News Article | December 19, 2016
In recent episodes of her nationally syndicated radio program, The Sharon Kleyne Hour Power of Water, Global Climate Change and Your Health on Voice of America, host and water expert Sharon Kleyne called for more responsible water management to avert a deepening world crisis of available fresh water. To that end, Sharon Kleyne urged deeper understanding and more education about the value of water and responsible water management. “Before passage of the Clean Water Act in 1972, Kleyne said, “we were terrible stewards of nature’s gift of water.” Kleyne reminded listeners that we dumped waste and sewage into lakes, rivers, streams and the ocean, damaged natural habitats and endangered future life on the planet. “I’ve always been an advocate of concrete canals, reservoirs and dams,” Kleyne said, “yet we must also pay attention to nature’s lessons, especially when it comes to replenishing water lost to evaporation and run-off.” Sharon Kleyne recommended an essay by Kevin L. Shafer, Executive Director of the Milwaukee Metropolitan Sewerage District. In Healing the Sins of Our Past through Responsible Water Management (collected in the anthology, The Value of Water: A Compendium of Essays by Smart CEOS edited by Donna Vincent Roa & the Value of Water Coalition), and she shared this key passage: “Watersheds are nature’s boundary,” wrote Shafer, “for how water flows over land. As the water flows, it is used and reused. Responsible management of that water means that we will have to track this path and develop integrated approaches from the most upstream point to the most downstream point. The watershed approach will greatly expand the universe of people that must be reached.” Sharon Kleyne pointed out that nature’s infrastructure worked for thousands of years before humans tampered with and messed up so many waterways by creating wasteful run-offs and polluted bodies of water. “Yet, the good news,” said Kleyne, “is that what worked before with water can and will work again.” Kleyne, the founder of Bio-Logic Aqua® Research Water Life Science®, noted that current responsible water management tries to have as little impact on water as possible. “Yes, we need to update water treatment facilities and add new ones,” said Kleyne. “We also need to build more concrete canals to catch and direct rainfall and surface water.” Sharon Kleyne, a cutting edge entrepreneur, who created the first successful global company focusing exclusively on water, is internationally recognized as an educator, research leader and creator of new water technology. She also continues to lead efforts to educate people about the crisis of dehydration due to excess evaporation of earth’s and the body’s water vapor. Kleyne and her company have created a new Water Life Science® lifestyle. To learn more about this lifestyle and the technology behind it, Kleyne encouraged listeners to visit http://www.biologicaqua.com. Sharon Kleyne also said that the natural watershed approach would prove to be cost-effective in the long run, especially. “With nature being allowed once more to do a lot of the work,” Kleyne said, “we’ll save billions of dollars, build better concrete canals, dams and reservoirs, have fewer repairs and reap the benefits of more abundant, cleaner water with new technology. We’ll also save the planet by saving the organisms of all living species including the water, soil, plants humans, animals and the earth’s water vapor that is evaporating at an alarming rate. That’s a concrete water project,” Kleyne concluded, “that I can support!”
News Article | November 30, 2016
Ribosomes from E. coli MRE 600, initiation factors (IF1, IF2, IF3), and fMet-tRNAfMet were prepared as described47, 48, 49. E. coli SelA and SelD (gift from M. Wahl, Free University of Berlin) and SelB50 containing a hexahistidine tag (gift from A. Böck, LMU Munich) were expressed in BL21(DE3) cells and purified according to published protocols4, 51. Biochemical analysis showed that SelB carrying the His-tag is fully functional in the interactions with guanine nucleotides, SECIS elements, and Sec-tRNASec and in mediating UGA recoding in vivo4, 50. tRNASec was overexpressed in E. coli BL21 transformed with the plasmid pCB2013 (gift from A. Böck, LMU Munich)52 and purified and aminoacylated as described4, 53, 54. The mRNA UGA was a derivative of mLP75s54 (Extended Data Fig. 4d). The templates for mRNAs UUC and iSECIS (Extended Data Fig. 4e, f) were prepared by PCR mutagenesis. Transcription and refolding of the mRNAs were performed as described54, 55. 70S ribosomes (3 μM) were incubated with IF1, IF2, IF3 (4.5 μM), mRNA (15 μM), and f[3H]Met-tRNAfMet (7 μM) in buffer A (50 mM Tris-HCl (pH 7.5), 70 mM NH Cl, 30 mM KCl, 7 mM MgCl , 2 mM DTT) with 1 mM GTP for 30 min at 37 °C. Initiation efficiency was close to 100% as verified by nitrocellulose binding. Initiation complexes were purified by gel filtration on a Biosuite 450 HR 5 μm column (Waters). To prepare the ribosome–SelB complexes, ternary complexes SelB–GDPNP–Sec-tRNASec were prepared in buffer B (50 mM Hepes-KOH, pH 7.5, 70 mM NH Cl, 30 mM KCl, 7 mM MgCl , 2 mM DTT) by incubating SelB (1 μM) with GDPNP (2 mM) for 4 min at 37 °C, adding Sec-tRNASec (1 μM) and incubating for 2 min at 23 °C. Ternary complex (0.5 μM) was incubated with initiation complex (0.06 μM) at 0 °C in buffer B supplemented with 0.6 mM spermine and 0.4 mM spermidine before application onto EM grids. Initiation complexes were prepared as described42. Ternary complex SelB–[γ32P]GTP–Sec-tRNASec was prepared in buffer A, by incubating SelB (9 μM) with [γ32P]GTP (60 μM) for 20 min at room temperature (RT), followed by addition of Sec-tRNASec (9 μM) and incubation for 2 min at room temperature. Unbound [γ32P]GTP was removed by gel filtration56. The GTPase activity of SelB was determined at single round conditions, by mixing purified ternary complex (0.05 μM) with UGA, iSECIS and UUC mRNA-programmed initiation complexes at the indicated concentrations. Intrinsic GTP hydrolysis was measured in the absence of ribosomes. When necessary, quench-flow experiments were performed in a KinTek apparatus. Reactions were quenched with formic acid (25% v/v) and the extent of GTP cleavage was determined by thin layer chromatography and phosphor imaging57. The rate of GTP hydrolysis was determined by exponential fitting of the time courses using GraphPad Prism software (GraphPad Software, Inc.); time courses were normalized to the respective reaction end levels. Cryo-EM grids were prepared by applying 5 μl of initiation complex–SelB–GDPNP–Sec-tRNASec complexes onto EM grids (Quantifoil 3.5/1 μm, Jena) covered with pre-floated continuous carbon and subsequently vitrified using a Vitrobot Mark IV (FEI Company, Eindhoven) operated at 4 °C and 100% humidity. 