these could be usefully employed. The language is simple and the material is self-explanatory with a large number of illustrations. Assists the reader in gaining proficiency to solve diverse variety of problems. A special capsule containing a gist and list of formulae titled ''REMEMBER! Additional chapterwise arranged question bank and 3 model papers in a separate section---''EXAMINATION CORNER''.

  12. Basic set theory

    CERN Document Server

    Levy, Azriel


    An advanced-level treatment of the basics of set theory, this text offers students a firm foundation, stopping just short of the areas employing model-theoretic methods. Geared toward upper-level undergraduate and graduate students, it consists of two parts: the first covers pure set theory, including the basic motions, order and well-foundedness, cardinal numbers, the ordinals, and the axiom of choice and some of it consequences; the second deals with applications and advanced topics such as point set topology, real spaces, Boolean algebras, and infinite combinatorics and large cardinals. An

  13. Basic research championed (United States)

    Friebele, Elaine

    In April, the Office of National Science and Technology Policy released its biennial report to Congress. Science and Technology: Shaping the Twenty-First Century addresses the President's policy for maintaining U.S. leadership in science and technology, significant developments, and important national issues in science, and opportunities to use science and technology in federal programs and national goals. The administration strongly supports basic research as a sound investment and an inspiration to society. As corporate laboratories increasingly favor applied R&D projects, the federal government is becoming the dominant sponsor of long-term, basic research.

  14. Basic properties of semiconductors

    CERN Document Server

    Landsberg, PT


    Since Volume 1 was published in 1982, the centres of interest in the basic physics of semiconductors have shifted. Volume 1 was called Band Theory and Transport Properties in the first edition, but the subject has broadened to such an extent that Basic Properties is now a more suitable title. Seven chapters have been rewritten by the original authors. However, twelve chapters are essentially new, with the bulk of this work being devoted to important current topics which give this volume an almost encyclopaedic form. The first three chapters discuss various aspects of modern band theory and the

  15. Basic Financial Accounting

    DEFF Research Database (Denmark)

    Wiborg, Karsten

    This textbook on Basic Financial Accounting is targeted students in the economics studies at universities and business colleges having an introductory subject in the external dimension of the company's economic reporting, including bookkeeping, etc. The book includes the following subjects: busin......: business entities, the transformation process, types of businesses, stakeholders, legislation, the annual report, the VAT system, double-entry bookkeeping, inventories, and year-end cast flow analysis.......This textbook on Basic Financial Accounting is targeted students in the economics studies at universities and business colleges having an introductory subject in the external dimension of the company's economic reporting, including bookkeeping, etc. The book includes the following subjects...

  16. Commuter exposure to black carbon, carbon monoxide, and noise in the mass transport khlong boats of Bangkok, Thailand (United States)

    Ziegler, A. D.; Velasco, E.; Ho, K. J.


    Khlong (canal) boats are a unique mass transport alternative in the congested city of Bangkok. Canals and rivers provide exclusive transit-ways for reducing the commuting time of thousands of city residents daily. However, as a consequence of the service characteristics and boats design and state of repair, they can represent a potential public health risk and an important source of black carbon and greenhouse gases. This work quantifies commuter exposure to black carbon, CO and noise when waiting for and travelling in these diesel fueled boats. Exposure to toxic pollutants and acute noise is similar or worse than for other transportation modes. Mean black carbon concentrations observed at one busy pier and along the main canal were much higher than ambient concentrations at sites impacted by vehicular traffic. Concentrations of CO were similar to those reported for roadside areas of Bangkok. The equivalent continuous sound levels registered at the landing pier were similar to those reported for roadsides, but values recorded inside the boats were significantly higher. We believe that the boat service is a viable alternative mode of mass transport, but public safety could be improved to provide a high quality service, comparable to modern rail systems or emerging bus rapid transit systems. These investments would also contribute to reduce the emission of black carbon and other greenhouse and toxic pollutants.

  17. Modeling the impacts of cetacean-focused tourism in Taiwan: observations from cetacean watching boats: 2002-2005. (United States)

    Tseng, Yung-Ping; Huang, Yu-Chin; Kyle, Gerard T; Yang, Ming-Ching


    Cetacean-focused tourism in Taiwan has grown rapidly since 1997. This development, measured in terms of both number of tour boats and visitors, has resulted in many resource management challenges stemming from the absence of regulation and scientific data. To fill this void in empirical evidence, we used 464 sighting records from 2002 to 2005 to model the impact of cetacean-focused tourism. Cox proportional hazard analysis indicated cetacean avoidance responses to cetacean watching boats were strongly associated with pod size, mother-calf pairs, and cetacean-vessel distances. Mother-calf pairs abandoned their avoidance tactic by 55% compared to noncalf groups when tour boats approached. Second, the hazard ratio of abundance was 0.996, suggesting that the odds of encountering avoidance responses by the cetaceans decreased by 42% for every 100-member increase in the cetacean pod size. Last, distances maintained by boats from the cetaceans was positively related to avoidance responses (i.e., less avoidance behavior with closer interaction). Based on our findings, we have the following recommendations: (a) limit vessels from approaching mothers with calves, (b) limit vessels from approaching small groups of cetaceans, (c) reduced avoidance behavior to boat traffic may be a red flag for potential long-term disturbance, and (d) apply the "precautionary principle" based on the best scientific information available in cetacean-based tourism in Taiwan. These recommendations will help contribute to the sustainable development of cetacean-focused tourism in Taiwan.

  18. Quantification of Boat Visitation Rates at Artificial and Natural Reefs in the Eastern Gulf of Mexico Using Acoustic Recorders. (United States)

    Simard, Peter; Wall, Kara R; Mann, David A; Wall, Carrie C; Stallings, Christopher D


    Artificial reefs are commonly used as a management tool, in part to provide ecosystem services, including opportunities for recreational fishing and diving. Quantifying the use of artificial reefs by recreational boaters is essential for determining their value as ecosystem services. In this study, four artificial-natural reef pairs in the eastern Gulf of Mexico (off western Florida) were investigated for boat visitation rates using autonomous acoustic recorders. Digital SpectroGram (DSG) recorders were used to collect sound files from April 2013 to March 2015. An automatic detection algorithm was used to identify boat noise in individual files using the harmonic peaks generated by boat engines, and by comparing the sound amplitude of each file with surrounding files. In all four pairs, visitation rates were significantly higher at the artificial reef than the natural reef. This increase in boat visitation was likely due to actual or perceived increased quality of fishing and diving at the artificial reefs, or to lack of knowledge of the presence or locations of the natural reefs. Inshore reefs (25 m depth). This study appears to be the first to use acoustic data to measure participant use of boating destinations, and highlights the utility of acoustic monitoring for the valuation of this important ecosystem service provided by artificial reefs.

  19. Quantification of Boat Visitation Rates at Artificial and Natural Reefs in the Eastern Gulf of Mexico Using Acoustic Recorders (United States)

    Simard, Peter; Wall, Kara R.; Mann, David A.; Wall, Carrie C.; Stallings, Christopher D.


    Artificial reefs are commonly used as a management tool, in part to provide ecosystem services, including opportunities for recreational fishing and diving. Quantifying the use of artificial reefs by recreational boaters is essential for determining their value as ecosystem services. In this study, four artificial–natural reef pairs in the eastern Gulf of Mexico (off western Florida) were investigated for boat visitation rates using autonomous acoustic recorders. Digital SpectroGram (DSG) recorders were used to collect sound files from April 2013 to March 2015. An automatic detection algorithm was used to identify boat noise in individual files using the harmonic peaks generated by boat engines, and by comparing the sound amplitude of each file with surrounding files. In all four pairs, visitation rates were significantly higher at the artificial reef than the natural reef. This increase in boat visitation was likely due to actual or perceived increased quality of fishing and diving at the artificial reefs, or to lack of knowledge of the presence or locations of the natural reefs. Inshore reefs (25 m depth). This study appears to be the first to use acoustic data to measure participant use of boating destinations, and highlights the utility of acoustic monitoring for the valuation of this important ecosystem service provided by artificial reefs. PMID:27500533

  20. Modeling the Impacts of Cetacean-Focused Tourism in Taiwan: Observations from Cetacean Watching Boats: 2002-2005 (United States)

    Tseng, Yung-Ping; Huang, Yu-Chin; Kyle, Gerard T.; Yang, Ming-Ching


    Cetacean-focused tourism in Taiwan has grown rapidly since 1997. This development, measured in terms of both number of tour boats and visitors, has resulted in many resource management challenges stemming from the absence of regulation and scientific data. To fill this void in empirical evidence, we used 464 sighting records from 2002 to 2005 to model the impact of cetacean-focused tourism. Cox proportional hazard analysis indicated cetacean avoidance responses to cetacean watching boats were strongly associated with pod size, mother-calf pairs, and cetacean-vessel distances. Mother-calf pairs abandoned their avoidance tactic by 55% compared to noncalf groups when tour boats approached. Second, the hazard ratio of abundance was 0.996, suggesting that the odds of encountering avoidance responses by the cetaceans decreased by 42% for every 100-member increase in the cetacean pod size. Last, distances maintained by boats from the cetaceans was positively related to avoidance responses (i.e., less avoidance behavior with closer interaction). Based on our findings, we have the following recommendations: (a) limit vessels from approaching mothers with calves, (b) limit vessels from approaching small groups of cetaceans, (c) reduced avoidance behavior to boat traffic may be a red flag for potential long-term disturbance, and (d) apply the "precautionary principle" based on the best scientific information available in cetacean-based tourism in Taiwan. These recommendations will help contribute to the sustainable development of cetacean-focused tourism in Taiwan.

  1. OligoPrep PVA support for oligonucleotide synthesis in columns on a scale up to 10 micromol. (United States)

    Aitken, Sheena; Anderson, Emma


    OligoPrep is a macroporous polyvinylacetate (PVA) biodegradable support that has been designed for cost-effective automated synthesis of oligonucleotides using standard phosphoramidite chemistry. Originally developed for large-scale oligonucleotide synthesis in beds and reactors, we present here its utility for medium-scale work of 1-10 micromol in column syntheses on standard DNA synthesizers. We show how an increase in scale, and, therefore, yield, can be achieved without significant increase in reagent quantity. Additional deblock and oxidation cycles can provide high coupling yields, and the use of concentrated ammonia in aqueous methylamine (AMA) for oligonucleotide cleavage and deprotection results in excellent recovery.

  2. Amino acids attached to 2'-amino-LNA: Synthesis of DNA mixmer oligonucleotides with increased duplex stability

    DEFF Research Database (Denmark)

    Johannsen, Marie Willaing; Wengel, Jesper; Wamberg, Michael Chr.;


    The synthesis of 2'-amino-LNA (locked nucleic acid) opens up exciting possibilities for modification of nucleic acids by conjugation to the 2'-nitrogen. Incorporation of unmodified and N-functionalized 2'-amino-LNA nucleotides improve duplex stability compared to unmodified DNA. 2'-Amino......-LNA nucleosides derivatized with amino acids have been synthesized and incorporated into DNA oligonucleotides. Following oligonucleotide synthesis, peptides have been added using solid phase peptide coupling chem. Modification of oligonucleotides with pos. charged residues greatly improves thermal stability....

  3. Optimization and scale-up of oligonucleotide synthesis in packed bed reactors using computational fluid dynamics modeling. (United States)

    Wolfrum, Christian; Josten, Andre; Götz, Peter


    A computational fluid dynamics (CFD) model for the analysis of oligonucleotide synthesis in packed bed reactors was developed and used to optimize the scale up of the process. The model includes reaction kinetics data obtained under well defined conditions comparable to the situation in the packed bed. The model was validated in terms of flow conditions and reaction kinetics by comparison with experimental data. Experimental validation and the following model parameter studies by simulation were performed on the basis of a column with 0.3 g oligonucleotide capacity. The scale-up studies based on CFD modelling were calculated on a 440 g scale (oligonucleotide capacity).

