Spectroscopic characterization of the copper(I)-thiolate cluster in the DNA-binding domain of yeast ACE1 transcription factor

AbstractA polypeptide containing the amino-terminal region of ACEI (residues 1–122; 122*), the activator of yeast Cu-methallothionein gene transcription, shows charge-transfer and metal-centered UV absorption bands, and orange luminescence which are characteristic of Cu-cysteinyl thiolate cluster structures. These spectral features are abolished by the Cu(1) complexing agents CN* and diethyldithiocarbamate or exposure to acid, but not by the Cu(II)chelator. EDTA. Binding of the polypeptide to its specific DNA recognition site, but not to calf-thymus double-stranded DNA, induces quenching of its Tyr and Cu-S cluster luminescence emission. The CD spectrum is characteristic of a tightly folded structure that may be organized around the Cu cluster

Consistent Anisotropic Repulsions for Simple Molecules

We extract atom-atom potentials from the effective spherical potentials that suc cessfully model Hugoniot experiments on molecular fluids, e.g., $O_2$ and $N_2$. In the case of $O_2$ the resulting potentials compare very well with the atom-atom potentials used in studies of solid-state propertie s, while for $N_2$ they are considerably softer at short distances. Ground state (T=0K) and room temperatu re calculations performed with the new $N-N$ potential resolve the previous discrepancy between experimental and theoretical results.Comment: RevTeX, 5 figure

DNA-Interactive Properties of Crotamine, a Cell-Penetrating Polypeptide and a Potential Drug Carrier

Crotamine, a 42-residue polypeptide derived from the venom of the South American rattlesnake Crotalus durissus terrificus, has been shown to be a cell-penetrating protein that targets chromosomes, carries plasmid DNA into cells, and shows specificity for actively proliferating cells. Given this potential role as a nucleic acid-delivery vector, we have studied in detail the binding of crotamine to single- and double-stranded DNAs of different lengths and base compositions over a range of ionic conditions. Agarose gel electrophoresis and ultraviolet spectrophotometry analysis indicate that complexes of crotamine with long-chain DNAs readily aggregate and precipitate at low ionic strength. This aggregation, which may be important for cellular uptake of DNA, becomes less likely with shorter chain length. 25-mer oligonucleotides do not show any evidence of such aggregation, permitting the determination of affinities and size via fluorescence quenching experiments. The polypeptide binds non-cooperatively to DNA, covering about 5 nucleotide residues when it binds to single (ss) or (ds) double stranded molecules. The affinities of the protein for ss-vs. ds-DNA are comparable, and inversely proportional to salt levels. Analysis of the dependence of affinity on [NaCl] indicates that there are a maximum of,3 ionic interactions between the protein and DNA, with some of the binding affinity attributable to non-ionic interactions. Inspection of the three-dimensional structure of the protein suggests that residues 31 to 35, Arg-Trp-Arg-Trp-Lys, could serve as a potential DNA-binding site. A hexapeptide containing this sequence displayed a lower DNA binding affinity and salt dependence as compared to the full-length protein, likely indicative of a more suitable 3D structure and the presence of accessory binding sites in the native crotamine. Taken together, the data presented here describing crotamine-DNA interactions may lend support to the design of more effective nucleic acid drug delivery vehicles which take advantage of crotamine as a carrier with specificity for actively proliferating cells. Citation: Chen P-C, Hayashi MAF, Oliveira EB, Karpel RL (2012) DNA-Interactive Properties of Crotamine, a Cell-Penetrating Polypeptide and a Potential Drug Carrier. PLoS ONE 7(11): e48913. doi:10.1371/journal.pone.0048913University of Maryland Baltimore County Designated Research Initiative Fund, an Undergraduate Research AwardUniversity of Maryland Baltimore County Designated Research Initiative Fund, an Undergraduate Research AwardFundao de Amparo a Pesquisa do Estado de So Paulo [FAPESP]Fundao de Amparo a Pesquisa do Estado de So PauloNational Council of Technological and Scientific Development [CNPq]National Council of Technological and Scientific Developmen