4,000 × 4,000 images (12,681 in total) were acquired in the integration mode by spot-scanning (3 × 3 images per 3.5 μm hole in the Quantifoil carbon film) on a Falcon 2 direct detector (FEI Eindhoven) using a Titan Krios microscope (FEI Eindhoven), fitted with a XFEG electron source (FEI Eindhoven) and a spherical aberration (Cs)-corrector (CEOS Heidelberg). Acquisition was at 300 kV acceleration voltage, an electron dose of ~30 ± 5 electrons per Å2 (determination based on calibration with a Faraday cup), −0.7 to −2.6 μm defocus and a nominal magnification of 118,000× resulting in a final pixel size of ~1.16 Å (determined by optimizing correlation of the final 3D map with atomic coordinates from X-ray crystallography58). The Cs-corrector was tuned as described58 to reduce electron-optical aberrations, linear distortion (to <0.1%) and axial coma (usrimageshift tuning) resulting from the spot-scanning procedure. Ribosome particle images were selected and corrected locally for the contrast-transfer function as described58. Good quality particle images were selected according to power spectra quality (that is, showing Thon rings better than 3.5 Å), yielding a total of 969,526 ribosome particles. An initial 3D structure was computed from randomly selected 100,000 particles to judge the compositional and conformational homogeneity of the image dataset. Even at low-resolution (~10 Å) this average structure showed only very weak, scattered density in the factor-binding region, which disappeared completely upon further refinement to higher-resolution (~3.7 Å) (Extended Data Fig. 1c). This observation indicated that the occupancy of ribosome particles with SelB–Sec-tRNASec was low, in line with the low Sec incorporation efficiency in vivo and in vitro26. The dataset was sorted computationally in a hierarchical manner34 (Extended Data Fig. 1a, b). First, images were sorted according to global ribosome conformation, as described58 (step 1), which allowed us to discard low quality particles and 50S subunits. Subsequently (step 2), particles were sorted according to ligand occupancy using supervised classification by projection matching59 on the basis of a library of ribosome-ligand cryo-EM maps34, 60. Importantly, this library entailed a cryo-EM map of the canonical ribosome–EF-Tu complex60 as potential reference for the ribosome-SelB complex to avoid any reference bias for SelB-Sec-tRNASec, for which no structure was available. To further avoid any high-resolution reference bias, sorting in both steps was performed using low-pass filtered reference maps and particle images binned to about 6.9 Å per pixel. The resulting maps of ribosome-ligand complexes showed all specific features expected for the Sec system (that is, the SECIS in the mRNA, the extra-long variable arm of tRNASec, and domain 4 of SelB). In step 3, the resulting populations of ribosome-ligand particles were further rectified from low-quality particles by three-dimensional (3D) classification in Relion 1.2 and 1.3 (ref. 61). Focused 3D classification in Relion 1.3 without alignment was used to isolate three particle sub-populations that show distinct states of SelB–Sec-tRNASec on the ribosome (step 4); particle images were coarsened to 3.16 Å per pixel for this step to improve the speed and robustness of classification. Finally, to further enrich for ribosome particles of good quality containing SelB within these sub-populations, supervised classification was used in step 5 to assign particles either to the respective ribosome-SelB complex or ribosomes with only peptidyl-site tRNA, but not SelB–Sec-tRNASec bound; again a pixel sampling of 3.16 Å and low-pass filtered reference maps were used to avoid any high-resolution reference bias. In a similar manner, the particle populations of ribosomes carrying only the peptidyl-site tRNA or two tRNAs in the classical state were further computationally cleaned up by supervised classification. The six homogenous ribosome-ligand particle populations resulting from this sorting procedure were refined to high resolution according to the gold-standard procedure in Relion 1.3 and overall resolution of the final maps was determined using high-resolution noise substitution62 in Relion 1.4 (Extended Data Fig. 2a). The final cryo-EM maps were sharpened as described58; and for each reconstruction local, resolution maps were computed using Resmap63 (Extended Data Fig. 2f, g). If not denoted otherwise, densities in the figures are rendered at 2σ, except winged-helix motifs 3–4 and SECIS of the GTPase-activated state, which are rendered at 1.3–1.7σ. Figures were prepared using UCSF Chimera64. Pseudo-crystallographic refinement and model building. For initial model building and refinement, the cryo-EM density map of the GTPase-activated state was sharpened by applying a B factor of −120 Å2, filtered to the estimated highest resolution limit and masked using a pseudo bulk solvent envelope. The mask was obtained by merging the cryo-EM map filtered at different frequencies (9.0 Å, 6.0 Å and 4.0 Å) using the RAVE package65 and UCSF Chimera64. The masked cryo-EM density map was converted to reciprocal space structure factors using Crystallography and NMR System (CNS)66, 67 without employing phase significance blurring scale factors derived from FSC values. Initial rigid body fit of an atomic model of the kirromycin-stalled E. coli ribosome complex58 lacking EF-Tu and Phe-tRNAPhe was performed against the masked cryo-EM map of the GTPase-activated state using Chimera, followed by rigid body refinement of individual chains in PHENIX68 program. The atomic model of SelB with exception of tandem winged-helix motifs 3 and 4 was obtained by combining manual rebuilding and homology modelling with