  4. Characterization of the nanostructure of complexes formed by single- or double-stranded oligonucleotides with a cationic surfactant. (United States)

    Liu, Xiaoyang; Abbott, Nicholas L


    We report the use of dynamic light scattering (DLS), small-angle neutron scattering (SANS), and small-angle X-ray scattering (SAXS) to characterize the nanostructure of complexes formed by either single- or double-stranded oligonucleotides with a cationic surfactant (cetyltrimethylammonium bromide, CTAB) in aqueous solution (1 mM Li(2)SO(4)). For single-stranded oligonucleotides 5'-A(20)-3' and 5'-CCCCATTCTAGCAGCCCGGG-3', both the appearance of two Bragg peaks (at 0.14 and 0.28 Å(-1)) in SAXS spectra with a spacing of 1:2 and form factor fits to SANS spectra are consistent with the presence of multilamellar vesicles (with, on average, 6-9 layers with a periodicity of 45-48 Å). Some samples showed evidence of an additional Bragg peak (at 0.20 Å(-1)) associated with periodic packing (with a periodicity of 31 Å) of the oligonucleotides within the lamellae of the nanostructure. The nucleotide composition of the single-stranded oligonucleotides was also found to impact the number and size of the complexes formed with CTAB. In contrast to 5'-A(20)-3' and 5'-CCCCATTCTAGCAGCCCGGG-3', 5'-T(20)-3' did not change the state of aggregation of CTAB (globular micelles) over a wide range of oligonucleotide:CTAB charge ratios. These results support the proposition that hydrophobic interactions, as well as electrostatics, play a central role in the formation of complexes between cationic amphiphiles and single-stranded oligonucleotides and thus give rise to nanostructures that depend on nucleotide composition. In contrast to the single-stranded oligonucleotides, for double-stranded oligonucleotides mixed with CTAB, three Bragg peaks (0.13, 0.23, and 0.25 Å(-1)) in SAXS spectra with a spacing ratio of 1:√3:√4 and characteristic changes in SANS spectra indicate formation of a hexagonal nanostructure. Also, the composition of the double-stranded oligonucleotides did not measurably impact the nanostructure of complexes formed with CTAB, suggesting that electrostatic

  5. Ethanol Basics (Fact Sheet)

    Energy Technology Data Exchange (ETDEWEB)


    Ethanol is a widely-used, domestically-produced renewable fuel made from corn and other plant materials. More than 96% of gasoline sold in the United States contains ethanol. Learn more about this alternative fuel in the Ethanol Basics Fact Sheet, produced by the U.S. Department of Energy's Clean Cities program.

  6. Korean Basic Course. (United States)

    Defense Language Inst., Washington, DC.

    These 11 volumes of the Korean Basic Course comprise 112 lesson units designed to train native English language speakers to Level 3 proficiency in comprehension and speaking and Level 2 proficiency in reading and writing Korean. (Level 5 on this scale is native-speaker level.) Intended for classroom use in the Defense Language Institute intensive…

  7. Basic physics for all

    CERN Document Server

    Kumar, B N


    This is a simple, concise book for both student and non-physics students, presenting basic facts in straightforward form and conveying fundamental principles and theories of physics. This book will be helpful as a supplement to class teaching and to aid those who have difficulty in mastering concepts and principles.

  8. Vaccine Basics (Smallpox) (United States)

    ... this page: About . Smallpox About Smallpox History of Smallpox Spread and Eradication of Smallpox Transmission Signs and Symptoms Prevention and Treatment Smallpox Vaccine Basics Vaccine Safety Side Effects of Vaccination Who Should Get a Smallpox Vaccination? Bioterrorism The ...

  9. FULA BASIC COURSE. (United States)



  10. Basic Library List. (United States)

    Duren, William L., Jr.

    Reported is an initial attempt to define a minimal college mathematics library. Included is a list of some 300 books, from which approximately 170 are to be chosen to form a basic library in undergraduate mathematics. The areas provided for in this list include Algebra, Analysis, Applied Mathematics, Geometry, Topology, Logic, Foundations and Set…

  11. Lippincott Basic Reading Program. (United States)

    Monterey Peninsula Unified School District, Monterey, CA.

    This program, included in "Effective Reading Programs...," serves 459 students in grades 1-3 at 15 elementary schools. The program employs a diagnostic-prescriptive approach to instruction in a nongraded setting through the use of the Lippincott Basic Reading program. When a child enters the program, he is introduced to a decoding…

  12. Basic Drafting: Book Two. (United States)

    Davis, Ronald; And Others

    The second of a two-book course in drafting, this manual consists of 12 topics in the following units: sketching techniques, geometric constructions, orthographic views, dimensioning procedures, basic tolerancing, auxiliary views, sectional views, inking tools and techniques, axonometrics, oblique, perspective, and computer-aided drafting.…

  13. Health Insurance Basics (United States)

    ... members at a lower cost. The four basic types of managed care plans are: HMO (Health Maintenance Organization). When you join an HMO, you choose a ... may have to pay more. EPO (Exclusive Provider Organization). An EPO is like a PPO, only ... Health Plan (CDHP) This type of plan is fairly new. It lets you ...

  14. Basic bioreactor design.

    NARCIS (Netherlands)

    Riet, van 't K.; Tramper, J.


    Based on a graduate course in biochemical engineering, provides the basic knowledge needed for the efficient design of bioreactors and the relevant principles and data for practical process engineering, with an emphasis on enzyme reactors and aerated reactors for microorganisms. Includes exercises.

  15. Basic Nuclear Physics. (United States)

    Bureau of Naval Personnel, Washington, DC.

    Basic concepts of nuclear structures, radiation, nuclear reactions, and health physics are presented in this text, prepared for naval officers. Applications to the area of nuclear power are described in connection with pressurized water reactors, experimental boiling water reactors, homogeneous reactor experiments, and experimental breeder…

  16. Canadian Adult Basic Education. (United States)

    Brooke, W. Michael, Comp.

    "Trends," a publication of the Canadian Association for Adult Education, is a collection of abstracts on selected subjects affecting adult education; this issue is on adult basic education (ABE). It covers teachers and teacher training, psychological factors relating to the ABE teacher and students, manuals for teachers, instructional…

  17. Basic Microfluidics Theory

    DEFF Research Database (Denmark)

    Svendsen, Winnie Edith


    ,000 m−1, which is a huge difference and has a large impact on flow behavior. In this chapter the basic microfluidic theory will be presented, enabling the reader to gain a comprehensive understanding of how liquids behave at the microscale, enough to be able to engage in design of micro systems...

  18. Basic Tuberculosis Facts

    Centers for Disease Control (CDC) Podcasts


    In this podcast, Dr. Kenneth Castro, Director of the Division of Tuberculosis Elimination, discusses basic TB prevention, testing, and treatment information.  Created: 3/12/2012 by National Center for HIV/AIDS, Viral Hepatitis, STD, and TB Prevention (NCHHSTP).   Date Released: 3/12/2012.

  19. The Boat-coffins and Single-log Coffins in Shangye Street, Chengdu, Sichuan

    Institute of Scientific and Technical Information of China (English)

    蒋成; 颜劲松; 陈云洪; 刘雨茂; 王淑芳


    The boat-coffins and single-log coffins are located in No. 58, Shangye 商业 Street, within the first ring road of Chengdu 成都 downtown area. From July 2000 to the beginning of 2001 Chengdu Municipal Institute of Antiquity and Archaeology made an excavation of this site and determined it to be a massive tomb of the early Warring States period. It was selected as one of the top ten great archaeological discoveries in China in 2000 and declared by the State Council as a historical site of the fifth group under special protection within China in July 2001.

  20. Recreational Boat Safety Collision Research - Phase 2. Volume 2. Collision Accident Investigations - 1975 (United States)


    greater on every weekday except Wednesday than on Saturday or Sunday. Perhaps boaters are more careful on weekends when waterways are more crowded...that his boat was for rer, so more care could have been given to lookout for the water scooter. Use of alcohol bf no. 4 may have caused him not to...batterry and fuell tank al h if itulewae udrsa 90 DAY ~1 / 0% Mane plywood * Telescopc peddle 0 Self beling GENTLE AS A KITTEN , due to Hydro-Cycle’s unique

  1. A study of the sewn-plank boats of Goa, India

    Digital Repository Service at National Institute of Oceanography (India)

    Shaikh, Z.A; Tripati, S.; Shinde, V.

    from local saw mills or brought from neighbouring states. After cutting and trimming the timber in the saw mills, they are brought to the yard. The construction of a sewn plank built boat starts by laying the keel (Fig. 5). The keel defines the size... stream_size 26146 stream_content_type text/plain stream_name Int_J_Naut_Archaeol_41_148a.pdf.txt stream_source_info Int_J_Naut_Archaeol_41_148a.pdf.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 1...

  2. Study Project Comprising Two Surveys on Behalf of the National Association of State Boating Law Administrators. (United States)


    units been involved in a search for persons thought to be lost in the water? Boat Swimmer Hunter-Fisherman Aircraft Total 11 Would staff of your SAR...Governors Island. Our search plan coincides with standard procedures (i.e.) expanded search, etc. 5. Occurrences involving swimmers and surfers have...0 :D 4Jf 10 OU 4 PC to 41N~ 0~ 0 lue ~ --4 0 0 m4O0 ~ 4~ Z) ts Z 0 - - imt < U- 4 ") 4H 0 u0 u i 69 CD 00 o 0000 0 0 0 S0cJ N Oa O 0 %D %.0~o o..-r O

  3. Weekday/weekend differences in gasoline related hydrocarbons at coastal PAMS sites due to recreational boating (United States)

    Henry, Robert F.


    Analysis of PAMS (Photochemical Assessment Monitoring Stations) data at several coastal sites reveals large weekday/weekend differences in gasoline related hydrocarbons. Elevated concentrations of gasoline related constituents, including alkanes, alkenes, and aromatics, are observed on weekends at the PAMS monitors at Sherwood Island State Park in Westport, CT and at Newbury, MA. An analysis of the ratio of the concentrations of 2,3-dimethylbutane to 2,2-dimethylbutane indicates these compounds are freshly emitted, and an investigation in conjunction with wind data shows that the elevated concentrations are associated primarily with onshore winds. These elevated concentrations are most likely due to weekend recreational boating.

  4. Developing the Navy’s NC Flying Boats: Transforming Aeronautical Engineering for the First Transatlantic Flight (United States)


    ft. Max Gross Weight 28,0001b Cruising Speed 84kts Range 1,278 nm Crew 6 Engines (4) 400 hp Liberty L-12 Figure 7. The NC flying boat. In...of the hull. The keel of the hull is built up from spruce while the bottom planking is laid up from two plies of cedar separated by a waterproofing...pilots could remain below decks and out of the weather if desired. The aircraft commander even had enough space to lie down on the planking that made up

  5. Influence of boat noises on escape behaviour of white-spotted eagle ray Aetobatus ocellatus at Moorea Island (French Polynesia). (United States)

    Berthe, Cecile; Lecchini, David


    The present study tested different sounds that could disturb eagle rays (Aetobatus ocellatus) during their foraging activities at Moorea, French Polynesia. Results showed that artificial white sound and single-frequency tones (40 Hz, 600 Hz or 1 kHz) did not have an effect on rays (at least 90% of rays continued to forage over sand), while playbacks of boat motor sound significantly disturbed rays during foraging activity (60% exhibited an escape behaviour). Overall, our study highlighted the negative effect of boat noises on the foraging activity of eagle rays. These noises produced by boat traffic could, however, have some positive effects for marine aquaculture if they could be used as a deterrent to repel the eagle rays, main predators of the pearl oysters.

  6. Internal conflicts of Emily Dickinson-A study of the images of boat and sea in Dickinson’s poems

    Institute of Scientific and Technical Information of China (English)

    WAN Hai-yan


    The peculiar images, the boat and the sea, often recur in the poems of Emily Dickinson, the great American poet, which well reflect her internal conflicts, the conflicts between being an autonomous woman and being a passionate yet submissive wom-an. In her poems, the sea is a symbol of the male-dominated society or an individual man. While the little boat is the epitome of Dickinson:sometimes it wants to be strong-willed and clear of its destination, sometimes it just wants to moor in the sea and en-joy the calmness. The paper tends to reveal the psychological struggle of Emily Dickinson and show the whole image of her as an individual through the analysis of the images of boat and sea in Dickinson’s poems.

  7. Development of boats propelled by fuel cells; Developpement d'embarcations mues par piles a combustible

    Energy Technology Data Exchange (ETDEWEB)

    Affolter, J.F.; Mariotti, P. [Ecole d' Ingenieurs du Canton de Vaud, HES-SO, Yverdon les Bains (Switzerland)


    An important European demand for propulsion systems lead us to think that the number of electric ships is growing rapidly. A lot of lakes in Europe are not allowed to the thermal navigation. The aim is to offer a maximum of comfort, reliability, silence, lack of pollution and respect to environment. One of the answers to this expectation is the use of the electric propulsion and a generation of power with fuel cells. After having fabricated the 'Hydroxy100', a single-seater boat propelled by a 100 W fuel cell, and a two-seater boat designed for a 300 W fuel cell, a new plan for a six-seater pleasure boat is being studied. These pilot plans are carried out with the aim to favour the development of this technology. (O.M.)

