Selective extraction and characterization of a histidine-phosphorylated peptide using immobilized copper(II) ion affinity chromatography and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

Phosphorylation is the predominant posttranslational modification involved in regulating enzymatic activity and mediating signal transduction in prokaryotic and eukaryotic cells. Selective enrichment of phosphorylated peptides prior to mass spectrometric analysis facilitates identification of phosphorylated proteins, determination of specific phosphorylated residues, and characterization of the conditions under which phosphorylation occurs. Such protocols have been established for peptides containing residues that form phosphoesters, such as serine and threonine, using immobilized metal-ion affinity chromatography. Despite the importance of histidine phosphorylation in two-component signal transduction pathways, similar protocols for peptides containing phosphorylated histidine (P-His) residues have proven elusive, due to the instability of these modifications and the propensity of unphosphorylated histidines to interact with immobilized metals ions. We describe a method for the selective extraction of a P-His-containing peptide using immobilized copper(II) ions and disposable metal-chelating pipet tips (ZipTipMC, Millipore). The method is contingent upon pH-dependent interactions between the phosphate group and immobilized copper(II) ions. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry with postsource decay confirms the identity and phosphorylation state of the extracted peptide. Peptides containing unphosphorylated histidine residues or other phosphorylated amino acids are not retained, demonstrating the specificity of the method for P-His-containing peptides.     

Prediction of the multimeric assembly of staphylococcal enterotoxin A with cell-surface protein receptors

Staphylococcal enterotoxin A (SEA) cross-links two class II major histocompatibility complex (MHC) molecules and forms a multimeric assembly with T-cell receptors (TcRs). The X-ray crystal structure of SEA has been solved, yet details describing molecular recognition and association remain unclear. We present a structural model for the interactions of SEA with cell-surface proteins. Molecular docking calculations predicting SEA association with the class II MHC molecule HLA-DR1 were performed by using a rigid-body docking method. Docked orientations were evaluated by a Poisson-Boltzmann model for the electrostatic free energy of binding and the hydrophobic effect calculated from molecular surface areas. We found that the best-scoring SEA conformers for the DR1alpha interface display a binding mode similar to that determined crystallographically for staphylococcal enterotoxin B bound to HLA-DR1. For the zinc-binding site of SEA, docking DR1beta yielded several orientations exhibiting tetrahedral-like coordination geometries. Combining the two interfaces, tetramers were modeled by docking an alphabeta TcR with trimolecular complexes DR1beta-SEA-DR1alpha and SEA-betaDR1alpha-SEA. Our results indicate that the complex DR1beta-SEA-DR1alpha provides a more favorable assembly for the engagement of TcRs, forming SEA molecular contacts that are in accord with reported mutagenesis studies. In contrast, the cooperative association of two SEA molecules on a single DR1 molecule sterically inhibits interactions with TcRs. We suggest that signal transduction stimulated by SEA through large-scale assembly is limited to four or five TcR-(DR1beta-SEA-DR1alpha) tetramers and requires the dimerization of class II MHC molecules, while TcR dimerization is unlikely.     

Management by consent in human-machine systems: when and why it breaks down

This study examined the effects of conflict type, time pressure, and display design on operators' ability to make informed decisions about proposed machine goals and actions in a management-by-consent context. A group of 30 B757 pilots were asked to fly eight descent scenarios while responding to a series of air traffic control clearances. Each scenario presented pilots with a different conflict that arose from either incompatible goals contained in the clearance or inappropriate implementation of the clearance by automated flight deck systems. Pilots were often unable to detect these conflicts, especially under time pressure, and thus failed to disallow or intervene with proposed machine actions. Detection performance was particularly poor for conflicts related to clearance implementation. These conflicts were most likely to be missed when automated systems did more than the pilot expected of them. Performance and verbal protocol data indicate that the observed difficulties can be explained by a combination of poor system feedback and pilots' difficulties with generating expectations of future system behavior. Our results are discussed in terms of their implications for the choice and implementation of automation management strategies in general and, more specifically, with respect to risks involved in envisioned forms of digital air-ground communication in the future aviation system. Actual or potential applications of this research include the design of future data link systems and procedures, as well as the design of future automated systems in any domain that rely on operator consent as a mechanism for human-machine coordination.     