density-guided energy optimization, as implemented in the Rosetta package69, 70, employing a template derived from Methanococcus maripaludis SelB (PDB ID: 4AC9)9 and alignment provided by the HHPRED server71. SelB tandem winged-helix motifs (wh) 1 and 2 were modelled based on the crystal structure of the isolated domain 4 of SelB from Moorella thermoacetica (PDB ID: 1LVA)72; and wh3 and wh4 with mRNA SECIS were modelled on the basis of the crystal structure of E. coli SECIS RNA bound to the domain of elongation factor SelB (PDB ID: 2PJP)73 using Rosetta and Coot74. Sec-tRNASec was manually modelled on the basis of the crystal structure of tRNASec in complex with seryl-tRNA synthetase (PDB ID: 3W3S)6 with Coot and Rcrane75 and optimized by ERRASER76. The catalytic water molecule in the GTPase active centre of SelB was modelled on the basis of the superposition from PDB ID 4V5L35 into available density. Before global optimization of the atomic model of ribosome was performed, models of all individual protein chains were relaxed against the masked cryo-EM map filtered to 3.8 Å resolution (GTPase-activated state) using Rosetta. The assembled complete ribosome model was further minimized in both real and reciprocal space (ERRASER, phenix.real_space_refine77, phenix.refine) with alternating cycles of manual rebuilding in Coot and monitoring the local fit to the density with RESOLVE78. Refinement progress was additionally monitored by calculating the Pearson correlation coefficient (CC ; ref. 79), as well as the Fourier shell correlation between the cryo-EM reconstruction and a model map (FSC ). Both real and reciprocal space refinements in PHENIX employed automatically generated restraints and additional pseudo DEN (Deformable Elastic Network) restraints in order to maintain chemically important interactions. Group atomic displacement factors (ADP) were refined exclusively during reciprocal space refinement steps. During real space refinement steps in phenix.real_space_refine Ramachandran plot restraints were enabled. For parts of the model exhibiting larger conformational differences and/or lower local map resolution, additional cycles of real space refinement and manual fitting were performed against experimental map filtered to lower resolution, which was gradually increased between subsequent refinement steps until convergence. To maintain the intermolecular interactions of selected model fragments within local environment, all residues within at least 15 Å radius were included. For final refinement steps the cryo-EM map was sharpened58 and masked using a smoothed model-based envelope generated within 3.0 Å radius around atoms of the ribosome model of the GTPase-activated state before conversion to reciprocal space structure factors; for these steps and improved visualization the cryo-EM maps resolved at ≤3.6 Å resolution (initial complex, classical and GTPase-activated states) were resampled to a pixel size of 0.789 Å. The final model of the GTPase-activated state consisting of 154,136 individual atoms was refined to 24.90% and 0.936 for R and CC (definition is given below), respectively. The model of the GTPase-activated state exhibits a good stereochemistry with 90.87% of residues in the most favoured region and 0.88% residues in the disallowed region of the Ramachandran plot, protein side chain outliers of 2.83% and all atom clash score 15.09. The remaining states (initial complex, initial binding, codon reading, classical and hybrid states) were modelled based on the final model of GTPase-activated state in an analogous way. For cryo-EM reconstructions resolved at a resolution lower than 3.6 Å, a 3.8 Å radius for generating a smooth model-based envelope was chosen before generating reciprocal space structure factors used for the final refinement steps. FSC and CC , were calculated in a resolution-dependent manner using SFALL program (CCP4 suite80. Real space correlation coefficients (RSCC) were calculated using RESOLVE. All atomic models fit well the experimental cryo-EM map as judged based on the following criteria. (1) The Pearson correlation coefficient (CC ) calculated between model and map structure factors (F and F ) used for refinement is greater than 0.2 for the highest resolution shell and the overall correlation coefficient (CC ) is not lower than 0.9. (2) The calculated FSC value between model map coefficients (F , phase ) and structure factors derived from the cryo-EM map (F , phase ) used for refinement are not lower than 0.5 for the highest resolution shell and overall greater than 0.9. (3) The cumulative RSCC values are greater than 0.8, 0.6 and 0.4 for 53–75%, 88–94% and 95–97% of the residues, respectively. Detailed refinement statistics are presented in Extended Data Table 1, FSC and CC curves in Extended Data Fig. 2a–c. To build the initial recruitment complex, SelB domain 4 from the structure of the isolated SECIS-domain 4 complex (wh2 to wh4; PDB ID: 2PLY)73 was docked onto the initial complex by superimposing the SECIS. Then, wh2 was used to model our structure of SelB–GDPNP–Sec-tRNASec onto the initial complex-domain 4 complex. Refinements of final models against datasets obtained from two half maps were performed at 2.5 Å resolution in PHENIX using several cycles of real space refinement followed by reciprocal space refinement employing reference model restraints. To remove possible model bias a random shift of 0.3 Å was applied to all atomic positions before real space refinement. The FSC and CC were calculated between the models and the masked half-maps used for refinement, as well as between the model and the other half-map for cross-validation. The individual half-maps were masked using a smoothed mask derived from the respective refined model using a radius of 3.0 or 3.8 Å depending on the estimated highest resolution limit. Simulation setup. To obtain the energetics and dynamics of the free ternary complex, molecular dynamics simulations were started from the ribosome-bound conformations (initial binding, codon reading, and GTPase-activated states). Coordinates of Sec-tRNASec, SelB (amino acids 1–401), and GTP were extracted from the cryo-EM structures along with resolved water molecules and ions in the vicinity (<5 Å). The system was protonated, solvated, and