  8. Modified technique of using conventional slider boat for liquid phase epitaxy of silicon for solar cell application

    Indian Academy of Sciences (India)

    D Majumdar; S Chatterjee; U Gangopadhyay; H Saha


    Epitaxial layers of silicon are grown on single crystal Si-substrate from a solution of silicon in indium using conventional graphite slider boat technique. The important problems of natural convection due to lower density of silicon compared to indium, poor wetting of substrate due to high angle of contact of indium solution on silicon substrate resulting in poor nucleation, melt removal from the growth substrate and saturation wafer associated with LPE in this technique are practically eliminated using sandwich method with simple modifications of the boat and the method of growth. Some experimental studies on the effect of different surface preparations of growth substrate are also reported. Growth results are shown and discussed. Further, improvization of slider boat to facilitate better study of growth parameters is suggested in the line of modification already carried out.

  9. 经编无缝船袜的开发%Development of Warp Knitted Seamless Boat Socks

    Institute of Scientific and Technical Information of China (English)

    姚馨馨; 丛洪莲


    经编无缝船袜是由双针床拉舍尔经编机生产的一次成型织物。船袜又称“隐形袜”,鞋子可将袜面部分罩住。经编无缝船袜款式多样,花型丰富,且无缝合,吸湿排汗,穿着十分舒适。本课题介绍了船袜的生产设备、多款经编无缝船袜的不同特点,选用花型时的注意点以及船袜的功能性特点,如利用硅胶使得经编无缝船袜摒弃易滑脱的缺陷等。简单介绍船袜的开发过程。未来经编无缝船袜需要从新原料、新款式、新工艺等角度出发,获得更广阔的市场。%Warp knitted seamless boat socks were produced by a double bar raschel machine. It is called in-visible boat socks whose face can be covered by shoes. Warp knitted seamless boat socks have diverse styles and rich patterns, they are moisture absorbable, breathable and comfortable. Some main points of boat socks were in-troduced such as the production equipment, different characteristics with diverse styles, key points for selecting patterns and its functional features such as the use of silica gel which made the warp knitted seamless boat socks hard to slip. The development process of boat socks was briefly introduced. From the perspective of new materi-als, new style and new technology, the future of warp knitted seamless socks will have a broader market.

  10. Synthesis of the Tellurium-Derivatized Phosphoramidites and their Incorporation into DNA Oligonucleotides (United States)

    Jiang, Sibo; Sheng, Jia


    Introduction In this unit, an efficient method for the synthesis of 2’-tellerium modified phosphoramidite and its incorporation into oligonucleotide are presented. We choose 5’-O-DMTr-2,2’-anhydro-uridine and -thymidine nucleosides (S.1, S.2) as starting materials due to their easy preparation. The 5’-O-DMTr-2,2’-anhydro-uridine and -thymidine can be converted to corresponding the 2’-tellerium-derivatized nucleosides by treating with the telluride nucleophiles. Subsequently, the 2’-Te-nucleosides can be transformed into 3’-phosphoramidites, which are the building blocks for DNA/RNA synthesis. The DNA synthesis, purification and applications of oligonucleotides containing 2’-Te-U or 2’-Te-T are described in this protocol. PMID:22147418

  11. Ultramild protein-mediated click chemistry creates efficient oligonucleotide probes for targeting and detecting nucleic acids

    DEFF Research Database (Denmark)

    Nåbo, Lina J.; Madsen, Charlotte Stahl; Jensen, Knud Jørgen


    results by electronic structure calculations. Functionalized oligonucleotides were prepared in good yields by protein-mediated CuAAC click reactions for the first time with a human copper-binding chaperon. The carbohydrate, peptide, and fluorescent derivatives display high binding affinity and selectivity...... targeting and detection properties. We focus in particular on the pH sensitivity of these new probes and their high target specificity. For the first time, human copper(I)-binding chaperon Cox17 was applied to effectively catalyze click labeling of oligonucleotides. This was performed under ultramild...... conditions with fluorophore, peptide, and carbohydrate azide derivatives. In thermal denaturation studies, the modified probes showed specific binding to complementary DNA and RNA targets. Finally, we demonstrated the pH sensitivity of the new rhodamine-based fluorescent probes in vitro and rationalize our...

  12. Clinical potential of oligonucleotide-based therapeutics in the respiratory system. (United States)

    Moschos, Sterghios A; Usher, Louise; Lindsay, Mark A


    The discovery of an ever-expanding plethora of coding and non-coding RNAs with nodal and causal roles in the regulation of lung physiology and disease is reinvigorating interest in the clinical utility of the oligonucleotide therapeutic class. This is strongly supported through recent advances in nucleic acids chemistry, synthetic oligonucleotide delivery and viral gene therapy that have succeeded in bringing to market at least three nucleic acid-based drugs. As a consequence, multiple new candidates such as RNA interference modulators, antisense, and splice switching compounds are now progressing through clinical evaluation. Here, manipulation of RNA for the treatment of lung disease is explored, with emphasis on robust pharmacological evidence aligned to the five pillars of drug development: exposure to the appropriate tissue, binding to the desired molecular target, evidence of the expected mode of action, activity in the relevant patient population and commercially viable value proposition.

  13. The Role of Fluorinated Alcohols as Mobile Phase Modifiers for LC-MS Analysis of Oligonucleotides (United States)

    Basiri, Babak; van Hattum, Hilde; van Dongen, William D.; Murph, Mandi M.; Bartlett, Michael G.


    Hexafluoroisopropanol (HFIP) has been widely used as an acidic modifier for mobile phases for liquid chromatography-mass spectrometry (LC-MS) analysis of oligonucleotides ever since the first report of its use for this purpose. This is not surprising, considering the exceptional performance of HFIP compared with carboxylic acids, which cause significant MS signal suppression in electrospray ionization. However, we have found that other fluorinated alcohols can also be utilized for mobile phase preparation and the choice of optimal fluorinated alcohol is determined by the ion-pairing (IP) agent. Although HFIP is a very good choice to be used alongside less hydrophobic IP agents, other fluorinated alcohols such as 1,1,1,3,3,3-hexafluoro-2-methyl-2-propanol (HFMIP) can significantly outperform HFIP when used with more hydrophobic IP agents. We also found that more acidic fluorinated alcohols assist with the transfer of oligonucleotides with secondary structure (e.g., folded strands and hairpins) into the gas phase.

  14. Heated oligonucleotide ligation assay (HOLA): an affordable single nucleotide polymorphism assay. (United States)

    Black, W C; Gorrochotegui-Escalante, N; Duteau, N M


    Most single nucleotide polymorphism (SNP) detection requires expensive equipment and reagents. The oligonucleotide ligation assay (OLA) is an inexpensive SNP assay that detects ligation between a biotinylated "allele-specific detector" and a 3' fluorescein-labeled "reporter" oligonucleotide. No ligation occurs unless the 3' detector nucleotide is complementary to the SNP nucleotide. The original OLA used chemical denaturation and neutralization. Heated OLA (HOLA) instead uses a thermal stable ligase and cycles of denaturing and hybridization for ligation and SNP detection. The cost per genotype is approximately US$1.25 with two-allele SNPs or approximately US$1.75 with three-allele SNPs. We illustrate the development of HOLA for SNP detection in the Early Trypsin and Abundant Trypsin loci in the mosquito Aedes aegypti (L.) and at the a-glycerophosphate dehydrogenase locus in the mosquito Anopheles gambiae s.s.

  15. Ultrahigh molecular recognition specificity of competing DNA oligonucleotide strands in thermal equilibrium

    CERN Document Server

    Schenkelberger, Marc; Mai, Timo; Ott, Albrecht


    The specificity of molecular recognition is important to molecular self-organization. A prominent example is the biological cell where, within a highly crowded molecular environment, a myriad of different molecular receptor pairs recognize their binding partner with astonishing accuracy. In thermal equilibrium it is usually admitted that the affinity of recognizer pairs only depends on the nature of the two binding molecules. Accordingly, Boltzmann factors of binding energy differences relate the molecular affinities among different target molecules that compete for the same probe. Here, we consider the molecular recognition of short DNA oligonucleotide single strands. We show that a better matching oligonucleotide strand can prevail against a disproportionally more concentrated competitor that exhibits reduced affinity due to a mismatch. The magnitude of deviation from the simple picture above may reach several orders of magnitude. In our experiments the effective molecular affinity of a given strand remains...

  16. Computer selection of oligonucleotide probes from amino acid sequences for use in gene library screening. (United States)

    Yang, J H; Ye, J H; Wallace, D C


    We present a computer program, FINPROBE, which utilizes known amino acid sequence data to deduce minimum redundancy oligonucleotide probes for use in screening cDNA or genomic libraries or in primer extension. The user enters the amino acid sequence of interest, the desired probe length, the number of probes sought, and the constraints on oligonucleotide synthesis. The computer generates a table of possible probes listed in increasing order of redundancy and provides the location of each probe in the protein and mRNA coding sequence. Activation of a next function provides the amino acid and mRNA sequences of each probe of interest as well as the complementary sequence and the minimum dissociation temperature of the probe. A final routine prints out the amino acid sequence of the protein in parallel with the mRNA sequence listing all possible codons for each amino acid.

  17. Photoswitchable oligonucleotide-modified gold nanoparticles: controlling hybridization stringency with photon dose. (United States)

    Yan, Yunqi; Chen, Jennifer I L; Ginger, David S


    We describe a new class of stimulus-responsive DNA-functionalized gold nanoparticles that incorporate azobenzene-modified oligonucleotides. Beyond the classic directed assembly and sensing behaviors associated with oligonucleotide-modified nanoparticles, these particles also exhibit reversible photoswitching of their assembly behavior. Exposure to UV light induces a trans-cis isomerization of the azobenzene which destabilizes the DNA duplex, resulting in dissociation of the nanoparticle assemblies. The isomerization is reversible upon exposure to blue light, resulting in rehybridization and reassembly of the DNA-linked nanoparticle clusters. We show that perfectly complementary and partially mismatched strands exhibit clearly distinguishable photoinduced melting properties, and we demonstrate that photon dose can thus be used in place of temperature or ionic strength to control hybridization stringency with the ability to discriminate single-base mismatches.

  18. Detection and identification of intestinal pathogenic bacteria by hybridization to oligonucleotide microarrays

    Institute of Scientific and Technical Information of China (English)

    Lian-Qun Jin; Jun-Wen Li; Sheng-Qi Wang; Fu-Huan Chao; Xin-Wei Wang; Zheng-Quan Yuan


    AIM: To detect the common intestinal pathogenic bacteria quickly and accurately.METHODS: A rapid (<3 h) experimental procedure was set up based upon the gene chip technology. Target genes were amplified and hybridized by oligonucleotide microarrays.RESULTS: One hundred and seventy strains of bacteria in pure culture belonging to 11 genera were successfully discriminated under comparatively same conditions, and a series of specific hybridization maps corresponding to each kind of bacteria were obtained. When this method was applied to 26 divided cultures, 25 (96.2%) were identified.CONCLUSION: Salmonella sp., Escherichia coli, Shigella sp., Listeria monocytogenes, Vibrio parahaemolyticus,Staphylococcus aureus, Proteus sp., Bacillus cereus,Vibrio cholerae, Enterococcus faecalis, Yersinia enterocolitica, and Campylobacter jejuni can be detected and identified by our microarrays. The accuracy, range,and discrimination power of this assay can be continually improved by adding further oligonucleotides to the arrays without any significant increase of complexity or cost.

  19. Antibacterial Activity of DNA-Stabilized Silver Nanoclusters Tuned by Oligonucleotide Sequence. (United States)

    Javani, Siamak; Lorca, Romina; Latorre, Alfonso; Flors, Cristina; Cortajarena, Aitziber L; Somoza, Álvaro


    Silver nanoclusters (AgNCs) stabilized by DNA are promising materials with tunable fluorescent properties, which have been employed in a plethora of sensing systems. In this report, we explore their antimicrobial properties in Gram-positive and Gram-negative bacteria. After testing 9 oligonucleotides with different sequence and length, we found that the antibacterial activity depends on the sequence of the oligonucleotide employed. The sequences tested yielded fluorescent AgNCs, which can be grouped in blue, yellow, and red emitters. Interestingly, blue emitters yielded poor antibacterial activity, whereas yellow and red emitters afforded an activity similar to silver nitrate. Furthermore, structural studies using circular dichroism indicate the formation of complexes with different stability and structure, which might be one of the factors that modulate their activity. Finally, we prepared a trimeric structure containing the sequence that afforded the best antimicrobial activity, which inhibited the growth of Gram-positive and negative bacteria in the submicromolar range.