BICEP3: a 95 GHz refracting telescope for degree-scale CMB polarization

BICEP3 is a 550 mm-aperture refracting telescope for polarimetry of radiation in the cosmic microwave background at 95 GHz. It adopts the methodology of BICEP1, BICEP2 and the Keck Array experiments - it possesses sufficient resolution to search for signatures of the inflation-induced cosmic gravitational-wave background while utilizing a compact design for ease of construction and to facilitate the characterization and mitigation of systematics. However, BICEP3 represents a significant breakthrough in per-receiver sensitivity, with a focal plane area 5$\times$ larger than a BICEP2/Keck Array receiver and faster optics ($f/1.6$ vs. $f/2.4$). Large-aperture infrared-reflective metal-mesh filters and infrared-absorptive cold alumina filters and lenses were developed and implemented for its optics. The camera consists of 1280 dual-polarization pixels; each is a pair of orthogonal antenna arrays coupled to transition-edge sensor bolometers and read out by multiplexed SQUIDs. Upon deployment at the South Pole during the 2014-15 season, BICEP3 will have survey speed comparable to Keck Array 150 GHz (2013), and will significantly enhance spectral separation of primordial B-mode power from that of possible galactic dust contamination in the BICEP2 observation patch.Comment: 12 pages, 5 figures. Presented at SPIE Astronomical Telescopes and Instrumentation 2014: Millimeter, Submillimeter, and Far-Infrared Detectors and Instrumentation for Astronomy VII. To be published in Proceedings of SPIE Volume 915

Model Reduction in Flexible-Aircraft Dynamics with Large Rigid-Body Motion

This paper investigates the model reduction, using balanced realizations, of the unsteady aerodynamics of maneuvering flexible aircraft. The aeroelastic response of the vehicle, which may be subject to large wing deformations at trimmed flight, is captured by coupling a displacement-based, flexible-body dynamics formulation with an aerodynamic model based on the unsteady vortex lattice method. Consistent linearization of the aeroelastic problem allows the projection of the structural degrees of freedom on a few vibration modes of the unconstrained vehicle, but preserves all couplings between the rigid and elastic motions and permits the vehicle fiight dynamics to have arbitrarily-large angular velocities. The high-order aerodynamic system, which defines the mapping between the small number of generalized coordinates and unsteady aerodynamic loads, is then reduced using the balanced truncation method. Numerical studies on a representative high-altitude, long-endurance aircraft show a very substantial reduction in model size, by up to three orders of magnitude, that leads to model orders (and computational cost) similar to those in conventional frequency-based methods but with higher modeling fidelity to compute maneuver loads. Closed-loop results for the Goland wing finally demonstrate the application of this approach in the synthesis of a robust flutter suppression controller. © 2013 by Henrik Hesse and Rafael Palacios

BICEP2 / Keck Array VIII: Measurement of gravitational lensing from large-scale B-mode polarization

We present measurements of polarization lensing using the 150 GHz maps which include all data taken by the BICEP2 & Keck Array CMB polarization experiments up to and including the 2014 observing season (BK14). Despite their modest angular resolution ($\sim 0.5^\circ$), the excellent sensitivity ($\sim 3\mu$K-arcmin) of these maps makes it possible to directly reconstruct the lensing potential using only information at larger angular scales ($\ell\leq 700$). From the auto-spectrum of the reconstructed potential we measure an amplitude of the spectrum to be $A^{\phi\phi}_{\rm L}=1.15\pm 0.36$ (Planck $\Lambda$CDM prediction corresponds to $A^{\phi\phi}_{\rm L}=1$), and reject the no-lensing hypothesis at 5.8$\sigma$, which is the highest significance achieved to date using an EB lensing estimator. Taking the cross-spectrum of the reconstructed potential with the Planck 2015 lensing map yields $A^{\phi\phi}_{\rm L}=1.13\pm 0.20$. These direct measurements of $A^{\phi\phi}_{\rm L}$ are consistent with the $\Lambda$CDM cosmology, and with that derived from the previously reported BK14 B-mode auto-spectrum ($A^{\rm BB}_{\rm L}=1.20\pm 0.17$). We perform a series of null tests and consistency checks to show that these results are robust against systematics and are insensitive to analysis choices. These results unambiguously demonstrate that the B-modes previously reported by BICEP / Keck at intermediate angular scales ($150\lesssim\ell\lesssim 350$) are dominated by gravitational lensing. The good agreement between the lensing amplitudes obtained from the lensing reconstruction and B-mode spectrum starts to place constraints on any alternative cosmological sources of B-modes at these angular scales.Comment: 12 pages, 8 figure