ions were added as described earlier81. All simulations were carried out with Gromacs 5 (ref. 82) using the amberff12sb force field83, and the SPC/E water model84. Parameters for potassium and chloride ions were taken from ref. 85 and for modified nucleotides from Aduri et al.86. Atom types for selenocysteine were obtained with ANTECHAMBER87 and partial charges were determined using DFT-B3LYP with a 6–31/G* basis set. Bond and virtual-site constraints, temperature and pressure coupling were applied as described81. For each of the three starting structures, the system was pre-equilibrated as described81 with potential-energy minimization and 50-ns molecular dynamics simulations with position restraints followed by release of position restraints during 20 ns. Production runs started at 70 ns. At times 170, 270, and 370 ns, coordinates were extracted from the trajectory, new velocities were assigned and new simulations were started (12 simulations; total production run simulation time of 24 μs). To address the question of whether intrinsic conformational changes are rate-limiting for the transitions between the free ternary complex and the ribosome-bound states, we carried out a principal component analysis (PCA)88 on three subsets of atoms: ‘v-arm’ (P-atoms of variable arm), ‘no-v-arm’ (remaining peptidyl-atoms of tRNA), and ‘D1’ (Cα-atoms of SelB domain D1). The trajectories were rigid-body fitted using T-stem and acceptor-stem peptidyl-atoms for the ‘v-arm’ and ‘no-v-arm’ subsets and D2 Cα-atoms for the ‘D1’ subsets (Δt = 10 ps, discarding first 200 ns). The trajectories were concatenated and the atomic-displacement covariance-matrix was calculated. The first eigenvectors, sorted according to their eigenvalues, represent the most dominant conformational modes. For ‘no-v-arm’, the first two eigenvectors largely consist of a bending motion of the anticodon arm. To estimate the free-energy landscape of this motion, the projections of all frames onto these eigenvectors were binned. The relative free energy of each bin was calculated via ΔG = −k T ln(c /c ), where k is the Boltzmann constant, T the temperature, c the number of frames in the bin and c is the maximum of all c (Fig. 4c, Extended Data Figs 6a, 7e). For ‘no-v-arm’ conformational modes 1 and 2, the bending angles (α and β) as a function of the projection onto the respective mode were calculated. The vector between the centres of mass of U34–A36 peptidyl-atoms and of T-stem peptidyl-atoms was calculated. For each projection, the angle between the vector and the vector corresponding to the bin with ΔG = 0 was calculated. To monitor the local geometry of the active site in the free ternary complex, the minimal distances between His61 imidazole atoms and GTP atoms (R ) as well as the van-der-Waals distance between Val9 and Met36 (R ) were calculated (Δt = 10 ps, discarding first 200 ns). The resulting free-energy landscape is shown in Extended Data Fig. 7f. To estimate the magnitudes of rates for transitions of tRNASec in the free ternary complex between the conformations corresponding to the ribosome-bound states, the projections onto the “no-v-arm” conformational modes 1 and 2 were calculated, thus obtaining 2-dimensional trajectories. For each simulation the number of transitions n between the region around the free-energy minimum (all bins with 0 ≤ ΔG ≤ 0.5 k T mol−1) and the ribosome-bound conformations was counted. For each ribosome-bound tRNA conformation a region was defined by an ensemble of 10,000 structures generated from the cryo-EM coordinates and b-factors. For each peptidyl-atom and ensemble structure, the coordinate obtained from cryo-EM was shifted in a random direction by distance d drawn from a normal distribution p(d). The normal distribution p(d) with μ = 0 and σ = rmsf (calculated from the b-factor) was set to zero for d < σ and d > σ. The resulting structures were projected onto the ‘no-v-arm’ conformational modes. The region was defined as all bins with entries from these structures. The transition rates were calculated for each simulation by dividing n by the simulation time t . The mean rates and standard deviations were obtained by weighting the individual transition rates by the corresponding t . To address a possible coupling of ‘v-arm’ and ‘no-v-arm’ conformational modes, we investigated whether the projections onto these modes are correlated as would be expected for coupled motions. For all four pairs of eigenvectors v and v , where v is either ‘no-v-arm’ mode 1 or 2 and v is either “v-arm” mode 1 or 2, the correlation coefficient of the projections was calculated. To estimate the statistical error, a bootstrapping method was applied. First, the autocorrelation time τ was defined by f (τ) = exp(−1), where f (Δt) is the autocorrelation function of the projection. For a given pair of eigenvectors v and v , the maximum of the two corresponding τ values (τ ) was used as an interval to resample the projections, resulting in sets of N = t /τ projections. This resampling was repeated 1,000 times and mean and standard deviations of the correlation coefficients were calculated for the simulations started from the individual ribosome-bound conformations and for all simulations combined (Extended Data Fig. 6b, red points). To check whether the resulting correlation coefficients are statistically significant, the standard deviations expected from uncorrelated projections were estimated by randomly drawing N projections (1,000 repetitions; Extended Data Fig. 6b, grey points). To address coupling between tRNA dynamics and motions of SelB domain 1 (D1), the same analysis was carried out for ‘no-v-arm’ and ‘D1’ conformational modes (Extended Data Fig. 6c). Cryo-EM maps/coordinates of the atomic models for each state have been deposited in the Electron Microscopy Data Bank/Protein Data Bank (http://www.emdatabank.org/) with the following accession codes: initial complex, EMD-4121/ 5ZLA; initial binding state, EMD-4122/ 5ZLB; codon reading state, EMD-4123/ 5ZLC; GTPase-activated state, EMD-4124/ 5ZLD; classical state, EMD-4125/ 5ZLE; hybrid state, EMD-4126/ 5ZLF. Cryo-EM micrographs and particle images have been deposited in the EMPIAR database (https://www.ebi.ac.uk/pdbe/emdb/empiar/) with accession code EMPIAR-10077.