  20. Efficient Synthesis and Biological Evaluation of 5'-GalNAc Conjugated Antisense Oligonucleotides. (United States)

    Østergaard, Michael E; Yu, Jinghua; Kinberger, Garth A; Wan, W Brad; Migawa, Michael T; Vasquez, Guillermo; Schmidt, Karsten; Gaus, Hans J; Murray, Heather M; Low, Audrey; Swayze, Eric E; Prakash, Thazha P; Seth, Punit P


    Conjugation of triantennary N-acetyl galactosamine (GalNAc) to oligonucleotide therapeutics results in marked improvement in potency for reducing gene targets expressed in hepatocytes. In this report we describe a robust and efficient solution-phase conjugation strategy to attach triantennary GalNAc clusters (mol. wt. ∼2000) activated as PFP (pentafluorophenyl) esters onto 5'-hexylamino modified antisense oligonucleotides (5'-HA ASOs, mol. wt. ∼8000 Da). The conjugation reaction is efficient and was used to prepare GalNAc conjugated ASOs from milligram to multigram scale. The solution phase method avoids loading of GalNAc clusters onto solid-support for automated synthesis and will facilitate evaluation of GalNAc clusters for structure activity relationship (SAR) studies. Furthermore, we show that transfer of the GalNAc cluster from the 3'-end of an ASO to the 5'-end results in improved potency in cells and animals.

  1. Recognition and sensing of low-epitope targets via ternary complexes with oligonucleotides and synthetic receptors (United States)

    Yang, Kyung-Ae; Barbu, Mihaela; Halim, Marlin; Pallavi, Payal; Kim, Benjamin; Kolpashchikov, Dmitry M.; Pecic, Stevan; Taylor, Steven; Worgall, Tilla S.; Stojanovic, Milan N.


    Oligonucleotide-based receptors or aptamers can interact with small molecules, but the ability to achieve high-affinity and specificity of these interactions depends strongly on functional groups or epitopes displayed by the binding targets. Some classes of targets are particularly challenging: for example, monosaccharides have scarce functionalities and no aptamers have been reported to recognize, let alone distinguish from each other, glucose and other hexoses. Here we report aptamers that differentiate low-epitope targets such as glucose, fructose or galactose by forming ternary complexes with high-epitope organic receptors for monosaccharides. In a follow-up example, we expand this method to isolate high-affinity oligonucleotides against aromatic amino acids complexed in situ with a nonspecific organometallic receptor. The method is general and enables broad clinical use of aptamers for the detection of small molecules in mix-and-measure assays, as demonstrated by monitoring postprandial waves of phenylalanine in human subjects.

  2. Basic electronic circuits

    CERN Document Server

    Buckley, P M


    In the past, the teaching of electricity and electronics has more often than not been carried out from a theoretical and often highly academic standpoint. Fundamentals and basic concepts have often been presented with no indication of their practical appli­ cations, and all too frequently they have been illustrated by artificially contrived laboratory experiments bearing little relationship to the outside world. The course comes in the form of fourteen fairly open-ended constructional experiments or projects. Each experiment has associated with it a construction exercise and an explanation. The basic idea behind this dual presentation is that the student can embark on each circuit following only the briefest possible instructions and that an open-ended approach is thereby not prejudiced by an initial lengthy encounter with the theory behind the project; this being a sure way to dampen enthusiasm at the outset. As the investigation progresses, questions inevitably arise. Descriptions of the phenomena encounte...

  3. Basics of statistical physics

    CERN Document Server

    Müller-Kirsten, Harald J W


    Statistics links microscopic and macroscopic phenomena, and requires for this reason a large number of microscopic elements like atoms. The results are values of maximum probability or of averaging. This introduction to statistical physics concentrates on the basic principles, and attempts to explain these in simple terms supplemented by numerous examples. These basic principles include the difference between classical and quantum statistics, a priori probabilities as related to degeneracies, the vital aspect of indistinguishability as compared with distinguishability in classical physics, the differences between conserved and non-conserved elements, the different ways of counting arrangements in the three statistics (Maxwell-Boltzmann, Fermi-Dirac, Bose-Einstein), the difference between maximization of the number of arrangements of elements, and averaging in the Darwin-Fowler method. Significant applications to solids, radiation and electrons in metals are treated in separate chapters, as well as Bose-Eins...

  4. Basic linear algebra

    CERN Document Server

    Blyth, T S


    Basic Linear Algebra is a text for first year students leading from concrete examples to abstract theorems, via tutorial-type exercises. More exercises (of the kind a student may expect in examination papers) are grouped at the end of each section. The book covers the most important basics of any first course on linear algebra, explaining the algebra of matrices with applications to analytic geometry, systems of linear equations, difference equations and complex numbers. Linear equations are treated via Hermite normal forms which provides a successful and concrete explanation of the notion of linear independence. Another important highlight is the connection between linear mappings and matrices leading to the change of basis theorem which opens the door to the notion of similarity. This new and revised edition features additional exercises and coverage of Cramer's rule (omitted from the first edition). However, it is the new, extra chapter on computer assistance that will be of particular interest to readers:...

  5. Basic Semiconductor Physics

    CERN Document Server

    Hamaguchi, Chihiro


    This book presents a detailed description of the basic semiconductor physics. The reader is assumed to have a basic command of mathematics and some elementary knowledge of solid state physics. The text covers a wide range of important phenomena in semiconductors, from the simple to the advanced. The reader can understand three different methods of energy band calculations, empirical pseudo-potential, k.p perturbation and tight-binding methods. The effective mass approximation and electron motion in a periodic potential, Boltzmann transport equation and deformation potentials used for full band Monte Carlo simulation are discussed. Experiments and theoretical analysis of cyclotron resonance are discussed in detail because the results are essential to the understanding of semiconductor physics. Optical and transport properties, magneto-transport, two dimensional electron gas transport (HEMT and MOSFET), and quantum transport are reviewed, explaining optical transition, electron phonon interactions, electron mob...

  6. Basics of RF electronics

    CERN Document Server

    Gallo, A


    RF electronics deals with the generation, acquisition and manipulation of high-frequency signals. In particle accelerators signals of this kind are abundant, especially in the RF and beam diagnostics systems. In modern machines the complexity of the electronics assemblies dedicated to RF manipulation, beam diagnostics, and feedbacks is continuously increasing, following the demands for improvement of accelerator performance. However, these systems, and in particular their front-ends and back-ends, still rely on well-established basic hardware components and techniques, while down-converted and acquired signals are digitally processed exploiting the rapidly growing computational capability offered by the available technology. This lecture reviews the operational principles of the basic building blocks used for the treatment of high-frequency signals. Devices such as mixers, phase and amplitude detectors, modulators, filters, switches, directional couplers, oscillators, amplifiers, attenuators, and others are d...

  7. Basic plasma physics

    CERN Document Server

    Ghosh, Basudev


    Basic Plasma Physics is designed to serve as an introductory compact textbook for advanced undergraduate, postgraduate and research students taking plasma physics as one of their subject of study for the first time. It covers the current syllabus of plasma physics offered by the most universities and technical institutions. The book requires no background in plasma physics but only elementary knowledge of basic physics and mathematics. Emphasis has been given on the analytical approach. Topics are developed from first principle so that the students can learn through self-study. One chapter has been devoted to describe some practical aspects of plasma physics. Each chapter contains a good number of solved and unsolved problems and a variety of review questions, mostly taken from recent examination papers. Some classroom experiments described in the book will surely help students as well as instructors.

  8. Emulsion Science Basic Principles

    CERN Document Server

    Leal-Calderon, Fernando; Schmitt, Véronique


    Emulsions are generally made out of two immiscible fluids like oil and water, one being dispersed in the second in the presence of surface-active compounds.They are used as intermediate or end products in a huge range of areas including the food, chemical, cosmetic, pharmaceutical, paint, and coating industries. Besides the broad domain of technological interest, emulsions are raising a variety of fundamental questions at the frontier between physics and chemistry. This book aims to give an overview of the most recent advances in emulsion science. The basic principles, covering aspects of emulsions from their preparation to their destruction, are presented in close relation to both the fundamental physics and the applications of these materials. The book is intended to help scientists and engineers in formulating new materials by giving them the basics of emulsion science.

  9. Crystallization of a member of the recFOR DNA repair pathway, RecO, with and without bound oligonucleotide

    Energy Technology Data Exchange (ETDEWEB)

    Aono, Shelly; Hartsch, Thomas; Schulze-Gahmen, Ursula


    RecFOR proteins are important for DNA repair by homologous recombination in bacteria. The RecO protein from Thermus thermophilus was cloned, purified and characterized for its binding to oligonucleotides. The protein was crystallized alone and in complex with a 14-mer oligonucleotide. Both crystal forms grow under different crystallization conditions in the same space group, P3121 or P3221, with almost identical unit cell parameters. Complete data sets were collected to 2.8 Angstrom and 2.5 Angstrom for RecO alone and the RecO-oligonucleotide complex, respectively. Visual comparison of the diffraction patterns between the two crystal forms and calculation of an Rmerge of 33.9 percent on F indicate that one of the crystal forms is indeed a complex of RecO with bound oligonucleotide.

  10. Interfacing click chemistry with automated oligonucleotide synthesis for the preparation of fluorescent DNA probes containing internal xanthene and cyanine dyes

    DEFF Research Database (Denmark)

    Astakhova, I Kira; Wengel, Jesper


    Double-labeled oligonucleotide probes containing fluorophores interacting by energy-transfer mechanisms are essential for modern bioanalysis, molecular diagnostics, and in vivo imaging techniques. Although bright xanthene and cyanine dyes are gaining increased prominence within these fields, litt...

  11. Menstrual Cycle: Basic Biology



    The basic biology of the menstrual cycle is a complex, coordinated sequence of events involving the hypothalamus, anterior pituitary, ovary, and endometrium. The menstrual cycle with all its complexities can be easily perturbed by environmental factors such as stress, extreme exercise, eating disorders, and obesity. Furthermore, genetic influences such as fragile X premutations (Chapter X), X chromosome abnormalities (Chapter X), and galactose-1-phosphate uridyltransferase (GALT) point mutati...

  12. Risk communication basics

    Energy Technology Data Exchange (ETDEWEB)

    Corrado, P.G. [Lawrence Livermore National Laboratory, CA (United States)


    In low-trust, high-concern situations, 50% of your credibility comes from perceived empathy and caring, demonstrated in the first 30 s you come in contact with someone. There is no second chance for a first impression. These and other principles contained in this paper provide you with a basic level of understanding of risk communication. The principles identified are time-tested caveats and will assist you in effectively communicating technical information.

  13. Visual Basic educational programme


    Pranaitis, Arūnas


    Visual basic educational programme Informational Technologies has become such a popular subject that they are applied in all works of life. However, Informational Technologies are still rarely used in the lessons at school. There are such reasons of the mentioned issue: · Insufficient base of computers, · The old software and its disadvantages, · The lack of computerized educational programmes. The aim of the work was to prove that it is actual to create computerized educat...

  14. Basics of Computer Networking

    CERN Document Server

    Robertazzi, Thomas


    Springer Brief Basics of Computer Networking provides a non-mathematical introduction to the world of networks. This book covers both technology for wired and wireless networks. Coverage includes transmission media, local area networks, wide area networks, and network security. Written in a very accessible style for the interested layman by the author of a widely used textbook with many years of experience explaining concepts to the beginner.

  15. Thermodynamics - basic conception

    Energy Technology Data Exchange (ETDEWEB)

    Wee, Eul Bok


    This book tells of basic conception of thermodynamics, condition and property of matter, work and power, thermal efficiency, the principle of the conservation of energy, relationship between work and heat, enthalpy, Jouel's law, complete gasification, the second low of thermodynamics such as thermal efficiency and quality factor, carnot cycle, and entropy, condensation of gas like press of internal combustion engine, vapor, steam power plant and structure, internal combustion cycle, freeze cycle, flow of fluid, combustion and heat transfer.

  16. Decision support basics

    CERN Document Server

    Power, Daniel J


    This book is targeted to busy managers and MBA students who need to grasp the basics of computerized decision support. Some of the topics covered include: What is a DSS? What do managers need to know about computerized decision support? And how can managers identify opportunities to create innovative DSS? Overall the book addresses 35 fundamental questions that are relevant to understanding computerized decision support.

