Identification of genes that are essential to restrict genome duplication to once per cell division.

Nuclear genome duplication is normally restricted to once per cell division, but aberrant events that allow excess DNA replication (EDR) promote genomic instability and aneuploidy, both of which are characteristics of cancer development. Here we provide the first comprehensive identification of genes that are essential to restrict genome duplication to once per cell division. An siRNA library of 21,584 human genes was screened for those that prevent EDR in cancer cells with undetectable chromosomal instability. Candidates were validated by testing multiple siRNAs and chemical inhibitors on both TP53+ and TP53- cells to reveal the relevance of this ubiquitous tumor suppressor to preventing EDR, and in the presence of an apoptosis inhibitor to reveal the full extent of EDR. The results revealed 42 genes that prevented either DNA re-replication or unscheduled endoreplication. All of them participate in one or more of eight cell cycle events. Seventeen of them have not been identified previously in this capacity. Remarkably, 14 of the 42 genes have been shown to prevent aneuploidy in mice. Moreover, suppressing a gene that prevents EDR increased the ability of the chemotherapeutic drug Paclitaxel to induce EDR, suggesting new opportunities for synthetic lethalities in the treatment of human cancers

The involvement of acidic nucleoplasmic DNA-binding protein (and-1) in the regulation of prereplicative complex (pre-RC) assembly in human cells

DNA replication in all eukaryotes starts with the process of loading the replicative helicase MCM2–7 onto chromatin during late mitosis of the cell cycle. MCM2–7 is a key component of the prereplicative complex (pre-RC), which is loaded onto chromatin by the concerted action of origin recognition complex, Cdc6, and Cdt1. Here, we demonstrate that And-1 is assembled onto chromatin in late mitosis and early G(1) phase before the assembly of pre-RC in human cells. And-1 forms complexes with MCM2–7 to facilitate the assembly of MCM2–7 onto chromatin at replication origins in late mitosis and G(1) phase. We also present data to show that depletion of And-1 significantly reduces the interaction between Cdt1 and MCM7 in G(1) phase cells. Thus, human And-1 facilitates loading of the MCM2–7 helicase onto chromatin during the assembly of pre-RC

Long-Range Ordered Carbon Clusters: A Crystalline Material with Amorphous Building Blocks

Solid-state materials can be categorized by their structures into crystalline (having periodic translation symmetry), amorphous (no periodic and orientational symmetry), and quasi-crystalline (having orientational but not periodic translation symmetry) phases. Hybridization of crystalline and amorphous structures at the atomic level has not been experimentally observed. We report the discovery of a long-range ordered material constructed from units of amorphous carbon clusters that was synthesized by compressing solvated fullerenes. Using x-ray diffraction, Raman spectroscopy, and quantum molecular dynamics simulation, we observed that, although carbon-60 cages were crushed and became amorphous, the solvent molecules remained intact, playing a crucial role in maintaining the long-range periodicity. Once formed, the high-pressure phase is quenchable back to ambient conditions and is ultra-incompressible, with the ability to indent diamond

A Comparison of Foliage Profiles in the Sierra National Forest Obtained with a Full-Waveform Under-Canopy EVI Lidar System with the Foliage Profiles Obtained with an Airborne Full-Waveform LVIS Lidar System

Foliage profiles retrieved froma scanning, terrestrial, near-infrared (1064 nm), full-waveformlidar, the Echidna Validation Instrument (EVI), agree well with those obtained from an airborne, near-infrared, full-waveform, large footprint lidar, the Lidar Vegetation Imaging Sensor (LVIS). We conducted trials at 5 plots within a conifer stand at Sierra National Forest in August, 2008. Foliage profiles retrieved from these two lidar systems are closely correlated (e.g., r = 0.987 at 100 mhorizontal distances) at large spatial coverage while they differ significantly at small spatial coverage, indicating the apparent scanning perspective effect on foliage profile retrievals. Alsowe noted the obvious effects of local topography on foliage profile retrievals, particularly on the topmost height retrievals. With a fine spatial resolution and a small beam size, terrestrial lidar systems complement the strengths of the airborne lidars by making a detailed characterization of the crowns from a small field site, and thereby serving as a validation tool and providing localized tuning information for future airborne and spaceborne lidar missions

A Multipronged Comparative Study of the Ultraviolet Photochemistry of 2-, 3-, and 4-Chlorophenol in the Gas Phase