News Article | November 30, 2016
Let’s consider the curious legacy of John Sculley. He’s known best for something that happened 31 years ago, when Sculley, then CEO of Apple, sidelined Steve Jobs, which led to Jobs’s exit from the company he founded. Sculley himself left Apple in 1993, when he was fired by the board of directors and replaced by Michael Spindler, better known as "Der Diesel." In almost every history of Apple, including the one Brent Schlender and I wrote, he’s compared unfavorably to Jobs. During a recent 45-minute conversation, I asked Sculley whether it bothered him that people still associate him mostly with something that happened many years ago. "No, not really," he said. "I was at Apple. I worked 10 years there and I learned a lot. I made some mistakes, and we had some great successes. And I’ve been an entrepreneur longer than I ever was CEO of Apple." Even people who seem frozen in time change and grow. Sculley is a great example. "One of my closest friends is Bob Metcalfe," he tells me. Metcalfe, the inventor of Ethernet and founder of the pioneering network company 3Com, hosts what he calls "Big Boys' Camp" at his island in Maine every summer for Sculley and a small group of other friends. "Bob says that every 10 years you have to reinvent yourself. That makes sense to me." After Apple, Sculley helped build MetroPCS, the prepaid cell service that was merged into T-Mobile in 2012. And for the last nine years, he’s been working in the health care industry, trying to bring the consumer sensibility he first developed at Pepsi and honed at Apple to a business that’s regulated, convoluted, illogical and, at times, downright hostile to its customers. Like many others, Sculley believes the U.S. health care system is a mess. But rather than wait for policy makers to straighten it out, Sculley wants to improve things by driving inefficiencies out of the system with companies that combine three things: expertise in information technology and health care, and a consumer-oriented approach to service. The company he’s most focused on now is called RxAdvance, a cloud-based platform that helps pharmaceutical companies, hospitals, and insurers with the management of chronically ill patients living at home. The Affordable Care Act (ACA) exacts severe penalties from insurers whose patients return to the hospital less than a month after being discharged. By improving both the care and the measurement of care at home, RxAdvance can reduce readmissions. The company’s electronic pill box, for example, keeps track of whether a patient has taken his pills as scheduled. RxAdvance can even reduce the number of pills a patient takes by cross-referencing the prescriptions of a patient’s various specialists and doctors. Just before Thanksgiving, Sculley made it clear just how committed he is to RxAdvance by signing on as the company's chairman and chief marketing officer. Other companies Sculley has worked with include a digital prescription manager, an outpatient cardio monitor, a tele-health provider, and a health care data and analytics company. In almost every case, digital technology is improving the connection between patients and providers, a trend that will inevitably survive President-Elect Trump's proposed gutting of the ACA. Sculley’s role is generally that of "a rainmaker," he says. "I know the CEOs of all the major health systems by now, so I spend a lot of time meeting with CEOS, helping to close deals, helping on the consumer story of the business, and mentoring." Sculley says he looks at the health care industry and sees the same kind of opportunity that Jobs saw with the personal computer. "Steve saw that there was a huge potential to sell machines to creative people, and to people who knew nothing about technology, and to make it an incredible experience," he says. "No one else was thinking about that back in the 1980s. And here we are in 2016, and you could say it’s almost the same with people in the health care industry. They are at least a decade behind other industries." That backwardness may portend a market that’s about to explode. "In fintech I can point to 30 companies that have market values between $3 billion and $30 billion. In health tech, there are very few companies with a market value over $1 billion." For Sculley personally, the effort to improve the system is about more than money, and more than restoring his reputation. "I was hanging out at the Mac engineering lab in 1983 with Bill Gates and Steve Jobs," Sculley explains, "and the two of them were talking about their noble cause. They never talked about how much money they were going to make, but it was always about the noble cause: Changing the world one person at a time, giving power to individuals—what Steve called the 'bicycle for the mind.' It’s always stuck with me over the years." Sculley sees the chance to improve health care as his own noble cause. In fact, after nine years, he is so engaged with the problem that he intends to defy Metcalfe’s law of reinvention. "I’m having too much fun," he says. "I won’t necessarily be the individual who will change the world, but this is a clearly solvable problem and an incredible moment."
Alem N.,University of California at Berkeley |
Alem N.,Lawrence Berkeley National Laboratory |
Yazyev O.V.,University of California at Berkeley |
Yazyev O.V.,Lawrence Berkeley National Laboratory |
And 11 more authors.
Physical Review Letters | Year: 2011
Crystalline systems often lower their energy by atom displacements from regular high-symmetry lattice sites. We demonstrate that such symmetry lowering distortions can be visualized by ultrahigh resolution transmission electron microscopy even at single point defects. Experimental investigation of structural distortions at the monovacancy defects in suspended bilayers of hexagonal boron nitride (h-BN) accompanied by first-principles calculations reveals a characteristic charge-induced pm symmetry configuration of boron vacancies. This symmetry breaking is caused by interlayer bond reconstruction across the bilayer h-BN at the negatively charged boron vacancy defects and results in local membrane bending at the defect site. This study confirms that boron vacancies are dominantly present in the h-BN membrane. © 2011 American Physical Society.
Linck M.,CEOS GmbH
Ultramicroscopy | Year: 2013
Amongst the impressive improvements in high-resolution electron microscopy, the Cs-corrector also has significantly enhanced the capabilities of off-axis electron holography. Recently, it has been shown that the signal above noise in the reconstructable phase can be significantly improved by combining holography and hardware aberration correction. Additionally, with a spherical aberration close to zero, the traditional optimum focus for recording high-resolution holograms ("Lichte's defocus") has become less stringent and both, defocus and spherical aberration, can be selected freely within a certain range. This new degree of freedom can be used to improve the signal resolution in the holographically reconstructed object wave locally, e.g. at the atomic positions. A brute force simulation study for an aberration corrected 200. kV TEM is performed to determine optimum values for defocus and spherical aberration for best possible signal to noise in the reconstructed atomic phase signals. Compared to the optimum aberrations for conventional phase contrast imaging (NCSI), which produce "bright atoms" in the image intensity, the resulting optimum values of defocus and spherical aberration for off-axis holography enable "black atom contrast" in the hologram. However, they can significantly enhance the local signal resolution at the atomic positions. At the same time, the benefits of hardware aberration correction for high-resolution off-axis holography are preserved. It turns out that the optimum is depending on the object and its thickness and therefore not universal. © 2012 Elsevier B.V.
