Selection Of A Novel Aptamer Against Vitronectin Using Capillary Electrophoresis And Next Generation Sequencing

Breast cancer (BC) results in ≃40,000 deaths each year in the United States and even among survivors treatment of the disease may have devastating consequences, including increased risk for heart disease and cognitive impairment resulting from the toxic effects of chemotherapy. Aptamer-mediated drug delivery can contribute to improved treatment outcomes through the selective delivery of chemotherapy to BC cells, provided suitable cancer-specific antigens can be identified. We report here the use of capillary electrophoresis in conjunction with next generation sequencing to develop the first vitronectin (VN) binding aptamer (VBA-01; Kd 405 nmol/l, the first aptamer to vitronectin (VN; Kd = 405 nmol/l), a protein that plays an important role in wound healing and that is present at elevated levels in BC tissue and in the blood of BC patients relative to the corresponding nonmalignant tissues. We used VBA-01 to develop DVBA-01, a dimeric aptamer complex, and conjugated doxorubicin (Dox) to DVBA-01 (7:1 ratio) using pH-sensitive, covalent linkages. Dox conjugation enhanced the thermal stability of the complex (60.2 versus 46.5°C) and did not decrease affinity for the VN target. The resulting DVBA-01-Dox complex displayed increased cytotoxicity to MDA-MB-231 BC cells that were cultured on plasticware coated with VN (1.8 × 10⁻⁶mol/l) relative to uncoated plates (2.4 × 10⁻⁶ mol/l), or plates coated with the related protein fibronectin (2.1 × 10⁻⁶ mol/l). The VBA-01 aptamer was evaluated for binding to human BC tissue using immunohistochemistry and displayed tissue specific binding and apparent association with BC cells. In contrast, a monoclonal antibody that preferentially binds to multimeric VN primarily stained extracellular matrix and vessel walls of BC tissue. Our results indicate a strong potential for using VN-targeting aptamers to improve drug delivery to treat BC

Experimental Signatures of Critically Balanced Turbulence in MAST

Beam Emission Spectroscopy (BES) measurements of ion-scale density fluctuations in the MAST tokamak are used to show that the turbulence correlation time, the drift time associated with ion temperature or density gradients, the particle (ion) streaming time along the magnetic field and the magnetic drift time are consistently comparable, suggesting a "critically balanced" turbulence determined by the local equilibrium. The resulting scalings of the poloidal and radial correlation lengths are derived and tested. The nonlinear time inferred from the density fluctuations is longer than the other times; its ratio to the correlation time scales as $\nu_{*i}^{-0.8\pm0.1}$, where $\nu_{*i}=$ ion collision rate/streaming rate. This is consistent with turbulent decorrelation being controlled by a zonal component, invisible to the BES, with an amplitude exceeding the drift waves' by $\sim \nu_{*i}^{-0.8}$.Comment: 6 pages, 4 figures, submitted to PR

Is public space still possible? : lessons from City Hall Plaza ideas competition

Thesis (M.S.)--Massachusetts Institute of Technology, Dept. of Architecture, 1995.Includes bibliographical references (leaves 101-102).In Autumn, 1994, the City of Boston, in partnership with the Boston Society of Architects and the Building Owners and Managers Association, sponsored the Revitalizing City Hall Plaza Ideas Competition. The Competition sought ideas from design professionals and lay people for ways to popularize and invigorate City Hall Plaza. The program espoused by Mayor Menino was simple: "We must create an atmosphere that will more easily link the public to an open and accessible City Hall, and provide a meeting place for Boston's diverse community." This Competition represents two aspects of thought regarding public space; one specific and one general. Specifically, the effort to revitalize City Hall Plaza is consistent with long-standing desires to create or retain a high level of activity and stability in the Government Center vicinity. In focusing attention on the condition of the existing Plaza, this Competition continues a tradition of seeking a strong anchor for downtown Boston, maintaining an urban vitality and richness, and creating an image bespeaking Boston's regional and national prominence. Generally, the Competition created avenues for discussions of the nature and role of public space. From 190 Competition entries it is possible to determine categories that reflect two different approaches to the design and consideration of public space. One is the Morphological approach, in which considerations of the physical form of the space and the urban fabric are primary. The other is the Programmatic approach, which stresses activities and programmed attractions independently of physical form. Using these entries and categories as data, a comparison of the categories is made which suggests that the Morphological approach is more appropriate for effecting long-term legibility and structure to a city. However, the salient characteristics of the Programmatic cannot be overlooked, especially in a modern economy. Therefore, it is ultimately concluded that the vitality of a city is best served when the immediacy of the Programmatic is appropriately housed in the permanence of the Morphological. To achieve this is to retain a vision of purposes for cities and their spaces beyond mere functionality, which speaks to the aspirations for community and humanness that have historically informed the creation of public space.by Timothy C. Hurley.M.S

