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The Cry (crystal) proteins from Bacillus thuringiensis are known to have toxicity against a variety of insects and have been exploited to control insect pests through transgenic plants and biopesticides. B. thuringiensis SBS BT-1 carrying the cry2 genes was isolated from soil samples in Pakistan. The 2-kb full length cry2Ac gene was cloned, sequenced, and submitted to the EMBL DNA database (Accession No. AM292031). For expression analysis, Escherichia coli DH5α was transformed with the fragment sub-cloned in pET22b expression vector using NdeI and HindIII restriction sites, and later confirmed by restriction endonuclease analysis. To assess the toxicity of Cry2Ac7 protein against lepidopteran and dipteran insects, BL21 (codon plus) strain of E. coli was further transformed with the recombinant plasmid. The 65-kDa protein was expressed in the form of inclusion bodies up to 180 OD units per liter of the medium. Inclusions were washed with a buffer containing 1.5% Triton-X 100 and >90% pure Cry2Ac7 was obtained. The inclusion bodies were dissolved in 50 mM K2CO3 (pH 11.5), dialyzed, and freeze-dried. This freeze-dried protein as well as inclusion bodies were used in bioassays against larvae of Helicoverpa armigera and Musca domestica. The freeze-dried protein was toxic to H. armigera larvae with an LC50 value of 131 ng/mL. However, Cry2Ac7 produced in E. coli did not show any mortality to M. domestica larvae. This is the first report of Cry2Ac protein toxic to H. armigera. PMID:29099767
Gene Ontology (GO) semantic similarity measures are being used for biological knowledge discovery based on GO annotations by integrating biological information contained in the GO structure into data analyses. To empower users to quickly compute, manipulate and explore these measures, we introduce A-DaGO-Fun (ADaptable Gene Ontology semantic similarity-based Functional analysis). It is a portable software package integrating all known GO information content-based semantic similarity measures and relevant biological applications associated with these measures. A-DaGO-Fun has the advantage not only of handling datasets from the current high-throughput genome-wide applications, but also allowing users to choose the most relevant semantic similarity approach for their biological applications and to adapt a given module to their needs. A-DaGO-Fun is freely available to the research community at It is implemented in Linux using Python under free software (GNU General Public Licence). firstname.lastname@example.org or Nicola.Mulder@uct.ac.za Supplementary data are available at Bioinformatics online. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: email@example.com.
Secoisolariciresinol diglucoside (SDG), the main lignan in whole grain flaxseed, is a potent antioxidant and free radical scavenger with known radioprotective properties. However, the exact mechanism of SDG radioprotection is not well understood. The current study identified a novel mechanism of DNA radioprotection by SDG in physiological solutions by scavenging active chlorine species (ACS) and reducing chlorinated nucleobases. The ACS scavenging activity of SDG was determined using two highly specific fluoroprobes: hypochlorite-specific 3'-(p-aminophenyl) fluorescein (APF) and hydroxyl radical-sensitive 3'-(p-hydroxyphenyl) fluorescein (HPF). Dopamine, an SDG structural analog, was used for proton (1)H NMR studies to trap primary ACS radicals. Taurine N-chlorination was determined to demonstrate radiation-induced generation of hypochlorite, a secondary ACS. DNA protection was assessed by determining the extent of DNA fragmentation and plasmid DNA relaxation following exposure to ClO(-) and radiation. Purine base chlorination by ClO(-) and γ-radiation was determined by using 2-aminopurine (2-AP), a fluorescent analog of 6-aminopurine. Chloride anions (Cl(-)) consumed >90% of hydroxyl radicals in physiological solutions produced by γ-radiation resulting in ACS formation, which was detected by (1)H NMR. Importantly, SDG scavenged hypochlorite- and γ-radiation-induced ACS. In addition, SDG blunted ACS-induced fragmentation of calf thymus DNA and plasmid DNA relaxation. SDG treatment before or after ACS exposure decreased the ClO(-) or γ-radiation-induced chlorination of 2-AP. Exposure to γ-radiation resulted in increased taurine chlorination, indicative of ClO(-) generation. NMR studies revealed formation of primary ACS radicals (chlorine atoms (Cl) and dichloro radical anions (Cl2¯)), which were trapped by SDG and its structural analog dopamine. We demonstrate that γ-radiation induces the generation of ACS in physiological solutions. SDG treatment scavenged
Due to the high linear energy transfer and short range of alpha-radiation, targeted radiation therapy using alpha-emitting pharmaceuticals that successfully target small disease clusters will kill target cells with limited harm to healthy tissue, potentially treating the most aggressive forms of cancer. As the parent of a decay chain with four alpha- and two beta-decays, 225Ac is a promising candidate for such a treatment. However, this requires retention of the entire decay chain at the target site, preventing the creation of freely circulating alpha-emitters that reduce therapeutic effect and increase toxicity to non-target tissues. Two major challenges to 225Ac pharmaceutical development exist: insufficient global supply, and the difficulty of preventing toxicity by retaining the entire decay chain at the target site. While TRIUMF works towards large-scale (C i amounts) production of 225Ac, we already use our Isotope Separation On-Line facility to provide small (< 1 mCi) quantities for in-house chemistry and imaging research that aims to improve and assess 225Ac radiopharmaceutical targeting. This presentation provides an overview of this research program and the journey of 225Ac from the beamline to the scanner. This research is funded by the Natural Sciences and Engineering Research Council of Canada.