All prompts are created equal,but some prompts are more equal than others

Scores assigned to college placement essays by a computer program (PEG) showed high agreement with the evaluations of human readers (r =.82). Further, both types of graders tended to assign higher or lower scores to essays written about particular topics. Content analyses by a second program (MCCA) indicated that themes in essays varied in terms of emphasis on "analytic," "emotional," or "practical" dimensions. Human and machine readers tended to give higher scores for analytic and practical themes, and lower scores for those involving emotion. The ranks of mean prompt-related grades were concordant with the ranks of mean analytic and practical content across topics. Such findings call for the refined standardization of prompts for future testing, and the need for care in the evaluation of existing essays.     

Shoulder belts in motor vehicle crashes: a statewide analysis of restraint efficacy

The purpose of our study was to evaluate the impact of shoulder belt use on motor vehicle crash ejection, morbidity and mortality. We analyzed motor vehicle crash records linked to hospital inpatient data for front seat occupants of passenger cars in Utah between 1994 and 1996 (n = 103,035). Stochastic simulations were used to adjust for possible seatbelt misclassification. There were 276 (0.3%) occupants coded as using only a shoulder belt. The adjusted odds of ejection for shoulder only belted occupants was higher compared to lap-shoulder belted (odds ratio (OR) = 18.9; 95% confidence interval (CI) = 15.1, 25.1) and lap only belted occupants (OR = 4.3; 95% Cl = 2.9, 7.7). There was no difference in the odds of ejection for an occupant using a shoulder belt only and an occupant using no seatbelt (OR = 1.1; 95% CI = 1.0, 1.3). Occupants using a shoulder belt only were more likely to sustain a fatal or hospitalizing injury than lap-shoulder belted (OR = 2.3; 95% Cl = 1.9, 3.0), and lap only belted occupants (OR = 1.8; 95% CI = 1.3, 2.7), while controlling for other covariates. Occupants using only a shoulder belt had the same odds of a fatal or hospitalizing injury as unbelted occupants (OR = 1.1; 95% Cl = 0.9, 1.4). Average hospital inpatient length of stay, charges and injury severity scores were similar for all restraint types. These results stress the need for the use of a lap belt in conjunction with the shoulder belt.     

Surface chemical characterization of 2.5-microm particulates (PM2.5) from air pollution in Salt Lake City using TOF-SIMS, XPS, and FTIR

Particulate matter with a diameter of 2.5 microm collected in Salt Lake City (SLC PM2.5) was studied using TOF-SIMS (time-of-flight secondary-ion mass spectrometry), XPS (X-ray photoelectron spectroscopy), and FTIR (Fourier transform infrared spectroscopy). The high spatial resolution and high surface sensitivity of TOF-SIMS allow the surfaces of individual particulates to be analyzed. The high mass-resolution of TOF-SIMS provides good separation of signals from different chemical species at the same nominal mass, and the extremely high detection sensitivity of TOF-SIMS makes the detection of trace elements possible. Metallic elements such as Li, Na, Mg, Al, K, Ca, Cr, Mn, Fe, Cu, Zn, Cs, and Bi were detected by TOF-SIMS on the surface of SLC PM25. The uranium ion U+ together with its oxide ions UO+ and UO2+ were also found. Inorganic compounds detected include oxides, hydroxides, nitrates, sulfates, silicates, borates, chlorides, etc. Organic compounds detected include hydrocarbons, alcohols, aldehydes, ethers, carboxylic acids, amines, amides, nitriles, etc. A number of polycyclic aromatic hydrocarbons (PAH) and nitrated polycyclic aromatic hydrocarbons were detected by TOF-SIMS. High-resolution XPS Cls spectrum shows functional groups such as C-O, CO2, C-CO2, C-C, and C-H and aromatic pi-pi* shake-up transitions. High-resolution XPS O 1s spectrum indicates the coexistence of different oxygen compounds on the surface of PM2.5. FTIR results confirm the presence of various organic compounds in SLC PM2.5 detected by TOF-SIMS and XPS.     