Unraveling the antifungal activity of a South American rattlesnake toxin crotamine

Crotamine is a highly basic peptide from the venom of Crotalus durissus terrificus rattlesnake. Its common gene ancestry and structural similarity with the beta-defensins, mainly due to an identical disulfide bond pattern, stimulated us to assess the antimicrobial properties of native, recombinant, and chemically synthesized crotamine. Antimicrobial activities against standard strains and clinical isolates were analyzed by the colorimetric microdilution method showing a weak antibacterial activity against both Gram-positive and Gram-negative bacteria [MIC (Minimum Inhibitory Concentration) of 50->200 mu g/mL], with the exception of Micrococcus luteus [MIC ranging from 1 to 2 mu g/mL]. No detectable activity was observed for the filamentous fungus Aspergillus fumigatus and Trichophyton rubrum at concentrations up to 125 mu g/mL. However, a pronounced antifungal activity against Candida spp., Trichosporon spp., and Cryptococcus neoformans [12.5-50.0 mu g/mL] was observed. Chemically produced synthetic crotamine in general displayed MIC values similar to those observed for native crotamine, whereas recombinant crotamine was overridingly more potent in most assays. On the other hand, derived short linear peptides were not very effective apart from a few exceptions. Pronounced ultrastructure alteration in Candida albicans elicited by crotamine was observed by electron microscopy analyses. the peculiar specificity for highly proliferating cells was confirmed here showing potential low cytotoxic effect of crotamine against nontumoral mammal cell lines (HEK293, PC12, and primary culture astrocyte cells) compared to tumoral B16F10 cells, and no hemolytic activity was observed. Taken together these results suggest that, at low concentration, crotamine is a potentially valuable anti-yeast or candicidal agent, with low harmful effects on normal mammal cells, justifying further studies on its mechanisms of action aiming medical and industrial applications. (C) 2012 Elsevier Masson SAS. All rights reserved.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)University of Maryland Baltimore County Designated Research Initiative FundUniversidade Federal de São Paulo UNIFESP, Dept Farmacol, BR-04044020 São Paulo, BrazilUniversidade Federal de São Paulo UNIFESP, Dept Med, BR-04044020 São Paulo, BrazilUniv São Paulo, Dept Bioquim & Imunol, BR-14049 Ribeirao Preto, BrazilUniv Maryland, Sch Med, Inst Human Virol, Baltimore, MD 21201 USAUniv Maryland, Dept Biochem & Mol Biol, Baltimore, MD 21201 USAUniversidade Federal de São Paulo UNIFESP, Dept Bioquim, BR-04044020 São Paulo, BrazilInst Butantan, Lab Bioquim & Biofis, BR-05503900 São Paulo, BrazilUniversidade Federal de São Paulo UNIFESP, Dept Ginecol, BR-04044020 São Paulo, BrazilCBA, Lab Bioquim & Biol Mol, Manaus, Amazonas, BrazilUniv Maryland Baltimore Cty, Dept Chem & Biochem, Baltimore, MD 21250 USAInst Butantan, Ctr Toxinol Aplicada CAT CEPID, BR-05503900 São Paulo, BrazilUniversidade Federal de São Paulo UNIFESP, Dept Farmacol, BR-04044020 São Paulo, BrazilUniversidade Federal de São Paulo UNIFESP, Dept Med, BR-04044020 São Paulo, BrazilUniversidade Federal de São Paulo UNIFESP, Dept Bioquim, BR-04044020 São Paulo, BrazilUniversidade Federal de São Paulo UNIFESP, Dept Ginecol, BR-04044020 São Paulo, BrazilWeb of Scienc