Aragonite (CaCO3, calcium carbonate) is an abundant biomaterial of marine life. It is the dominant inorganic phase of coral reefs, mollusc bivalve shells and the stalactites or stalagmites of geological sediments. Inorganic and initially precipitate-free aragonite coating solutions (ACS) of pH 7.4 were developed in this study to deposit monolayers of aragonite spherules or ooids on biomaterial (e.g., UHMWPE, ultrahigh molecular weight polyethylene) surfaces soaked in ACS at 30 °C. The ACS solutions of this study have been developed for the surface engineering of synthetic biomaterials. The abiotic ACS solutions, enriched with calcium and bicarbonate ions at different concentrations, essentially mimicked the artificial seawater composition and started to deposit aragonite after a long (4 h) incubation period at the tropical sea surface temperature of 30 °C. While numerous techniques for the solution deposition of calcium hydroxyapatite (Ca10(PO4)6(OH)2), of low thermodynamic solubility, on synthetic biomaterials have been demonstrated, procedures related to the solution-based surface deposition of high solubility aragonite remained uncommon. Monolayers of aragonite ooids deposited at 30 °C on UHMWPE substrates soaked in organic-free ACS solutions were found to possess nano-structures similar to the mortar-and-brick-type botryoids observed in biogenic marine shells. Samples were characterized using SEM, XRD, FTIR, ICP-AES and contact angle goniometry.
Genome-wide experimental studies in Saccharomyces cerevisiae reveal that autonomous replicating sequence (ARS) requires an essential consensus sequence (ACS) for replication activity. Computational studies identified thousands of ACS like patterns in the genome. However, only a few hundreds of these sites act as replicating sites and the rest are considered as dormant or evolving sites. In a bid to understand the sequence makeup of replication sites, a content and context-based analysis was performed on a set of replicating ACS sequences that binds to origin-recognition complex (ORC) denoted as ORC-ACS and non-replicating ACS sequences (nrACS), that are not bound by ORC. In this study, DNA properties such as base composition, correlation, sequence dependent thermodynamic and DNA structural profiles, and their positions have been considered for characterizing ORC-ACS and nrACS. Analysis reveals that ORC-ACS depict marked differences in nucleotide composition and context features in its vicinity compared to nrACS. Interestingly, an A-rich motif was also discovered in ORC-ACS sequences within its nucleosome-free region. Profound changes in the conformational features, such as DNA helical twist, inclination angle and stacking energy between ORC-ACS and nrACS were observed. Distribution of ACS motifs in the non-coding segments points to the locations of ORC-ACS which are found far away from the adjacent gene start position compared to nrACS thereby enabling an accessible environment for ORC-proteins. Our attempt is novel in considering the contextual view of ACS and its flanking region along with nucleosome positioning in the S. cerevisiae genome and may be useful for any computational prediction scheme.
This manuscript describes the development of a new MEMS sensor for the measurement of AC electric current. The sensor is comprised of a MEMS piezoelectric cantilever with a microscale permanent magnet mounted to the cantilever's free end. When placed near a wire carrying AC current, the magnet couples to the oscillating magnetic field surrounding the wire, causing the cantilever to deflect, and piezoelectric coupling produces a sinusoidal voltage proportional to the current in the wire. The sensor is itself passive, requiring no power supply to operate. It also operates on proximity and need only be placed near a current carrier in order to function. The sensor does not need to encircle the current carrier and it therefore can measure current in two-wire zip-cords without necessitating the separation of the two conductors. Applications for tins sensor include measuring residential and commercial electricity use and monitoring electric power distribution networks. An analytical model describing the behavior of the current sensor was developed. This model was also adapted to describe the power output of an energy scavenger coupled to a wire carrying AC current. A mesoscale sensor exhibited a sensitivity of 75 mV/A when measuring AC electric current in a zip-cord. A mesoscale energy scavenger produced 345 muW when coupled to a zip-cord carrying 13 A. MEMS current sensors were fabricated from aluminum nitride piezoelectric cantilevers and composite permanent magnets. The cantilevers were fabricated using a four-mask process. Microscale permanent magnets were dispenser-printed using NdFeB magnetic powder with an epoxy binder. The MEMS AC current sensor was interfaced with amplification circuitry and packaged inside an almninum enclosure. The sensor was also integrated with a mesoscale energy scavenger and power conditioning circuitry to create a fully self-powered current sensor. Unamplified sensitivity of the sensor was 0.1-1.1 mV/A when measuring currents in single 2b1af7f3a8