Biskupek J.,University of Ulm |
Hartel P.,CEOS GmbH |
Haider M.,CEOS GmbH |
Kaiser U.,University of Ulm
Ultramicroscopy | Year: 2012
The effects of geometric residual aberrations such as coma B 2 and two-fold astigmatism A 1 on the contrast in aberration corrected high resolution transmission electron microscopy (HRTEM) images are investigated on single-walled carbon nanotubes (SWNT). The individual aberrations are adjusted and set up manually using an imaging C S-corrector. We demonstrate how coma B 2 can be recognized by an experienced user directly in the image and how it blurs the contrast. Even with uncorrected (resolution limiting) spherical aberration C S the coma B 2 has to be considered and must be minimized. Limits for a tolerable coma are given. The experiments are confirmed by image simulations. © 2012 Elsevier B.V.
Gibb A.L.,University of California at Berkeley |
Gibb A.L.,Lawrence Berkeley National Laboratory |
Alem N.,University of California at Berkeley |
Alem N.,Lawrence Berkeley National Laboratory |
And 12 more authors.
Journal of the American Chemical Society | Year: 2013
Grain boundaries are observed and characterized in chemical vapor deposition-grown sheets of hexagonal boron nitride (h-BN) via ultra-high-resolution transmission electron microscopy at elevated temperature. Five- and seven-fold defects are readily observed along the grain boundary. Dynamics of strained regions and grain boundary defects are resolved. The defect structures and the resulting out-of-plane warping are consistent with recent theoretical model predictions for grain boundaries in h-BN. © 2013 American Chemical Society.
News Article | September 26, 2016
Here Are the CEOS and Wall Street Titans Trump Will Turn to for Advice A white paper released by the Donald Trump campaign on Monday claims that the Republican presidential nominee’s economic plan, which has been criticized for adding trillions to the national debt while slashing taxes for the wealthy, is nearly revenue neutral, thanks in large part to its proposed overhaul of US trade policy. In the 31-page defense of Trump’s economic plan, University of California-Irvine economist Peter Navarro and private equity investor Wilbur Ross argue that criticism of the GOP nominee’s proposals are unfair because they do not take into account all the positive effects it will have on the economy. “In considering how to score these competing plans fiscally, it is important to note that the Trump plan generates positive and substantial tax revenue offsets from its synergistic suite of trade, regulatory, and energy policy reforms,” they write. “Any analysis that scores the Trump tax cuts in isolation is incomplete and highly misleading.” As it turns out, the effects of Trump’s trade policies are expected to do most of the heavy lifting, adding an estimated $1.74 trillion in additional tax revenue to the US Treasury over the space of 10 years. The problem is that when economists and international trade experts not affiliated with the Trump campaign started reading it, they were not impressed. “Nonsense,” an economist from the Tax Foundation called it. Another from the CATO Institute tweeted out the hashtag #WeAreAllDoomed. A conservative Canadian financial columnist warned a colleague not even to try reading it, saying, “you will poke your eyes out with hot spikes.” Commentators took special exception to Navarro’s and Ross’s discussion of the nature of the country’s Gross Domestic Product and its relationship with the trade deficit. Related: There Are Still Some Big Holes in Trump’s Plan for Economic Growth “The growth in any nation’s gross domestic product (GDP) – and therefore its ability to create jobs and generate additional income and tax revenues – is driven by four factors: consumption growth, the growth in government spending, investment growth, and net exports. When net exports are negative, that is, when a country runs a trade deficit by importing more than it exports, this subtracts from growth.” By simply exporting more and importing less, they argue, US businesses and workers would enjoy much healthier economic growth. However, as experts like economist Alan Cole, pointed out on Monday, the interplay between the elements of GDP is much more complicated than that. Every widget sent overseas is a widget that isn’t consumed domestically. So while sending it overseas helps bring down the negative impact of a trade imbalance, it does so by reducing the impact of domestic consumption on GDP. To be sure, it is possible to increase the productivity of US workers, creating more widgets but there’s precious little evidence in the white paper that suggests that would suddenly occur. In fact, with the Trump plan restricting foreign competition -- as it promises to do -- claiming that US industries would respond with increased efficiency and innovation flies in the face of the laws of economics. “There is so much wrong with it in terms of basic facts and policy,” said Scott Lincicome, a trade attorney and adjunct scholar at the libertarian Cato Institute. Related: Clinton Ups the Ante on the Estate Tax with Her ‘Tax-the-Rich’ Plan Lincicome seemed particularly amazed at a section of the paper in which the authors essentially promise that a Trump administration would play chicken with the World Trade Organization to force a change in Value-Added Tax policy that they believe disadvantages US exporters. (Whether or not it does is very much in question.) The stick that Trump would be wielding in those negotiations would be a US exit from the WTO. “Donald Trump understands that the only way to correct this unfair tax treatment is for the US to use its status as the world’s largest economy, the world’s largest consumer, and the world’s largest importer to put pressure on the WTO to change this unequal treatment,” Navarro and Ross write. “Without the US as a member, there would not be much purpose to the WTO, but prior occupants in the White House have been unwilling to lead on this issue despite its significant negative impacts.” First, according to Lincicome, the idea that Trump could strong arm the WTO – a body where major decisions must be unanimous – is “absurd.” Related: Millions Could Lose Their Health Insurance Under Trump’s Plan to Repeal and Replace Obamacare While Trump might have the authority to pull the US out of the WTO, he could not unilaterally change the country’s existing tariff laws. However, said Lincicome, US withdrawal “would permit every other trading partner in the world to unilaterally block US exports, unilaterally infringe on US intellectual property.” Beyond that, he added, “A US exit from the WTO would just crash markets. The economic uncertainty that would be unleashed by the world’s largest economy leaving a body that has been a pillar of the global economic framework since the 1940s? It would be devastating.” This list of complaints about the Trump proposal from Lincicome and others was long, but the attorney said he managed to find one bright spot. “There is a sliver of good news in this,” he said. “Even right now during campaign season, when politicians are at max hyperbole, we are not seeing any advocacy of the broad-based across the board tariffs that Trump has threatened to do at certain points. “Some have argued that a president Trump actually would have the legal authority to do this kind of stuff,” Lincicome said, though he doubts it himself. At least, he added, “We’re not seeing Trump’s most protectionist trade advisors arguing for that.” In this campaign season, that’s what passes for good news. With Clinton and Trump in a Virtual Tie, Monday’s Debate Could Tip the Balance