  17. The basic anaesthesia machine. (United States)

    Gurudatt, Cl


    After WTG Morton's first public demonstration in 1846 of use of ether as an anaesthetic agent, for many years anaesthesiologists did not require a machine to deliver anaesthesia to the patients. After the introduction of oxygen and nitrous oxide in the form of compressed gases in cylinders, there was a necessity for mounting these cylinders on a metal frame. This stimulated many people to attempt to construct the anaesthesia machine. HEG Boyle in the year 1917 modified the Gwathmey's machine and this became popular as Boyle anaesthesia machine. Though a lot of changes have been made for the original Boyle machine still the basic structure remains the same. All the subsequent changes which have been brought are mainly to improve the safety of the patients. Knowing the details of the basic machine will make the trainee to understand the additional improvements. It is also important for every practicing anaesthesiologist to have a thorough knowledge of the basic anaesthesia machine for safe conduct of anaesthesia.

  18. Quantum spin model fitting the Yule distribution of oligonucleotides in DNA

    CERN Document Server

    Minichini, C


    A quantum spin chain is identified by the labels of a vector state of a Kashiwara crystal basis. The intensity of the one-spin flip is assumed to depend from the variation of the labels. The rank ordered plot of the numerically computed, averaged in time, transition probabilities is nicely fitted by a Yule distribution, which is the observed distribution of the ranked short oligonucleotides frequency in DNA.

  19. An efficient reagent for the phosphorylation of deoxyribonucleosides, DNA oligonucleotides, and their thermolytic analogues. (United States)

    Ausín, Cristina; Grajkowski, Andrzej; Cieślak, Jacek; Beaucage, Serge L


    [reaction: see text] The phosphoramidite 11 was prepared in three steps from methyl 2-mercaptoacetate and demonstrated efficiency in the synthesis of conventional 5'-/3'-phosphate/thiophosphate monoester derivatives of 2'-deoxyribonucleosides and DNA oligonucleotides. Moreover, the use of 11 has enabled the preparation of the dinucleoside phosphorothioate analogue 26 in high yields (>95%) with minimal cleavage (<2%) of the thermolytic thiophosphate protecting group.

  20. Advances in Antisense Oligonucleotide Development for Target Identification, Validation, and as Novel Therapeutics

    Directory of Open Access Journals (Sweden)

    Moizza Mansoor


    Full Text Available Antisense oligonucleotides (As-ODNs are single stranded, synthetically prepared strands of deoxynucleotide sequences, usually 18–21 nucleotides in length, complementary to the mRNA sequence of the target gene. As-ODNs are able to selectively bind cognate mRNA sequences by sequence-specific hybridization. This results in cleavage or disablement of the mRNA and, thus, inhibits the expression of the target gene. The specificity of the As approach is based on the probability that, in the human genome, any sequence longer than a minimal number of nucleotides (nt, 13 for RNA and 17 for DNA, normally occurs only once. The potential applications of As-ODNs are numerous because mRNA is ubiquitous and is more accessible to manipulation than DNA. With the publication of the human genome sequence, it has become theoretically possible to inhibit mRNA of almost any gene by As-ODNs, in order to get a better understanding of gene function, investigate its role in disease pathology and to study novel therapeutic targets for the diseases caused by dysregulated gene expression. The conceptual simplicity, the availability of gene sequence information from the human genome, the inexpensive availability of synthetic oligonucleotides and the possibility of rational drug design makes As-ODNs powerful tools for target identification, validation and therapeutic intervention. In this review we discuss the latest developments in antisense oligonucleotide design, delivery, pharmacokinetics and potential side effects, as well as its uses in target identification and validation, and finally focus on the current developments of antisense oligonucleotides in therapeutic intervention in various diseases.

  1. Transcript copy number estimation using a mouse whole-genome oligonucleotide microarray


    Carter, Mark G.; Sharov, Alexei A; VanBuren, Vincent; Dudekula, Dawood B.; Carmack, Condie E; Nelson, Charlie; Ko, Minoru SH


    The ability to quantitatively measure the expression of all genes in a given tissue or cell with a single assay is an exciting promise of gene-expression profiling technology. An in situ-synthesized 60-mer oligonucleotide microarray designed to detect transcripts from all mouse genes was validated, as well as a set of exogenous RNA controls derived from the yeast genome (made freely available without restriction), which allow quantitative estimation of absolute endogenous transcript abundance.

  2. Transcript copy number estimation using a mouse whole-genome oligonucleotide microarray (United States)

    Carter, Mark G; Sharov, Alexei A; VanBuren, Vincent; Dudekula, Dawood B; Carmack, Condie E; Nelson, Charlie; Ko, Minoru SH


    The ability to quantitatively measure the expression of all genes in a given tissue or cell with a single assay is an exciting promise of gene-expression profiling technology. An in situ-synthesized 60-mer oligonucleotide microarray designed to detect transcripts from all mouse genes was validated, as well as a set of exogenous RNA controls derived from the yeast genome (made freely available without restriction), which allow quantitative estimation of absolute endogenous transcript abundance. PMID:15998450

  3. Polyimidazole conjugated oligonucleotides reach the nucleus of HeLa cells. (United States)

    Morvan, F; Castex, C; Vivès, E; Imbach, J L


    Oligonucleotide models bearing 6, 12 or 18 histamine residues were synthesized on solid support and labeled with fluorescein. Only the oligo with 6 histamine residues showed a high uptake in HeLa cells with a nuclear localization. Experiment a 4 degrees C or with bafilomicyn A1 suggest that uptake proceeded by an endocytosis mechanism followed by a destabilization of the membrane. Once in the cytoplasm the oligo reached rapidly the nucleus.

  4. Direct Mutagenesis of Thousands of Genomic Targets using Microarray-derived Oligonucleotides

    DEFF Research Database (Denmark)

    Bonde, Mads; Kosuri, Sriram; Genee, Hans Jasper;


    Multiplex Automated Genome Engineering (MAGE) allows simultaneous mutagenesis of multiple target sites in bacterial genomes using short oligonucleotides. However, large-scale mutagenesis requires hundreds to thousands of unique oligos, which are costly to synthesize and impossible to scale-up by ...... insertions per cell. MO-MAGE enables cost-effective large-scale targeted genome engineering that should be useful for a variety of applications in synthetic biology and metabolic engineering....

  5. Modular construction of plasmids through ligation-free assembly of vector components with oligonucleotide linkers. (United States)

    Vroom, Jonathan A; Wang, Clifford L


    We have developed a modular method of plasmid construction that can join multiple DNA components in a single reaction. A nicking enzyme is used to create 5' and 3' overhangs on PCR-generated DNA components. Without the use of ligase or restriction enzymes, components are joined using oligonucleotide linkers that recognize the overhangs. By specifying the sequences of the linkers, desired components can be assembled in any combination and order to generate different plasmid vectors.

  6. Rapid identification of allergenic and pathogenic molds in environmental air by an oligonucleotide array



    Abstract Background Airborne fungi play an important role in causing allergy and infections in susceptible people. Identification of these fungi, based on morphological characteristics, is time-consuming, expertise-demanding, and could be inaccurate. Methods We developed an oligonucleotide array that could accurately identify 21 important airborne fungi (13 genera) that may cause adverse health problems. The method consisted of PCR amplification of the internal transcribed spacer (ITS) region...

  7. Purification of noncoding RNA and bound proteins using FLAG peptide-conjugated antisense-oligonucleotides. (United States)

    Adachi, Shungo; Natsume, Tohru


    To understand the function of certain RNAs, including noncoding RNAs, it is important to identify the proteins that interact with the RNAs. Here we describe the method for purification of ribonucleoprotein (RNP) complexes composed of specific cellular RNAs by pull-down with FLAG peptide-conjugated antisense oligonucleotide (ASO). Using this method, we identified a novel protein component of U7 snRNP complex.

  8. Intrinsically Labeled Fluorescent Oligonucleotide Probes on Quantum Dots for Transduction of Nucleic Acid Hybridization. (United States)

    Shahmuradyan, Anna; Krull, Ulrich J


    Quantum dots (QDs) have been widely used in chemical and biosensing due to their unique photoelectrical properties and are well suited as donors in fluorescence resonance energy transfer (FRET). Selective hybridization interactions of oligonucleotides on QDs have been determined by FRET. Typically, the QD-FRET constructs have made use of labeled targets or have implemented labeled sandwich format assays to introduce dyes in proximity to the QDs for the FRET process. The intention of this new work is to explore a method to incorporate the acceptor dye into the probe molecule. Thiazole orange (TO) derivatives are fluorescent intercalating dyes that have been used for detection of double-stranded nucleic acids. One such dye system has been reported in which single-stranded oligonucleotide probes were doubly labeled with adjacent thiazole orange derivatives. In the absence of the fully complementary (FC) oligonucleotide target, the dyes form an H-aggregate, which results in quenching of fluorescence emission due to excitonic interactions between the dyes. The hybridization of the FC target to the probe provides for dissociation of the aggregate as the dyes intercalate into the double stranded duplex, resulting in increased fluorescence. This work reports investigation of the dependence of the ratiometric signal on the type of linkage used to conjugate the dyes to the probe, the location of the dye along the length of the probe, and the distance between adjacent dye molecules. The limit of detection for 34mer and 90mer targets was found to be identical and was 10 nM (2 pmol), similar to analogous QD-FRET using labeled oligonucleotide target. The detection system could discriminate a one base pair mismatch (1BPM) target and was functional without substantial compromise of the signal in 75% serum. The 1BPM was found to reduce background signal, indicating that the structure of the mismatch affected the environment of the intercalating dyes.

  9. Near-infrared silver cluster optically signaling oligonucleotide hybridization and assembling two DNA hosts. (United States)

    Petty, Jeffrey T; Nicholson, David A; Sergev, Orlin O; Graham, Stuart K


    Silver clusters with ~10 atoms form within DNA strands, and the conjugates are chemical sensors. The DNA host hybridizes with short oligonucleotides, and the cluster moieties optically respond to these analytes. Our studies focus on how the cluster adducts perturb the structure of their DNA hosts. Our sensor is comprised of an oligonucleotide with two components: a 5'-cluster domain that complexes silver clusters and a 3'-recognition site that hybridizes with a target oligonucleotide. The single-stranded sensor encapsulates an ~11 silver atom cluster with violet absorption at 400 nm and with minimal emission. The recognition site hybridizes with complementary oligonucleotides, and the violet cluster converts to an emissive near-infrared cluster with absorption at 730 nm. Our key finding is that the near-infrared cluster coordinates two of its hybridized hosts. The resulting tertiary structure was investigated using intermolecular and intramolecular variants of the same dimer. The intermolecular dimer assembles in concentrated (~5 μM) DNA solutions. Strand stoichiometries and orientations were chromatographically determined using thymine-modified complements that increase the overall conjugate size. The intramolecular dimer develops within a DNA scaffold that is founded on three linked duplexes. The high local cluster concentrations and relative strand arrangements again favor the antiparallel dimer for the near-infrared cluster. When the two monomeric DNA/violet cluster conjugates transform to one dimeric DNA/near-infrared conjugate, the DNA strands accumulate silver. We propose that these correlated changes in DNA structure and silver stoichiometry underlie the violet to near-infrared cluster transformation.

  10. Genome dynamics of short oligonucleotides: the example of bacterial DNA uptake enhancing sequences.

    Directory of Open Access Journals (Sweden)

    Mohammed Bakkali

    Full Text Available Among the many bacteria naturally competent for transformation by DNA uptake-a phenomenon with significant clinical and financial implications- Pasteurellaceae and Neisseriaceae species preferentially take up DNA containing specific short sequences. The genomic overrepresentation of these DNA uptake enhancing sequences (DUES causes preferential uptake of conspecific DNA, but the function(s behind this overrepresentation and its evolution are still a matter for discovery. Here I analyze DUES genome dynamics and evolution and test the validity of the results to other selectively constrained oligonucleotides. I use statistical methods and computer simulations to examine DUESs accumulation in Haemophilus influenzae and Neisseria gonorrhoeae genomes. I analyze DUESs sequence and nucleotide frequencies, as well as those of all their mismatched forms, and prove the dependence of DUESs genomic overrepresentation on their preferential uptake by quantifying and correlating both characteristics. I then argue that mutation, uptake bias, and weak selection against DUESs in less constrained parts of the genome combined are sufficient enough to cause DUESs accumulation in susceptible parts of the genome with no need for other DUES function. The distribution of overrepresentation values across sequences with different mismatch loads compared to the DUES suggests a gradual yet not linear molecular drive of DNA sequences depending on their similarity to the DUES. Other genomically overrepresented sequences, both pro- and eukaryotic, show similar distribution of frequencies suggesting that the molecular drive reported above applies to other frequent oligonucleotides. Rare oligonucleotides, however, seem to be gradually drawn to genomic underrepresentation, thus, suggesting a molecular drag. To my knowledge this work provides the first clear evidence of the gradual evolution of selectively constrained oligonucleotides, including repeated, palindromic and protein

  11. Electrochemical Detection of a Dengue-related Oligonucleotide Sequence Using Ferrocenium as a Hybridization Indicator


    José Luiz de Lima-Filho; Duarte Miguel França dos Prazeres; ernando Rodrigues Ribeiro Teles


    A simple method for electrochemical detection of a synthetic 20-bp oligonucleotide sequence related with dengue virus genome was developed. A complimentary DNA probe sequence was electrostatically immobilized onto a glassy carbon electrode modified with chitosan. Electrochemical detection of hybridization between probe and target was performed by cyclic voltammetry, using ferrocene (Fc+) as a hybridization label. After hybridization, the peak current response of Fc+ oxidation increased around...