Catalytic effects of carbon sorbents for mercury capture

Activated carbon sorbents have the potential to be an effective means of mercury control in combustion systems. Reactions of activated carbons in flow systems with mercury and gas stream components were investigated to determine the types of chemical interactions that occur on the sorbent surface. The effects of carbon type, particle size, temperature, and reactive gases were studied. Sorption kinetics and capacities for lignite- and bituminous-based carbons were compared with those for catalytic carbons at temperatures of 107 degrees C, 150 degrees C, and 163 degrees C. In the air and baseline gas studies, the catalytic carbons exhibited far better sorption than the lignite- and bituminous-derived carbons. With the catalytic carbons, the greater sorption kinetics and capacity in an air stream or baseline gas composition compared with nitrogen provides a clear demonstration that O(2) is required in the gas stream for higher reactivities and capacities. Thus, a catalytic chemisorption mechanism predominates for the sorption of mercury at these conditions. The reaction kinetics are inversely proportional to the temperature, indicating that a preliminary physisorption step with mercury associating with a surface site is rate-determining. In synthetic flue gas streams containing HCl (50 ppm), the sorption kinetics of the catalytic carbon are slightly inferior to those of lignite-based carbon. Thus, the reaction is dominated by a different interaction, where HCl reacts with mercury on the carbon surface and the oxidation sites on the catalytic carbon apparently have no advantage. Granular and fine-particle carbons gave similar results in flue gas streams.     

Band‐Offset Engineering for Enhanced Open‐Circuit Voltage in Polymer–Oxide Hybrid Solar Cells

The power conversion efficiency of organic and hybrid solar cells is commonly reduced by a low open‐circuit voltage (V_OC). In these cases, the V_OC is significantly less than the energy of the lowest energy absorbed photon, divided by the elementary charge q. The low photovoltage originates from characteristically large band offsets between the electron donor and acceptor species. Here a simple method is reported to systematically tune the band offset in a π‐conjugated polymer–metal oxide hybrid donor–acceptor system in order to maximize the V_OC. It is demonstrated that substitution of magnesium into a zinc oxide acceptor (ZnMgO) reduces the band offset and results in a substantial increase in the V_OC of poly(3‐hexylthiophene) (P3HT)–ZnMgO planar devices. The V_OC is seen to increase from 500 mV at x = 0 up to values in excess of 900 mV for x = 0.35. A concomitant increase in overall device efficiency is seen as x is increased from 0 to 0.25, with a maximum power‐conversion efficiency of 0.5 % obtained at x = 0.25, beyond which the efficiency decreases because of increased series resistance in the device. This work provides a new tool for understanding the role of the donor–acceptor band offset in hybrid photovoltaics and for maximizing the photovoltage and power‐conversion efficiency in such devices.     

Human-Body-Model Electrostatic-Discharge and Electrical-Overstress Studies of Buried-Heterostructure Semiconductor Lasers

Optoelectronic components such as laser diodes, light-emitting diodes, and photodiodes are susceptible to electrostatic discharge (ESD) and electrical overstress (EOS). Human-body model (HBM) is the most widely adopted method for the characterization of the ESD performance. In this paper, we report a comprehensive study of the ESD and EOS characteristics of buried-heterostructure (BH) semiconductor lasers using the HBM. Threshold current, optical power, optical spectrum, and reverse-bias current are characterized during the ESD study. We show that the ESD-failure thresholds depend upon the polarity. The chip can sustain the highest ESD stress under forward bias and the lowest one under forward/reverse bias. We also show that the BH lasers exhibit two types of ESD-degradation behavior. The soft degradation is characterized by a gradual increase in the threshold current, whereas the hard degradation is identified by a sudden jump in the threshold current during the ESD voltage ramp. The ESD-degradation behavior seems to be influenced by the cavity length. The failure-analysis results show that about 27% of the ESD failure is related to facet damage. The damage regions occur at the upper laser mesa structure and form preferentially on the bond-pad side. The preferential formation of the facet damage is suggestive of current-crowding effect. We have also found that the ESD-degradation behavior is a function of the facet damage. The soft-degradation failure shows a stronger correlation with the facet damage than the hard-degradation one. Finally, we demonstrate that the ESD performance of the laser can be improved by adding a protection diode.     

Different perspectives: A comparison of newspaper articles to medical examiner data in the reporting of violent deaths

This study compared violent death information reported in state-wide newspaper articles to the medical examiner reports collected for a state public health surveillance system—the National Violent Death Reporting System (NVDRS). While suicides accounted for 83% of deaths in the NVDRS database, more than three-quarters (79%) of violent deaths reported in newspaper articles were homicides. The majority of the suicide incidents were reported in 1-2 newspaper articles whereas the majority of homicide incidents were reported in 11-34 articles. For suicide incidents, the NVDRS reported more circumstances related to mental health problems while newspaper articles reported recent crisis more often. Results show that there is a mismatch in both frequency and type of information reported between a public health surveillance system (NVDRS) and newspaper reporting of violent deaths. As a result of these findings, scientists and other public health professionals may want to engage in media advocacy to provide newspaper reporters with timely and important health information related to the prevention and intervention of violent deaths in their community.