News Article | April 18, 2016
On a trip to Spain when he was about 31, Julius Caesar came across a statue of Alexander the Great—and started to cry. This was shortly after the death of Caesar’s wife, many years before he would transform the Roman Empire. A low-level public auditor at the time, Caesar purportedly told his companions, "Do you think I have not just cause to weep, when I consider that Alexander at my age had conquered so many nations, and I have all this time done nothing that is memorable?" I'm 31 myself, and sympathize with Caesar every time I hear a goddamn Justin Bieber song on the radio. It hasn't been long since I realized that many of the writers and musicians and scientists I admire (not to mention Olympic snowboarders) are younger than me. And while I feel more in my element than ever, from time to time I can’t help but hear the voice of the drummer in my college band announce in my head that the good times are over. Perhaps the human zeal for drama explains why magazines print "30 Under 30" issues celebrating youthful overachievers. We admire the fast movers, game changers, and unexpected successes of all stripes. It’s clear to any sports fan that gymnasts and football players peak early. But we’ve got lots of examples of brainy achievers peaking young, too: Albert Einstein and Stephen Hawking developed their most groundbreaking work in physics in their 20s. Alexander Graham Bell invented the telephone at 29. Wolfgang Amadeus Mozart was composing symphonies in his teens. Jesus kicked off a global movement by 33. In a world where actresses over 40 get suddenly swapped for 20-year-old models and Silicon Valley programmers get Botox to avoid being pegged as less innovative, we are perpetually obsessed with perpetual youth—and worse still, in often deeply sexist ways. But are we altogether wrong to peg youth to success? Does our appetite for such stories paint an accurate picture of the relationship between youth and achievement, or just feed into a wrongheaded fantasy? If you’re looking for anecdotes of "past-prime" successes, you’ll find them in almost every category. Peyton Manning just won the Super Bowl at age 40, and Olympians have won medals well into their 50s (and 60s, if you count archery and curling). Quantum physicist Freeman Dyson discovered a groundbreaking solution to the oldest problem in game theory, The Prisoner’s Dilemma, at age 88. So which one is the outlier: Einstein or Manning? F-Dyson or T-Swift? Are 20-year-old actors actually preferable to more seasoned ones? Is a 40-year-old programmer actually going to be less clever on average than a recent grad? Are the good times for a middle-aged working person indeed over, or are they yet to come? I took a look at the data on some of today’s most successful people to find the answers: Data show that (big surprise) athletes indeed tend to peak young, while thinkers like economists and doctors do their best work later in life. In between, we see entertainers and leaders excel in their middle age. These are all awards and "best-of-the-best"-type achievements, so they’re not exactly indicative of what a regular person’s personal best will be. And they also rely to various degrees on public accolades, so the untold numbers of brilliant people who toil in obscurity, and get recognized late or not at all, aren't captured here. Still, it’s interesting to note that from science to art to business to politics, people do tend to reach their apogees during, well, a pretty "average" age for their given fields. Some, like professional athletes and CEOS, tend to cluster especially tightly around certain age ranges (because of constraints like physical prowess and work experience, respectively). However, in each of these fields, people tend to do great work at all sorts of ages. Though Adele pulled the Grammy Album of the Year down from an average of around 40 by winning at age 23, Ray Charles yanked it up by winning his Grammy at 74. To get an ever-better picture of the age striation of people at the top of their games, here’s a look at the ages of the nominees for Best Actor and Best Actress in the Oscars over the last 85 years. After this year especially, the Academy's biases are pretty well-known (I've written more about that here), but we nevertheless see a similarly broad spread. It's true that more women in their 20s have been nominated for Best Actress than women in their 50s, but that data show plenty of actors regardless of gender scoring top honors at every stage of their careers. In his book Old Masters and Young Geniuses, author David Galenson mapped the ages in which "genius" artists in history did their best work and found two patterns: The first he called "experimental innovators," those whose work was exploratory and filled with trial and error and decades of accumulated wisdom and feeling. Those who fit the "experimental" pattern included Virginia Woolf, Jackson Pollock, and Alfred Hitchcock, and they did their best work later in life. Galenson called the other type of artist a "conceptual innovator," or people who break through with radical new ideas. These types of innovation often come from naïveté or recklessness—people who don’t know what they don’t know and therefore inadvertently employ more lateral thinking in their work—and tend to peak at young ages, like Orson Welles, Sylvia Plath, and Pablo Picasso. Notably, renowned artists in both categories produced great work—and high volumes of creative work (good and bad)— throughout their whole lives, as Adam Grant points out in his wonderful new book Originals. The one thing they all truly had in common was that they didn’t quit creating, whether they were young or old, happy or depressed. Personally anyhow, it’s been comforting to see that many of the writers who’ve given a younger me a healthy sense of professional jealousy have addressed this question as well. Malcolm Gladwell has written about late bloomers, and Sam Tanenhaus has explored young writers’ advantage in the New York Times. And they’re both right. The data are clear that we all have the potential to do our best or most creative work at any age. It may be easier to execute a touchdown at 21 and a big-budget film at 51, but that doesn’t mean we can’t or shouldn’t try. In a 2008 article in the Journal of Psychological Science in the Public Interest, researchers charted the range of cognition—or how