Cell-Free Synthesis of Selenoproteins in High Yield and Purity for Selective Protein Tagging

The selenol group of selenocysteine is much more nucleophilic than the thiol group of cysteine. Selenocysteine residues in proteins thus offer reactive points for rapid post-translational modification. Herein, we show that selenoproteins can be expressed in high yield and purity by cell-free protein synthesis by global substitution of cysteine by selenocysteine. Complete alkylation of solvent-exposed selenocysteine residues was achieved in 10 minutes with 4-chloromethylene dipicolinic acid (4Cl-MDPA) under conditions that left cysteine residues unchanged even after overnight incubation. Gd^III−Gd^III distances measured by double electron–electron resonance (DEER) experiments of maltose binding protein (MBP) containing two selenocysteine residues tagged with 4Cl-MDPA-Gd^III were indistinguishable from Gd^III−Gd^III distances measured of MBP containing cysteine reacted with 4Br-MDPA tags.     

Patient's self-interpretation of tuberculin skin tests

BACKGROUND: In the outpatient use of tuberculin skin testing (purified protein derivative [PPD]), it is at times inconvenient to have a patient revisit for interpretation. Therefore, we assessed patients' ability to self-interpret these test results. METHODS: In keeping with prior custom, patients were seen by an experienced nurse, who performed skin testing with PPD intermediate strength as well as mumps and Candida anergy control tests in some cases, and explained the procedure. The patients were asked to return 48 to 72 h later, at which time one of the researchers recorded their test interpretations before they were again evaluated by the nurse. RESULTS: Sixty-eight patients were studied, of whom 59 returned at appropriate interval. Eighteen patients had a positive PPD test reaction of 10 to 20 mm induration, which only one patient correctly identified as a positive test result. However, positive anergy control tests were correctly interpreted in 10 of 27 cases. CONCLUSION: The small number of positive PPD test result recognition by these patients may be partially attributed to their lack of education, as well as foreign birth and denial of illness. PPD results should be checked by an experienced professional.     

Creutzfeldt-Jakob disease in a husband and wife

A 53-year-old man died of sporadic Creutzfeldt-Jakob disease (CJD) after a 1.5-year clinical course. Four and a half years later, his then 55-year-old widow died from CJD after a 1-month illness. Both patients had typical clinical and neuropathologic features of the disease, and pathognomonic proteinase-resistant amyloid protein ("prion" protein, or PrP) was present in both brains. Neither patient had a family history of neurologic disease, and molecular genetic analysis of their PrP genes was normal. No medical, surgical, or dietary antecedent of CJD was identified; therefore, we are left with the unanswerable alternatives of human-to-human transmission or the chance occurrence of sporadic CJD in a husband and wife.     

Representation before computation

My main objective is to point out a fundamental weakness in the conventional conception of computation and suggest a promising way out. This weakness is directly related to a gross underestimation of the role of object representation in a computational model, hence confining such models to an unrealistic (input) environment, which, in turn, leads to “unnatural” computational models. This lack of appreciation of the role of structural object representation has been inherited from logic and partly from mathematics, where, in the latter, the centuries-old tradition is to represent objects as unstructured “points”. I also discuss why the appropriate fundamental reorientation in the conception of computational models will bring the resulting study of computation closer to the “natural” computational constrains. An example of the pertinent, class-oriented, representational formalism developed by our group over many years—Evolving Transformation System (ETS)—is briefly outlined here, and several related general lines of research are suggested.     

Thermal explosion in a hot gas mixture with organic gel fuel droplets

Recent experimental evidence has uncovered the peculiar behavior of certain gel droplets which, under appropriate ambient thermal conditions, evaporate and burn in an oscillatory fashion. In this work a preliminary foray is made into the theoretical analysis of the nature of the evolution of a hot gas mixture containing organic gel fuel droplets with oscillatory evaporation within the context of thermal-explosion theory. The problem is modeled as a system of highly nonlinear singularly perturbed ordinary differential equations. Non-dimensionalization of the equations enables identification of the parameters that play a major role in determining the dynamical regimes of the system of equations. The method of integral manifolds is exploited for the analysis and it is found, that for certain parametric regions, the system exhibits new dynamical behavior that is quite different from that found for conventional liquid droplets.     

包容性工程—借助工程技术发明为身体或精神障碍人士赋能

1. Background The use of engineering tools, design, research, and thinking to create environments and capabilities whereby individuals who are currently under-e...     

APOE in non-Alzheimer amyloidoses: transmissible spongiform encephalopathies

We have identified a member of the VEGF family by computer-based homology searching and have designated it VEGF-D. VEGF-D is most closely related to VEGF-C by virtue of the presence of N- and C-terminal extensions that are not found in other VEGF family members. In adult human tissues, VEGF-D mRNA is most abundant in heart, lung, skeletal muscle, colon, and small intestine. Analyses of VEGF-D receptor specificity revealed that VEGF-D is a ligand for both VEGF receptors (VEGFRs) VEGFR-2 (Flk1) and VEGFR-3 (Flt4) and can activate these receptors. However. VEGF-D does not bind to VEGFR-1. Expression of a truncated derivative of VEGF-D demonstrated that the receptor-binding capacities reside in the portion of the molecule that is most closely related in primary structure to other VEGF family members and that corresponds to the mature form of VEGF-C. In addition, VEGF-D is a mitogen for endothelial cells. The structural and functional similarities between VEGF-D and VEGF-C define a subfamily of the VEGFs.     

The glass transition temperature of partially cured polymers as a non-equilibrium parameter and its effect on observed dynamic mechanical behavior

Observations of time dependent behaviors of polymeric systems are often made with the assumption that the glass transition temperature will remain constant during the course of the experiment. This assumption may not be valid for partially cured thermosetting system, where additional cure can increase the glass transition temperature. The observed behavior can be expressed as a time derivative function of two independent variables, the experimental temperature and the state of the system. With proper assumptions, these two variables can be reduced to a single parameter (T ? T_g), whose time derivative will give terms representing the temperature and curing effect separately. Dynamic mechanical measurements have been made on partially cured systems, and the effects of the additional cure on the observed properties are observed. The seemingly complex experimental observations can be qualitatively interpreted with the change of the parameter (T ? T_g).     

Chromatic Dispersion Compensation Using Digital IIR Filtering With Coherent Detection

Digital infinite impulse response (IIR) filtering is proposed as a means for compensating chromatic dispersion in homodyne-detected optical transmission systems with subsequent digital signal processing. Compared to finite impulse response (FIR) filtering, IIR filtering achieves dispersion compensation (DC) using a significantly smaller number of taps. DC of 80 and 160 km in a 10-Gb/s binary phase-shift-keying is experimentally compared for the two filtering schemes. IIR filtering can achieve performance similar to the FIR filtering scheme.