  12. Delivering Antisense Morpholino Oligonucleotides to Target Telomerase Splice Variants in Human Embryonic Stem Cells. (United States)

    Radan, Lida; Hughes, Chris S; Teichroeb, Jonathan H; Postovit, Lynne-Marie; Betts, Dean H


    Morpholino oligonucleotides (MO) are an innovative tool that provides a means for examining and modifying gene expression outcomes by antisense interaction with targeted RNA transcripts. The site-specific nature of their binding facilitates focused modulation to alter splice variant expression patterns. Here we describe the steric-blocking of human telomerase reverse transcriptase (hTERT) Δα and Δβ splice variants using MO to examine cellular outcomes related to pluripotency and differentiation in human embryonic stem cells.

  13. A General Tank Test of a Model of the Hull of the British Singapore IIC Flying Boat (United States)

    Dawson, John R; Truscott, Starr


    A general test was made in the N.A.C.A. tank of a 1/12-size model of the hull of the British Singapore IIC flying boat loaned by the Director of Research, British Air Ministry. The results are given in charts and are compared with the results of tests of a model of an American flying-boat hull, the Sikorsky S-40. The Singapore hull has a greater hump resistance but a much lower high-speed resistance than the S-40.


    Institute of Scientific and Technical Information of China (English)

    王美琴; 白春学; 钮善福; 方晓惠; 陈常庆; 陈波


    To explore the effect of antisense oligonucleotide on the production of IL-5 by mouse spleen T lymphocytes.Methods Based on the IL-5 cDNA sequence of mouse, a segment of antisense oligonucleotide was designed and synthesized. 5’-labeling of antisense oligonucleotide was signed by T4 PNK in order that the efficiency of stearylamine liposome in transfecting antisense oligonucleotide can be evaluated. Asthma model was duplicated with ovalbumin(OVA) absorbed to aluminum hydroxide. T lymphocytes of mice were separated by nylon fiber method, then T lymphocytes transfected with different concentration of antisense oligonucleotide with cation stearylamine liposme were incubated respectively in order to observe the effect of antisense oligonucleotide on Il-5 production by T lymphocytes. IL-5 levels in the supernatants of T lymphocyte cultures were determined by ELISA.Results Stearylamine liposome could markedly increase the efficiency of antisense oligonucleotide transfection. The transfection efficiency of antisense oligouncleotide increased approximately 12 times at a ratio of 1: 15m/m (antisense oligonucleotide to SA liposome). In healthy and asthma Balb/c mice, IL-5 was not detectable in the supernatants of T lymphocyte cultures without stimulated with OVA; however, IL-5 was increased markedly in the supernatants of T lymphocyte cultures stimulated with OVA. After transfection with different concentrations of antisense oligonucleotide, IL-5 levels in the supernatants of T lymphocyte cultures were significantly lower than those in control cultured without antisense oligonucleotide transfection. IL-5 levels decreased from 44.60±6.23 pg/ml to 30.70±7.362 pg/ml, 17.20±6.181 pg/ml and 8.16±2.34 pg/ml respectively. And IL-5 synthesis was inhibited by 31.17%, 61.43% and 81.7% respectively.Conclusion IL-5 synthesis could be obviously inhibited by antisense oligonucleotide and showed a markedly correlation between dose and effectiveness. It suggests the production

  15. Development and evaluation of a boat-mounted RFID antenna for monitoring freshwater mussels (United States)

    Fischer, Jesse R.; Neebling, Travis E.; Quist, Michael C.


    Development of radio frequency identification (RFID) technology and passive integrated transponder (PIT) tags has substantially increased the ability of researchers and managers to monitor populations of aquatic organisms. However, use of transportable RFID antenna systems (i.e., backpack-mounted) is currently limited to wadeable aquatic environments (RFID antenna to detect individually PIT-tagged benthic aquatic organisms (mussels). We evaluated the effects of tag orientation on detection distances in water with a 32-mm half-duplex PIT tag. Detection distances up to 50 cm from the antenna coils were obtained, but detection distance was dependent on tag orientation. We also evaluated detection distance of PIT tags beneath the sediment to simulate detection of burrowing mussels with 23- and 32-mm tags. In sand substrate, the maximum detection distance varied from 3.5 cm and 4.5 cm (vertical tag orientation) to 24.7 cm and 39.4 cm (45° tag orientation) for the 23- and 32-mm PIT tags, respectively. Our results suggest a 1.4-m total detection width for tagged mussels on the substrate surface by the boat-mounted antenna system regardless of tag orientation. However, burrowed mussels may require multiple passes to increase detection that would be influenced by depth, tag orientation, and tag size. Construction of the boat-mounted antenna was relatively low in cost (<500 USD) and had several advantages (less labor and time intensive, increased safety) over traditional mussel sampling techniques (diving, snorkeling) in nonwadeable habitats.

  16. An Energy Management System of a Fuel Cell/Battery Hybrid Boat

    Directory of Open Access Journals (Sweden)

    Jingang Han


    Full Text Available All-electric ships are now a standard offering for energy/propulsion systems in boats. In this context, integrating fuel cells (FCs as power sources in hybrid energy systems can be an interesting solution because of their high efficiency and low emission. The energy management strategy for different power sources has a great influence on the fuel consumption, dynamic performance and service life of these power sources. This paper presents a hybrid FC/battery power system for a low power boat. The hybrid system consists of the association of a proton exchange membrane fuel cell (PEMFC and battery bank. The mathematical models for the components of the hybrid system are presented. These models are implemented in Matlab/Simulink environment. Simulations allow analyzing the dynamic performance and power allocation according to a typical driving cycle. In this system, an efficient energy management system (EMS based on operation states is proposed. This EMS strategy determines the operating point of each component of the system in order to maximize the system efficiency. Simulation results validate the adequacy of the hybrid power system and the proposed EMS for real ship driving cycles.

  17. Hydrodynamics of a self-propelled camphor boat at the air-water interface (United States)

    Akella, Sathish; Singh, Dhiraj; Singh, Ravi; Bandi, Mahesh


    A camphor tablet, when placed at the air-water interface undergoes sublimation and camphor vapour spreads radially outwards across the surface due to Marangoni forces. This steady camphor influx from tablet onto the air-water interface is balanced by the camphor outflux due to evaporation. When spontaneous fluctuations in evaporation break the axial symmetry of Marangoni force acting radially outwards, the camphor tablet is propelled like a boat along the water surface. We report experiments on the hydrodynamics of a self-propelled camphor boat at air-water interfaces. We observe three different modes of motion, namely continuous, harmonic and periodic, due to the volatile nature of camphor. We explain these modes in terms of ratio of two time-scales: the time-scale over which viscous forces are dominant over the Marangoni forces (τη) and the time-scale over which Marangoni forces are dominant over the viscous forces (τσ). The continuous, harmonic and periodic motions are observed when τη /τσ ~ 1 , τη /τσ >= 1 and τη /τσ >> 1 respectively. Experimentally, the ratio of the time scales is varied by changing the interfacial tension of the air-water interface using Sodium Dodecyl Sulfate. This work was supported by the Collective Interactions Unit, OIST Graduate University.

  18. Energetic analyses of chair and boat conformations of maleimide substituted cyclohexane derivatives. (United States)

    Remmert, Sarah; Parish, Carol


    An analysis of the conformational preferences of two maleimide substituted cyclohexane derivatives proposed as scaffolds for HIV-1 fusion inhibitors is presented. Hybrid Low Mode-Monte Carlo (1:1) conformational searches using seven different force fields were performed in combination with the GBSA(water) solvent model. Low energy structures identified in this way were subjected to geometry optimization on the B3LYP/6-31G** surface. Solvent effects were included in the quantum calculation using the self-consistent reaction field model for water. Quantum results indicate that the 1,3,5-maleimide functionalized 1,3,5-methyl cyclohexane is more stable in the boat conformation, whereas 1,3,5-maleimide functionalized cyclohexane adopts the expected chair conformation with equatorial arms. None of the force fields studied was able to predict the unexpected preference for the boat conformation of 1,3,5-maleimide functionalized 1,3,5-methyl cyclohexane. Comparison of low energy and experimental structures was also performed.

  19. Conformation of hindered piperidines: Spectroscopic evidence for contribution of boat conformations

    Indian Academy of Sciences (India)

    A Thangamani; J Jayabharathi; A Manimekalai


    High resolution 1H and 13C NMR resonance assignments and conformational assignments were carried out for four (3)-benzyl-(2),(6)-bis(aryl)piperidin-4-ones 1-4 and their four -nitroso-(3)-benzyl-(2),(6)-bis(aryl)piperidin-4-ones 5-8. In addition to conventional 1D NMR methods, 2D shiftcorrelated NMR techniques (1H-1H COSY and 1H-13C COSY) were used for signal assignments. At room temperature the (3)-benzyl-(2),(6)-bis(aryl)piperidin-4-ones 1-4 exist in only one isomeric form whereas their -nitroso derivatives 5-8 exist in two isomeric forms. The preferred conformations of both the isomeric forms of nitrosamines were determined by comparison of the spectral data with those of the corresponding parent amines 1-4 and with the aid of substituent parameters. The results indicate the presence of an equilibrium mixture of boat forms B1 and B2 for Z isomers of 5-8. For the E isomers of 5-8, boat form B1 is predicted to be the major conformer. The piperidin-4-ones 1-4 exist in normal chair conformations with equatorial orientations of all the substituents.

  20. Synthesis of a multibranched porphyrin-oligonucleotide scaffold for the construction of DNA-based nano-architectures. (United States)

    Clavé, Guillaume; Chatelain, Grégory; Filoramo, Arianna; Gasparutto, Didier; Saint-Pierre, Christine; Le Cam, Eric; Piétrement, Olivier; Guérineau, Vincent; Campidelli, Stéphane


    The interest in the functionalization of oligonucleotides with organic molecules has grown considerably over the last decade. In this work, we report on the synthesis and characterization of porphyrin-oligonucleotide hybrids containing one to four DNA strands (P1-P4). The hybrid P4, which inserts one porphyrin and four DNA fragments, was combined with gold nanoparticles and imaged by transmission electron microscopy.

  1. Thermolytic CpG-containing DNA oligonucleotides as potential immunotherapeutic prodrugs. (United States)

    Grajkowski, Andrzej; Pedras-Vasconcelos, Joao; Wang, Vivian; Ausín, Cristina; Hess, Sonja; Verthelyi, Daniela; Beaucage, Serge L


    A CpG-containing DNA oligonucleotide functionalized with the 2-(N-formyl-N-methyl)aminoethyl thiophosphate protecting group (CpG ODN fma1555) was prepared from phosphoramidites 1a-d using solid-phase techniques. The oligonucleotide behaved as a prodrug by virtue of its conversion to the well-studied immunomodulatory CpG ODN 1555 through thermolytic cleavage of the 2-(N-formyl-N-methyl)aminoethyl thiophosphate protecting group. Such a conversion occurred at 37 degrees C with a half-time of 73 h. The immunostimulatory properties of CpG ODN fma1555 were evaluated in two in vivo assays, one of which consisted of mice challenged in the ear with live Leishmania major metacyclic promastigotes. Local intradermal administration of CpG ODN fma1555 was as effective as that of CpG ODN 1555 in reducing the size of Leishmania lesions over time. In a different infectious model, CpG ODN 1555 prevented the death of Tacaribe-infected mice (43% survival) when administered between day 0 and 3 post infection. Administration of CpG ODN fma1555 three days before infection resulted in improved immunoprotection (60-70% survival). Moreover, co-administration of CpG ODN fma1555 and CpG ODN 1555 in this model increased the window for therapeutic treatment against Tacaribe virus infection, and thus supports the use of thermolytic oligonucleotides as prodrugs in the effective treatment of infectious diseases.

