Trypsiligase-Catalyzed Labeling of Proteins on Living Cells

Fluorescent fusion proteins are powerful tools for studying biological processes in living cells, but universal application is limited due to the voluminous size of those tags, which might have an impact on the folding, localization or even the biological function of the target protein. The designed biocatalyst trypsiligase enables site-directed linkage of small-sized fluorescence dyes on the N terminus of integral target proteins located in the outer membrane of living cells through a stable native peptide bond. The function of the approach was tested by using the examples of covalent derivatization of the transmembrane proteins CD147 as well as the EGF receptor, both presented on human HeLa cells. Specific trypsiligase recognition of the site of linkage was mediated by the dipeptide sequence Arg-His added to the proteins’ native N termini, pointing outside the cell membrane. The labeling procedure takes only about 5 minutes, as demonstrated for couplings of the fluorescence dye tetramethyl rhodamine and the affinity label biotin as well.     

Deliberately Losing Control of C−H Activation Processes in the Design of Small-Molecule-Fragment Arrays Targeting Peroxisomal Metabolism

Combined photochemical arylation, “nuisance effect” (S_NAr) reaction sequences have been employed in the design of small arrays for immediate deployment in medium-throughput X-ray protein–ligand structure determination. Reactions were deliberately allowed to run “out of control” in terms of selectivity; for example the ortho-arylation of 2-phenylpyridine gave five products resulting from mono- and bisarylations combined with S_NAr processes. As a result, a number of crystallographic hits against NUDT7, a key peroxisomal CoA ester hydrolase, have been identified.     

Psychotherapy and the twelve step approach for substance abusers: The limits of integration.

Argues that while there is potential for successful blending of 12-step approaches and psychotherapies for chemical dependency at the level of helping strategies/techniques, some basic differences in values and philosophy will make it impossible to ever fully integrate the 2 approaches without compromising one or the other approach. Some of the differences between the 2 approaches involve leadership (indigenous vs professional), individual control (increasing vs decreasing), and spirituality (enhanced vs decreased). Given that psychotherapy and the 12-step approach must remain largely independent, a model for interactions based on mutual respect, independent control, and cooperation is suggested. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Microprocessor-Based Control of an Ion Beam Deposition System

A microprocessor-based system with 32 A/D, 24 D/A, and 16 ac load controllers, has been designed and built to monitor and control an ion beam thin-film deposition system. The A/D and D/A channels have electrical isolation of 7.5 kV between channels and between input and output. The microprocessor system keeps the ion beam deposition parameters stable for extended periods of operation and it is proposed as a means to greatly simplify switching from one deposition species to another to grow thin multilayer or alloy films.     

Object-oriented DBMS requirements

The requirements on an object-oriented DBMS for management of information in a large, complex enterprise are presented. These requirements aid in the achievement of an environment characterized by data sharing, open architectures, application and data portability, and assurance of data integrity. They were defined from the point of view of a user of the DBMS; therefore they describe the expected functionality of the DBMS and do not specify the method of implementation to achieve this functionality. They encompass requirements on the data model, query and data manipulation languages, the system architecure, interfaces to the system, change management, and transaction management.     

Location errors in tachistoscopic recognition: Guesses, probe errors, or spatial confusions?

Examined whether location errors in tachistoscopic visual probe tasks reflect (1) uncertainty about the actual position of items, (2) inappropriate guesses, encouraged by a requirement to respond, or (3) confusion about the position being tested. 46 undergraduates completed probe tasks in which they provided confidence ratings on each trial and tasks that substituted a color cue for a visual pointer or used a double bar probe. Results suggest that location errors on visual probe tasks reflect confusion about the spatial position of target items. Findings are consistent with data questioning the concept of iconic memory and imply that the bar-probe task cannot be used to study precategorical visual memory. (French abstract) (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Modelling the effects of dam removal on migratory walleye (Sander vitreus) early life‐history stages

Many dams in the USA have outlived their intended purpose and an increasing number are being considered for removal. Yet, quantitative studies of the potential physical, biological and ecological responses are needed to assess dam removal decisions. In this paper, the responses of migratory walleye (Sander vitreus) to increased spawning habitat availability as a result of dam removal was studied by comparing scenarios with and without a high‐head dam in the Sandusky River (Ohio), a major tributary to Lake Erie. A conceptual, ecological model was proposed to define the relationship between hydrodynamics and walleye spawning, egg hatching, larval drift and survival. A mathematical, ecological model of the early life‐history stages was then developed and coupled with time series of depth and velocity predictions over the spawning grounds from a 1‐D hydrodynamic model. Model simulations were run for 1984–1993 for both the with‐ and without‐dam scenarios to assess the potential benefit of dam removal. The simulation results demonstrated that velocity, depth and water temperature are major factors influencing adult walleye spawning success. Without the dam, 10 times the amount of spawning habitat would be available for walleye to spawn. This increase in spawning habitat area resulted in up to five times the total egg deposition and seven times the larval output to the nursing grounds, based on the assumption that 5% of the walleye population of Lake Erie migrated up the Sandusky River to spawn. We concluded that the spawning habitat in the current condition (with the dam) is limiting and additional spawning habitat upstream could significantly increase the number of larval walleye drifting to Lake Erie. The model sensitivity analysis showed that the number of walleye migrating up the river in spring is the dominant factor for larval recruitment to the lake. Copyright © 2006 John Wiley & Sons, Ltd.