well our brains can function—at various ages. What we find is that the potential for high-functioning brain activity peaks in our 50s but remains high until our 90s and functional into the 100s; however, the average person peaks at 30 and declines to a nonfunctioning state by his or her late 80s. In other words, while many people do their best work at average ages given what it is that they do, we have the capacity to excel at any age. Truth is, we go through stages in our lives, and there are opportunities to do our best at various periods. What's more, the differences in the way we view accomplishments at different ages can be extremely valuable to society collectively—especially when people work together. The innovation potential at the intersection between "young genius" and "old master," for instance, is huge, especially when both parties are open to listening to and learning from one another. No one wins Best Actress or Actor without the work of a director and a supporting cast. Pulitzer and Hugo winners would go prizeless but for their editors; Grammy winners have producers and songwriters; and Nobel Prizes are often won, to borrow a line from Isaac Newton, by "standing on the shoulders of giants." In other words, the best world is one where we collaborate across ages and disciplines rather than treating life like an Olympic solo event. That’s the realization I had when I re-analyzed the data on U.S. presidents. Neither age nor speed of political climb was correlated with the success as a president. Instead, the best presidents were the most adaptable and open-minded. They were the ones who collaborated best with their cabinets, engaged with their allies and rivals, and used debate and disagreements as fuel for finding the right way to do things, rather than justification for the way they initially wanted to do them. (In fact, this became the basis for the first chapter in my book and this Fast Company story about leadership traits.) But age, it turns out, had nothing to do with it. Julius Caesar’s story with the Alexander the Great statue was all about motivation. He saw someone who had become his best self and decided that he could, too. And Caesar went on to conquer his corner of the world. It didn’t matter that he did it in his 50s and not his 20s. What matters is that he did it.
News Article | February 15, 2017
FILE- In this Feb. 1, 2017, file photo, President Donald Trump, accompanied by his daughter Ivanka, waves as they walk to board Marine One on the South Lawn of the White House in Washington. According to officials, Ivanka Trump, who has been a vocal advocate for policies benefiting working women, was involved in recruiting participants for a round table discussion that will be held Monday, Feb. 13, about women in the workforce. President Donald Trump and Canadian Prime Minister Justin Trudeau will participate in the round table. (AP Photo/Evan Vucci, File) WASHINGTON (AP) -- President Donald Trump and Canadian Prime Minister Justin Trudeau will participate in a roundtable discussion about women in the workforce Monday, showing the rising policy influence of the first daughter who has stressed her commitment to issues like child care. A White House official said the two countries would launch a new task force called the United States Canada Council for the Advancement of Women Business Leaders-Female Entrepreneurs. The official said Trudeau's office reached out to discuss working on a joint effort, noting that this was seen as an area of shared interest between both leaders. Ivanka Trump, who has been a vocal advocate for policies benefiting working women, was involved in recruiting participants and setting the agenda for the meeting and will attend, the official said. Ivanka Trump stressed the importance of maternity leave and child care on the campaign trail, and has recently been meeting with business leaders to discuss those issues. The White House official said that Trump's economic agenda will include a "focus on ensuring women enter and stay in the work force and addressing barriers facing female entrepreneurs." The official requested anonymity to provide details in advance of the meeting. Advancing women has been a clear priority for Trudeau. In late 2015, he drew attention for naming a Cabinet that was 50 percent women, saying that he chose a group that "looks like Canada." Trump did not promise to appoint a gender-balanced Cabinet and has named a smaller number of women and minorities to top jobs. "Our team reached out and suggested as it is an important part of the prime minister's agenda and of our economic growth plan," a Canadian official said. "It seemed like a natural fit given their commitments in their platform as well." The official requested anonymity to discuss the meeting in advance. Trump has offered a childcare plan and has signaled an interest in working on those issues. The business round table will be part of an itinerary that includes a bilateral meeting and a working lunch. The visit is crucial for Canada, which relies heavily on the United States for trade. Trump has said he wants to discuss his plan to overhaul the North American Free Trade Agreement, which involves the United States, Canada and Mexico. There are fears Canada could unintentionally be sideswiped as Trump negotiates with Mexico. Female executives from the United States and Canada are expected for the round table, including General Electric Canada CEO Elyse Allan, TransAlta Corp. CEO Dawn Farrell, Linamar Corp. CEO Linda Hasenfratz, T&T Supermarket Inc. Tina Lee and Schnitzer Steel Industries CEO Tamara Lundgren. Also expected are Julie Sweet, CEO-North America for Accenture, NRStor CEO Annette Verschuren, Monique Leroux, chair of the board of directors for Investissement Québec. Carol Stephenson, of the board of directors for General Motors Co. will attend in place of the GM CEO. Additionally, the meeting will include Katie Telford, Trudeau's chief of staff, Canadian Foreign Minister Chrystia Freeland and Dina Powell, assistant to the president and senior counselor for economic initiatives. Powell, Telford and Freeland were involved in setting up the council and recruiting the CEOS. The council includes many of the meeting attendees, as well as Mary Barra, General Motors CEO, GE Vice Chair Beth Comstock and Catalyst CEO Deborah Gillis. Topics at the event will likely include issues like providing maternity leave and childcare, how to recruit and retain women and how to better support women entrepreneurs. Ivanka Trump does not have an official White House role. But her husband, Jared Kushner, is a senior adviser to the president and she stepped away from her executive positions at the Trump Organization and her lifestyle brand to move her family to Washington. She has been at several public White House events so far and has been privately sitting down with CEOs and thought leaders as she weighs how to pursue her policy interest. Associated Press writer Rob Gillies in Ottawa, Ontario, contributed to this report.