  2. Sequence-specific targeting of RNA with an oligonucleotide-neomycin conjugate. (United States)

    Charles, Irudayasamy; Xi, Hongjuan; Arya, Dev P


    The synthesis of neomycin covalently attached at the C5-position of 2'-deoxyuridine is reported. The synthesis outlined allows for incorporation of an aminoglycoside (neomycin) at any given site in an oligonucleotide (ODN) where a thymidine (or uridine) is present. Incorporation of this modified base into an oligonucleotide, which is complementary to a seven-bases-long alpha-sarcin loop RNA sequence, leads to enhanced duplex hybridization. The increase in Tm for this duplex (DeltaTm = 6 degrees C) suggests a favorable interaction of neomycin within the duplex groove. CD spectroscopy shows that the modified duplex adopts an A-type confirmation. ITC measurements indicate the additive effects of ODN and neomycin binding to the RNA target (Ka = 4.5 x 107 M-1). The enhanced stability of the hybrid duplex from this neomycin-ODN conjugate originates primarily from the enthalpic contribution of neomycin {DeltaDeltaHobs = -7.21 kcal/mol (DeltaHneomycin conjugated - DeltaH nonconjugated)} binding to the hybrid duplex. The short linker length allows for selective stabilization of the hybrid duplex over the hybrid triplex. The results described here open up new avenues in the design and synthesis of nucleo-aminoglycoside-conjugates (N-Ag-C) where the inclusion of any number of aminoglycoside (neomycin) molecules per oligonucleotide can be accomplished.

  3. Pericellular matrix formation alters the efficiency of intracellular uptake of oligonucleotides in osteosarcoma cells. (United States)

    Suzuki, Yoshitaka; Nishida, Yoshihiro; Naruse, Takahiro; Gemba, Takefumi; Ishiguro, Naoki


    One of the crucial roles of tumor extracellular matrix is to act as a barrier to drug delivery. In this study, we analyzed the relationship between the formation of tumor extracellular matrix and the efficiency of intracellular uptake of oligonucleotides in human osteosarcoma cell lines, HOS, and MG-63. Oligonucleotides used in this study were nuclear factor-kappa B (NF-kappaB) decoy, which might be a therapeutic tool for neoplasms. Pericellular matrix formation was examined by particle exclusion assay. Cellular uptake of fluorescein isothiocyanate-labeled NF-kappaB decoy was evaluated by fluorescent microscopy and flow cytometry. Effects of NF-kappaB decoy on cell viability and cell cycle arrest in MG-63 cells were determined by MTT assay and flow cytometry, respectively. MG-63 cells exhibited abundant pericellular matrix with time compared with HOS cells. Uptake of fluorescein isothiocyanate-labeled NF-kappaB decoy decreased in MG-63 cells with time but not in HOS cells in both monolayer and three-dimensional culture using matrigel. However, after enzymatic removal of pericellular matrix, the uptake markedly recovered in MG-63 cells. NF-kappaB decoy inhibited cell proliferation and induced G0/G1 cell cycle arrest in MG-63 cells. These results suggest that abundant pericellular matrix might disturb the uptake of NF-kappaB decoy, and modification of pericellular matrix composition would increase the efficacy of exogenous oligonucleotides treatment for neoplasms.

  4. Detection of Glucose with Atomic Absorption Spectroscopy by Using Oligonucleotide Functionalized Gold Nanoparticle. (United States)

    Zhang, Hong; Yan, Honglian; Ling, Liansheng


    A novel method for the detection of glucose was established with atomic absorption spectroscopy by using the label of gold nanoparticle (AuNP). Silver-coated glass assembled with oligonucleotide 5'-SH-T12-AGA CAA GAG AGG-3' (Oligo 1) was acted as separation probe, oligonucleotide 5'-CAA CAG AGA ACG-T12-SH-3' modified gold nanoparticle (AuNP-Oligo 2) was acted as signal-reporting probe. Oligonucleotide 5'-CGT TCT CTG TTG CCT CTC TTG TCT-3' (Oligo 3) could hybridize with Oligo 1 on the surface of silver-coated glass and AuNP-Oligo 2, and free AuNP-Oligo 2 could be removed by rinsing with buffer. Hence the concentration of Oligo 3 was transformed into the concentration of gold element. In addition, Oligo 3 could be cleaved into DNA fragments by glucose, glucose oxidase and Fe(2+)-EDTA through Fenton reaction. Thereby the concentration of glucose could be transformed to the absorbance of gold element. Under the optimum conditions, the integrated absorbance decreased proportionally to the concentration of glucose over the range from 50.0 μM to 1.0 mM with a detection limit of 40.0 μM. Moreover, satisfactory result was obtained when the assay was used to determinate glucose in human serum.

  5. Membrane-based oligonucleotide array developed from multiple markers for the detection of many Phytophthora species. (United States)

    Chen, Wen; Djama, Zeinab Robleh; Coffey, Michael D; Martin, Frank N; Bilodeau, Guillaume J; Radmer, Lorien; Denton, Geoff; Lévesque, C André


    Most Phytophthora spp. are destructive plant pathogens; therefore, effective monitoring and accurate early detection are important means of preventing potential epidemics and outbreaks of diseases. In the current study, a membrane-based oligonucleotide array was developed that can detect Phytophthora spp. reliably using three DNA regions; namely, the internal transcribed spacer (ITS), the 5' end of cytochrome c oxidase 1 gene (cox1), and the intergenic region between cytochrome c oxidase 2 gene (cox2) and cox1 (cox2-1 spacer). Each sequence data set contained ≈250 sequences representing 98 described and 15 undescribed species of Phytophthora. The array was validated with 143 pure cultures and 35 field samples. Together, nonrejected oligonucleotides from all three markers have the ability to reliably detect 82 described and 8 undescribed Phytophthora spp., including several quarantine or regulated pathogens such as Phytophthora ramorum. Our results showed that a DNA array containing signature oligonucleotides designed from multiple genomic regions provided robustness and redundancy for the detection and differentiation of closely related taxon groups. This array has the potential to be used as a routine diagnostic tool for Phytophthora spp. from complex environmental samples without the need for extensive growth of cultures.

  6. Molecular phylogenetics before sequences: oligonucleotide catalogs as k-mer spectra. (United States)

    Ragan, Mark A; Bernard, Guillaume; Chan, Cheong Xin


    From 1971 to 1985, Carl Woese and colleagues generated oligonucleotide catalogs of 16S/18S rRNAs from more than 400 organisms. Using these incomplete and imperfect data, Carl and his colleagues developed unprecedented insights into the structure, function, and evolution of the large RNA components of the translational apparatus. They recognized a third domain of life, revealed the phylogenetic backbone of bacteria (and its limitations), delineated taxa, and explored the tempo and mode of microbial evolution. For these discoveries to have stood the test of time, oligonucleotide catalogs must carry significant phylogenetic signal; they thus bear re-examination in view of the current interest in alignment-free phylogenetics based on k-mers. Here we consider the aims, successes, and limitations of this early phase of molecular phylogenetics. We computationally generate oligonucleotide sets (e-catalogs) from 16S/18S rRNA sequences, calculate pairwise distances between them based on D 2 statistics, compute distance trees, and compare their performance against alignment-based and k-mer trees. Although the catalogs themselves were superseded by full-length sequences, this stage in the development of computational molecular biology remains instructive for us today.

  7. Efficient delivery of RNA interference oligonucleotides to polarized airway epithelia in vitro. (United States)

    Ramachandran, Shyam; Krishnamurthy, Sateesh; Jacobi, Ashley M; Wohlford-Lenane, Christine; Behlke, Mark A; Davidson, Beverly L; McCray, Paul B


    Polarized and pseudostratified primary airway epithelia present barriers that significantly reduce their transfection efficiency and the efficacy of RNA interference oligonucleotides. This creates an impediment in studies of the airway epithelium, diminishing the utility of loss-of-function as a research tool. Here we outline methods to introduce RNAi oligonucleotides into primary human and porcine airway epithelia grown at an air-liquid interface and difficult-to-transfect transformed epithelial cell lines grown on plastic. At the time of plating, we reverse transfect small-interfering RNA (siRNA), Dicer-substrate siRNA, or microRNA oligonucleotides into cells by use of lipid or peptide transfection reagents. Using this approach we achieve significant knockdown in vitro of hypoxanthine-guanine phosphoribosyltransferase, IL-8, and CFTR expression at the mRNA and protein levels in 1-3 days. We also attain significant reduction of secreted IL-8 in polarized primary pig airway epithelia 3 days posttransfection and inhibition of CFTR-mediated Cl⁻ conductance in polarized air-liquid interface cultures of human airway epithelia 2 wk posttransfection. These results highlight an efficient means to deliver RNA interference reagents to airway epithelial cells and achieve significant knockdown of target gene expression and function. The ability to reliably conduct loss-of-function assays in polarized primary airway epithelia offers benefits to research in studies of epithelial cell homeostasis, candidate gene function, gene-based therapeutics, microRNA biology, and targeting the replication of respiratory viruses.

  8. Antisense oligonucleotides as innovative therapeutic strategy in the treatment of high-grade gliomas. (United States)

    Caruso, Gerardo; Caffo, Mariella; Raudino, Giuseppe; Alafaci, Concetta; Salpietro, Francesco M; Tomasello, Francesco


    Despite the intensive recent research in cancer therapy, the prognosis in patients affected by high-grade gliomas is still very unfavorable. The efficacy of classical anti-cancer strategies is seriously limited by lack of specific therapies against malignant cells. The extracellular matrix plays a pivotal role in processes such as differentiation, apoptosis, and migration in both the normal and the pathologic nervous system. Glial tumors seem to be able to create a favorable environment for the invasion of glioma cells in cerebral parenchyma when they combine with the extracellular matrix via cell surface receptors. Glioma cells synthesize matrix proteins, such as tenascin, laminin, fibronectin that facilitate the tumor cell's motility. New treatments have shown to hit the acting molecules in the tumor growth and to increase the efficacy and minimize the toxicity. Antisense oligonucleotides are synthetic stretches of DNA which hybridize with specific mRNA strands. The specificity of hybridization makes antisense method an interesting strategy to selectively modulate the expression of genes involved in tumorigenesis. In this review we will focus on the mechanisms of action of antisense oligonucleotides and report clinical and experimental studies on the treatment of high-grade gliomas. We will also report the patents of preclinical and/or clinical studies that adopt the antisense oligonucleotide therapy list in cerebral gliomas.

  9. Immunostimulatory oligonucleotide, CpG-like motif exists in Lactobacillus delbrueckii ssp. bulgaricus NIAI B6. (United States)

    Kitazawa, Haruki; Watanabe, Hiroshi; Shimosato, Takeshi; Kawai, Yasushi; Itoh, Takatoshi; Saito, Tadao


    The present study was conducted to find an immunostimulatory oligonucleotide derived from yogurt starter cultures. The chromosomal DNA was purified from nine strains of Lactobacillus delbrueckii ssp. bulgaricus and six strains of Streptococcus thermophilus. An immunostimulatory ability of the DNA was examined in a proliferation of peyer's patch and splenic B cells. Only the DNA from L. bulgaricus NIAI B6 induced a significant proliferation of both cells. When the DNA was cloned and amplified using PCR, the mitogenic activities to B cells were significantly increased by 13 of 135 DNA clones. Ten homologous nucleotide sequences were found as possible oligonucleotide sequences of mitogens, and were then chemically synthesized (sOL-LB1 to sOL-LB10). One CpG-like motif (sOL-LB7; 5'-CGGCACGCTCACGATTCTTG-3') was identified as an immunostimulatory oligonucleotide, but it did not contain palindromic CpG structure known as a B cell-specific mitogen. The sOL-LB7 substantially bound to B cells and increased the CD69 positive cells in peyer's patch cells. This study demonstrated that L. bulgaricus NIAI B6 was a good candidate of a starter culture for the production of new functional foods, "Bio-Defense Foods".

  10. ProbeMaker: an extensible framework for design of sets of oligonucleotide probes

    Directory of Open Access Journals (Sweden)

    Nilsson Mats


    Full Text Available Abstract Background Procedures for genetic analyses based on oligonucleotide probes are powerful tools that can allow highly parallel investigations of genetic material. Such procedures require the design of large sets of probes using application-specific design constraints. Results ProbeMaker is a software framework for computer-assisted design and analysis of sets of oligonucleotide probe sequences. The tool assists in the design of probes for sets of target sequences, incorporating sequence motifs for purposes such as amplification, visualization, or identification. An extension system allows the framework to be equipped with application-specific components for evaluation of probe sequences, and provides the possibility to include support for importing sequence data from a variety of file formats. Conclusion ProbeMaker is a suitable tool for many different oligonucleotide design and analysis tasks, including the design of probe sets for various types of parallel genetic analyses, experimental validation of design parameters, and in silico testing of probe sequence evaluation algorithms.