The S1(1ππ*) state of the (dominant) syn-conformer of 2-chlorophenol (2-ClPhOH) in the gas phase has a subpicosecond lifetime, whereas the corresponding S1 states of 3- and 4-ClPhOH have lifetimes that are, respectively, ∼2 and ∼3-orders of magnitude longer. A range of experimental techniques–electronic spectroscopy, ultrafast time-resolved photoion and photoelectron spectroscopies, H Rydberg atom photofragment translational spectroscopy, velocity map imaging, and time-resolved Fourier transform infrared emission spectroscopy–as well as electronic structure calculations (of key regions of the multidimensional ground (S0) state potential energy surface (PES) and selected cuts through the first few excited singlet PESs) have been used in the quest to explain these striking differences in excited state lifetime. The intramolecular O–H···Cl hydrogen bond specific to syn-2-ClPhOH is key. It encourages partial charge transfer and preferential stabilization of the diabatic 1πσ* potential (relative to that of the 1ππ* state) upon stretching the C–Cl bond, with the result that initial C–Cl bond extension on the adiabatic S1 PES offers an essentially barrierless internal conversion pathway via regions of conical intersection with the S0 PES. Intramolecular hydrogen bonding is thus seen to facilitate the type of heterolytic dissociation more typically encountered in solution studies

The fourth phase of the radiative transfer model intercomparison (RAMI) exercise : Actual canopy scenarios and conformity testing

The RAdiative transfer Model Intercomparison (RAMI) activity focuses on the benchmarking of canopy radiative transfer (RT) models. For the current fourth phase of RAMI, six highly realistic virtual plant environments were constructed on the basis of intensive field data collected from (both deciduous and coniferous) forest stands as well as test sites in Europe and South Africa. Twelve RT modelling groups provided simulations of canopy scale (directional and hemispherically integrated) radiative quantities, as well as a series of binary hemispherical photographs acquired from different locations within the virtual canopies. The simulation results showed much greater variance than those recently analysed for the abstract canopy scenarios of RAMI-IV. Canopy complexity is among the most likely drivers behind operator induced errors that gave rise to the discrepancies. Conformity testing was introduced to separate the simulation results into acceptable and non-acceptable contributions. More specifically, a shared risk approach is used to evaluate the compliance of RI model simulations on the basis of reference data generated with the weighted ensemble averaging technique from ISO-13528. However, using concepts from legal metrology, the uncertainty of this reference solution will be shown to prevent a confident assessment of model performance with respect to the selected tolerance intervals. As an alternative, guarded risk decision rules will be presented to account explicitly for the uncertainty associated with the reference and candidate methods. Both guarded acceptance and guarded rejection approaches are used to make confident statements about the acceptance and/or rejection of RT model simulations with respect to the predefined tolerance intervals. (C) 2015 The Authors. Published by Elsevier Inc.Peer reviewe

Evaluation of Cavitation Erosion Behavior of Commercial Steel Grades Used in the Design of Fluid Machinery

The erosion response under cavitation of different steel grades was assessed by studying the erosion rate, the volume removal, the roughness evolution, and the accumulated strain energy. A 20 kHz ltrasonic transducer with a probe diameter of 5 mm and peak-to-peak amplitude of 50 lm was deployed in distilled water to induce damage on the surface of commercial chromium and carbon steel samples. After a relatively short incubation period, cavitation induced the formation of pits, cracks, and craters whose features strongly depended on the hardness and composition of the tested steel. AISI 52100 chromium steel showed the best performance and is, therefore, a promising design candidate for replacing the existing fluid machinery materials that operate within potential cavitating environments

A Directed Molecular Evolution Approach to Improved Immunogenicity of the HIV-1 Envelope Glycoprotein

A prophylactic vaccine is needed to slow the spread of HIV-1 infection. Optimization of the wild-type envelope glycoproteins to create immunogens that can elicit effective neutralizing antibodies is a high priority. Starting with ten genes encoding subtype B HIV-1 gp120 envelope glycoproteins and using in vitro homologous DNA recombination, we created chimeric gp120 variants that were screened for their ability to bind neutralizing monoclonal antibodies. Hundreds of variants were identified with novel antigenic phenotypes that exhibit considerable sequence diversity. Immunization of rabbits with these gp120 variants demonstrated that the majority can induce neutralizing antibodies to HIV-1. One novel variant, called ST-008, induced significantly improved neutralizing antibody responses when assayed against a large panel of primary HIV-1 isolates. Further study of various deletion constructs of ST-008 showed that the enhanced immunogenicity results from a combination of effective DNA priming, an enhanced V3-based response, and an improved response to the constant backbone sequences