The Effect of Moxidectin Treatment on the Equine Hind Gut Microbiome, Metabonome and Feed Fermentation Kinetics in Horses with Very Low Parasite Burdens

Seventeen horses, mean 12 ± 3.5 years, kept at pasture, with haylage provided and no concentrates. Faecal Egg Counts were conducted September 2015‐March 2016, no eggs seen, no anthelmintic given. Sampling commenced March 2016, points were 0 (prior), 16, 48 and 168 hours post anthelmintic. Treatments were randomized, nine animals dosed orally with Moxidectin 18.92 mg/g at 0.4 mg/kg bw and eight controls. Three horses from each group were randomly assigned for fermentation kinetics. Sequencing of the 16S rRNA gene was conducted on extracted faecal bacterial DNA, bioinformatics using QIIME assigning operational taxonomic units (OTUs). LEfSe (Segata et al., 2011) was used to identify differentially abundant OTUs. Bacterial metabolic profiles were characterized by 1H NMR spectroscopy (Escalona et al., 2015), from urine, analysed by Principal Components Analysis. Fermentation of hay and oats, separately, were measured by in vitro gas production (Murray et al., 2006), data were analysed by repeated measures ANOVA

Essential role of fats throughout the lifecycle : summary and recommendations

The document attached has been archived with permission from the editor of the Medical Journal of Australia. An external link to the publisher’s copy is included.Robert A Gibson, Maria Makrides and Coral G B Colye

Experimental and theoretical study of the triple-differential cross section for electron-impact ionization of thymine molecules

Triply differential cross sections for the electron-impact ionization of inner valence orbitals of thymine have been measured using the (e,2e) technique at an incident electron energy of 250 eV. The measurements have been performed with coplanar asymmetric kinematics for scattered electron angles of –10° and –15° and with an ejected-electron energy of 20 eV. Theoretical calculations have been performed within the first-order Born approximation and are in very good agreement with the experimental data.S. M. Bellm, C. J. Colyer, B. Lohmann, and C. Champio

Electron scattering from molecules and molecular aggregates of biological relevance

In this Topical Review we survey the current state of the art in the study of low energy electron collisions with biologically relevant molecules and molecular clusters. We briefly describe the methods and techniques used in the investigation of these processes and summarise the results obtained so far for DNA constituents and their model compounds, amino acids, peptides and other biomolecules. The applications of the data obtained is briefly described as well as future required developments

Real-world diagnostic potential of bacterial biomarkers of canine periodontitis.

INTRODUCTION: The objective of this study was to investigate the diagnostic potential of bacterial biomarkers by comparing the performance of molecular detection assays with clinical assessments of dogs oral health performed by veterinarians. METHODS: Supragingival and subgingival plaque samples were collected from 127 client-owned dogs, pre-booked for procedures under general anesthesia, visiting veterinary practices in the United States. DNA was extracted and bacterial biomarkers quantified using quantitative polymerase chain reaction. Gingivitis and periodontitis were recorded by a trained clinician using the Weighted Gingivitis Periodontitis Score which involved assessing the buccal surfaces of 18 teeth while under general anesthesia. Intraoral dental radiographs of the left and right mandibular first molar teeth were also obtained. These data were then used to establish the diagnostic performance of the molecular assay to detect periodontitis. RESULTS: An initial conscious, visual oral examination performed by the veterinarian identified 67.7% of dogs as having periodontitis, but examination under general anesthesia indicated a higher proportion (86.6%). Analysis of supragingival plaque samples collected by veterinarians from conscious and unconscious dogs demonstrated the assay had an accuracy of 77.7 to 80.9%, a sensitivity of 77.6 to 81.0%, and a specificity of 80.0%. DISCUSSION: Use of this molecular screening tool in conscious dogs has the potential to improve early periodontal disease detection and support veterinary decision making, ultimately improving the oral health of dogs and consequently their quality of life

Elastic electron scattering from the DNA bases cytosine and thymine

Cross-section data for electron scattering from biologically relevant molecules are important for the modeling of energy deposition in living tissue. Relative elastic differential cross sections have been measured for cytosine and thymine using the crossed-beam method. These measurements have been performed for six discrete electron energies between 60 and 500 eV and for detection angles between 15° and 130°. Calculations have been performed via the screen-corrected additivity rule method and are in good agreement with the present experiment.C. J. Colyer, S. M. Bellm, F. Blanco, G. García, and B. Lohman