  11. Efficient inhibition of human telomerase activity by antisense oligonucleotides sensitizes cancer cells to radiotherapy

    Institute of Scientific and Technical Information of China (English)

    Xue-mei JI; Cong-hua XIE; Ming-hao FANG; Fu-xiang ZHOU; Wen-jie ZHANG; Ming-sheng ZHANG; Yun-feng ZHOU


    Aim: To investigate the effect of the antisense oligonucleotides (ASODN) specific for human telomerase RNA (hTR) on radio sensitization and proliferation inhibition in human neurogliocytoma cells (U251). Methods: U251 cells were transfected with hTR ASODN or nonspecific oligonucleotides (NSODN). Before and after irradiation of 60Co-γray, telomerase activity was assayed by telomeric repeat amplification protocol (TRAP-PCR-ELISA), and DNA damage and repair were examined by the comet assay. The classical colony assay was used to plot the cell-survival curve, to detect the D0 value. Results: hTR antisense oligonucleotides could downregulate the telomerase activity, increase radiation induced DNA damage and reduce the subsequent repair. Furthermore, it could inhibit the proliferation and decrease the D0 value which demonstrates rising radiosensitivity. However, telomere length was unchanged over a short period of time. Conclusion: These findings suggest that an ASODN-based strategy may be used to develop telomerase inhibitors, which can efficiently sensitize radiotherapy.

  12. Tanks Test of a Model of the Hull of the Navy PB-1 Flying Boat - N.A.C.A. Model 52 (United States)

    Allison, John M


    A model of the hull of the Navy PB-1 flying boat was tested in the N.A.C.A. tank as part of a program intended to provide information regarding the water performance of hulls of flying boats of earlier design for which hydrodynamic data have heretofore been unavailable. Tests were made according to the general method over the range of practical loadings with the model both fixed in trim and free to trim. A free-to-trim test according to the specific method was also made for the design load and take-off speed corresponding to those of the full-scale flying boat. The resistance obtained from the fixed-trim test was found to be about the same as that of the model of the NC flying-boat hull, and greater at the hump but smaller at high speeds than that of a model of the Sikorsky S-40 flying-boat hull.

  13. Designing a Two-Phase BLDC Motor and Finite-Element Analysis of Designing a Controller in Order to Manoeuvre High-Speed Boats in Marine Turbulences

    Directory of Open Access Journals (Sweden)

    Mehrdad Jafarboland


    Full Text Available High speed boats are small kind of boats which are piloted with boatman.These boats are usually used in strategic commissions like military which fast speeds and good manoeuvrability are of essential importance.Rough sea path, marine turbulences and multi input multi output nonlinear dynamical model of this boats,make it very difficult to pilot and manoeuvre this paper,.two controller are designed and proposed for pursuing desired path and manoeuvring fast speeds.Fuzzy controller is compared with the result of PID controller. Simulation results also indicate that these proposed controllers have suitable responses and can pilot the boat in the best manner.

  14. Discovery and development of the G-rich oligonucleotide AS1411 as a novel treatment for cancer. (United States)

    Bates, Paula J; Laber, Damian A; Miller, Donald M; Thomas, Shelia D; Trent, John O


    Certain guanine-rich (G-rich) DNA and RNA molecules can associate intermolecularly or intramolecularly to form four stranded or "quadruplex" structures, which have unusual biophysical and biological properties. Several synthetic G-rich quadruplex-forming oligodeoxynucleotides have recently been investigated as therapeutic agents for various human diseases. We refer to these biologically active G-rich oligonucleotides as aptamers because their activities arise from binding to protein targets via shape-specific recognition (analogous to antibody-antigen binding). As therapeutic agents, the G-rich aptamers may have some advantages over monoclonal antibodies and other oligonucleotide-based approaches. For example, quadruplex oligonucleotides are non-immunogenic, heat stable and they have increased resistance to serum nucleases and enhanced cellular uptake compared to unstructured sequences. In this review, we describe the characteristics and activities of G-rich oligonucleotides. We also give a personal perspective on the discovery and development of AS1411, an antiproliferative G-rich phosphodiester oligonucleotide that is currently being tested as an anticancer agent in Phase II clinical trials. This molecule functions as an aptamer to nucleolin, a multifunctional protein that is highly expressed by cancer cells, both intracellularly and on the cell surface. Thus, the serendipitous discovery of the G-rich oligonucleotides also led to the identification of nucleolin as a new molecular target for cancer therapy.

  15. Thermolytic 4-methylthio-1-butyl group for phosphate/thiophosphate protection in solid-phase synthesis of DNA oligonucleotides. (United States)

    Cieślak, Jacek; Grajkowski, Andrzej; Livengood, Victor; Beaucage, Serge L


    The thermolabile 4-methylthio-1-butyl phosphate/thiophosphate protecting group for DNA oligonucleotides has been investigated for its potential application to a "heat-driven" process for either oligonucleotide synthesis on diagnostic microarrays or, oppositely, to the large-scale preparation of therapeutic oligonucleotides. The preparation of phosphoramidites 10a-d is straightforward, and the incorporation of these amidites into oligonucleotides via solid-phase techniques proceeds as efficiently as that achieved with 2-cyanoethyl deoxyribonucleoside phosphoramidites. The versatility of the 4-methylthio-1-butyl phosphate/thiophosphate protecting group is exemplified by its facile removal from oligonucleotides upon heating for 30 min at 55 degrees C in an aqueous buffer under neutral conditions or within 2 h at 55 degrees C in concentrated NH(4)OH. The deprotection reaction occurs through an intramolecular cyclodeesterification mechanism leading to the formation of sulfonium salt 18. When mixed with deoxyribonucleosides and N-protected 2'-deoxyribonucleosides or with a model phosphorothioate diester under conditions approximating those of large-scale (>50 mmol) oligonucleotide deprotection reactions, the salt 18 did not significantly alter DNA nucleobases or desulfurize the phosphorothioate diester model to an appreciable extent.

  16. Oligonucleotide and Parylene Surface Coating of Polystyrene and ePTFE for Improved Endothelial Cell Attachment and Hemocompatibility

    Directory of Open Access Journals (Sweden)

    Martina Schleicher


    Full Text Available In vivo self-endothelialization by endothelial cell adhesion on cardiovascular implants is highly desirable. DNA-oligonucleotides are an intriguing coating material with nonimmunogenic characteristics and the feasibility of easy and rapid chemical fabrication. The objective of this study was the creation of cell adhesive DNA-oligonucleotide coatings on vascular implant surfaces. DNA-oligonucleotides immobilized by adsorption on parylene (poly(monoaminomethyl-para-xylene coated polystyrene and ePTFE were resistant to high shear stress (9.5 N/m2 and human blood serum for up to 96 h. Adhesion of murine endothelial progenitor cells, HUVECs and endothelial cells from human adult saphenous veins as well as viability over a period of 14 days of HUVECs on oligonucleotide coated samples under dynamic culture conditions was significantly enhanced (P<0.05. Oligonucleotide-coated surfaces revealed low thrombogenicity and excellent hemocompatibility after incubation with human blood. These properties suggest the suitability of immobilization of DNA-oligonucleotides for biofunctionalization of blood vessel substitutes for improved in vivo endothelialization.

  17. Cross-protective effect of antisense oligonucleotide developed against the common 3' NCR of influenza A virus genome. (United States)

    Kumar, Prashant; Kumar, Binod; Rajput, Roopali; Saxena, Latika; Banerjea, Akhil C; Khanna, Madhu


    The influenza A virus (IAV) has eight segmented single-stranded RNA genome containing a common and evolutionarily conserved non-coding region (NCRs) at 5' and 3' ends that are important for the virus replication. In this study, we designed an antisense oligonucleotide against the 3' NCR of vital segments of the IAV genome to inhibit its replication. The results demonstrated that the co-transfection of Madine Darby Canine Kidney (MDCK) cells with the antisense oligonucleotide and the plasmids encoding the viral genes led to the down-regulation of the viral gene expression. The designed antisense molecules reduced the cytopathic effect caused by A/PR/8/34 (H1N1), A/Udorn/307/72 (H3N2), and A/New Caledonia/20/99 (H1N1) strains of IAV for almost 48 h. Furthermore, the intra-venous delivery of this oligonucleotide significantly reduced the viral titers in the lungs of infected mice and protected the mice from lethal effects of all the strains of influenza virus. The study demonstrated that the antisense oligonucleotide designed against the NCR region inhibits the expression of the viral genome. The decrease of the cytopathic effect in the MDCK cells and increase in survival of mice confirmed the reduction of virus multiplication and pathogenesis in the presence of antisense oligonucleotide. Thus, we demonstrate that a single antisense oligonucleotide is capable of providing protection against more than one strains of the IAV.

  18. Thermolytic release of covalently linked DNA oligonucleotides and their conjugates from controlled-pore glass at near neutral pH. (United States)

    Grajkowski, Andrzej; Cieślak, Jacek; Kauffman, Jon S; Duff, Robert J; Norris, Scott; Freedberg, Darón I; Beaucage, Serge L


    The functionalization of long chain alkylamine controlled-pore glass (CPG) with a 3-hydroxypropyl-(2-cyanoethyl)thiophosphoryl linker and its conversion to the support 7 has led to the synthesis of DNA oligonucleotides and their 3'- or (3',5')-conjugates. Indeed, CPG support 7 has been successfully employed in the synthesis of both native and fully phosphorothioated DNA 20-mers. Unlike conventional succinylated CPG supports, this distinctively functionalized support allows oligonucleotide deprotection and removal of the deprotection side products to proceed without releasing the oligonucleotide into the aqueous milieu. When freed from deprotection side products, the DNA oligonucleotide is thermolytically released from the support within 2 h under nearly neutral conditions (pH 7.2, 90 degrees C). The quality of these oligonucleotides is comparable to that of identical oligonucleotides synthesized from succinylated CPG supports in terms of shorter than full length oligonucleotide contaminants and overall yields. The versatility of the thermolytic CPG support 7 is further demonstrated by the synthesis of a DNA oligonucleotide (20-mer) and its conjugation with an azido and alkynyl groups at both 5'-and 3'-termini, respectively. The functionality of the (3',5')-heteroconjugated oligonucleotide 18 is verified by its circularization to the DNA oligonucleotide 19 under "click" chemistry conditions.

  19. Catalyst in Basic Oleochemicals

    Directory of Open Access Journals (Sweden)

    Eva Suyenty


    Full Text Available Currently Indonesia is the world largest palm oil producer with production volume reaching 16 million tones per annum. The high crude oil and ethylene prices in the last 3 – 4 years contribute to the healthy demand growth for basic oleochemicals: fatty acids and fatty alcohols. Oleochemicals are starting to replace crude oil derived products in various applications. As widely practiced in petrochemical industry, catalyst plays a very important role in the production of basic oleochemicals. Catalytic reactions are abound in the production of oleochemicals: Nickel based catalysts are used in the hydrogenation of unsaturated fatty acids; sodium methylate catalyst in the transesterification of triglycerides; sulfonic based polystyrene resin catalyst in esterification of fatty acids; and copper chromite/copper zinc catalyst in the high pressure hydrogenation of methyl esters or fatty acids to produce fatty alcohols. To maintain long catalyst life, it is crucial to ensure the absence of catalyst poisons and inhibitors in the feed. The preparation methods of nickel and copper chromite catalysts are as follows: precipitation, filtration, drying, and calcinations. Sodium methylate is derived from direct reaction of sodium metal and methanol under inert gas. The sulfonic based polystyrene resin is derived from sulfonation of polystyrene crosslinked with di-vinyl-benzene. © 2007 BCREC UNDIP. All rights reserved.[Presented at Symposium and Congress of MKICS 2007, 18-19 April 2007, Semarang, Indonesia][How to Cite: E. Suyenty, H. Sentosa, M. Agustine, S. Anwar, A. Lie, E. Sutanto. (2007. Catalyst in Basic Oleochemicals. Bulletin of Chemical Reaction Engineering and Catalysis, 2 (2-3: 22-31.  doi:10.9767/bcrec.2.2-3.6.22-31][How to Link/DOI: || or local:

  20. Basic heat transfer

    CERN Document Server

    Bacon, D H


    Basic Heat Transfer aims to help readers use a computer to solve heat transfer problems and to promote greater understanding by changing data values and observing the effects, which are necessary in design and optimization calculations.The book is concerned with applications including insulation and heating in buildings and pipes, temperature distributions in solids for steady state and transient conditions, the determination of surface heat transfer coefficients for convection in various situations, radiation heat transfer in grey body problems, the use of finned surfaces, and simple heat exc