Insertional re-activation of a chloramphenicol acetyltransferase misfolding mutant protein

The deletion of nine residues from the C-terminus of the bacterialchloramphenicol acetyltransferase (CAT) results in depositionof the mutant protein in cytoplasmic inclusion bodies and lossof chloramphenicol resistance in Escherichia coli. This foldingdefect is relieved by C-terminal fusion of the polypeptide withas few as two residues. Based on these observations, efficientpositive selection for the cloning of DNA fragments has beendemonstrated. The cloning vector encodes a C-terminally truncatedCAT protein. Restriction sites in front of the stop codon allowthe insertion of target DNA, resulting in the production ofproperly folded CAT fusion proteins and regained chloramphenicolresistance. The positive selection of recombinants is accomplishedby growth of transformants on chloramphenicol-containing agarplates. The method appears particularly convenient for the cloningof DNA fragments amplified by the PCR because minimal informationto restore CAT folding can be included in the primers. The cloningof random sequences shows that the folding defect can be relievedby fusion to a wide variety of peptides, providing great flexibilityto the positive selection system. This vector may also contributeto the determination of the role of the C-terminus in CAT folding.     

Magnetometer spacing criterion for biomagnetic source currentimaging

A criterion for determining the maximum spacing between magnetometers for measuring the magnetic field is derived. A two-dimensional (2-D) filter model is employed to determine the maximum spatial frequency component present in the magnetic field that is above the spectral noise level. This maximum frequency component is then sampled at a rate greater than twice per period as indicated by the Nyquist criterion, yielding the required magnetometer spacing. It is shown that the rule-of-thumb employed in current clinical biomagnetic array systems, that the spacing between the coils should be approximately equal to the depth of the source, is adequate when the signal-to-noise power ratio is less than 28.4 (14.5 dB). The analysis also quantitatively demonstrates that reducing the separation between the measurement and source planes has a greater effect on the resolution than decreasing the noise level by the same factor. This result is important for employing high T_c superconductor magnetometers that allow thinner thermal insulating layers at the cost of higher thermal noise     

Polymer networks based on α,ω-methacrylate-terminated poly(1,3-dioxolane)

α,ω-Methacrylate-terminated poly(1,3-dioxolane)s (polyDXL) were synthesized by cationic ring-opening polymerization of DXL in the presence of methylene-bis(oxyethylmethacrylate) as transfer agent. If the initiator concentration is small compared with the transfer agent concentration, the molecular weights of the polymers are governed by the ratio of the reacted monomer to the reacted transfer agent. The α,ω-methacrylate-terminated polyDXLs obtained undergo free radical polymerization with formation of polyacetal networks. The properties of the networks as function of the molecular weight of the corresponding prepolymers are reported.     

Low-temperature phase transformation in Li-10 at. % Mg

It is not known whether impaired hematopoiesis noted during human immunodeficiency virus (HIV) infection results from infection of stem/progenitor cells or of cells of the bone marrow microenvironment. Normal adherent primary stromal layers were exposed to HIV to determine which of this mixture of endothelial cells, fibroblasts, and macrophages are susceptible to the virus. Viral p24 in supernatants was noted with monocytotropic HIV-1Ada, HIV-1Ba-L, and HIV-1JR-FL but not with lymphotropic HIV-1LAI nor HIV-1MN strain, and only stromal macrophages expressed the viral antigens. Coculture of the layers with PHA-activated normal lymphocytes failed to rescue lymphotropic virus. No p24 was produced when macrophage-depleted stromal cells were exposed to either HIV-1Ba-L or HIV-1LAI; proviral DNA was then amplified by PCR in cells exposed to either virus, though coculture with lymphocytes rescued only HIV-1Ba-L. Altogether, these data indicate that macrophages are the major targets of HIV in cultured stromal layers. As virus replication in macrophages did not affect the profile of major cytokines involved in regulating hematopoiesis, HIV infection could alter hematopoiesis by other as yet unspecified mechanisms.     

The evolution of custom microarray manufacture

Given the enormous size of the genome and that there are potentially many other types of measurements we need to do to understand it, it has become necessary to pick and choose one's targets to measure because it is still impossible to evaluate the entire genome all at once. What has emerged is a need to have rapidly customizable microarrays. There are two dominant methods to accomplish custom microarray synthesis, Affymetrix-like microarrays manufactured using light projection rather than semiconductor-like masks used by Affymetrix to mass manufacture their GeneChip/sup TM/ arrays now, or the ink-jet printing method employed by Agilent. The manufacture of these custom Affymetrix-like microarrays can now be done on a digital optical chemistry (DOC) machine developed at the University of Texas Southwestern Medical Center, and this method offers much higher feature numbers and feature density than is possible with ink-jet printed arrays. On a microarray, each feature contains a single genetic measurement. The initial DOC prototype has been described in several publications, but that has now led to a second-generation machine. This machine reliably produces a number of arrays daily, has been deployed against a number of biomedical questions, is being used in new ways and has also led to a number of spin-off technologies.     

Functional Properties and Food Applications of Rapeseed Protein Concentrate

Rapeseed protein concentrate (RC), prepared with 2% hexameta-phosphate, was tested for its functionality and performance in some foods. The RC had good nitrogen solubility, fat absorption, emulsification, and whipping capacities but poor water absorption and gelling properties. It increased the emulsion stability, and protein but lowered the fat content of wieners. It also increased the cooking yield, reduced the shrinkage and tenderized meat patties. Results were similar to soybean isolate except for the poorer color and flavor. The cooking yield of RC supplemented wieners was less than the all-meat control and soybean-supplemented wieners. A 9% RC dispersion mixed with an equal volume of eggwhite produced a meringue of comparable stability and texture to that of eggwhite alone.     

Interdisciplinary research: roles for self-organization

No generally accepted principles and guidelines currently exist to help engineers design local interaction mechanisms that result in a desired global behavior. However, several communities have developed ways of approaching this problem in the context of niched application areas. Because the ideas underlying these approaches are often obscured or underemphasized in technical papers, the authors review the role of self-organization in their work. They provide a better picture of the status of the emerging field of self-organizing systems or autonomic computing.     

Systematic characterization of Cl2 reactive ion etchingfor improved ohmics in AlGaN/GaN HEMTs

Pre-metal-deposition reactive ion etching (RIE) was performed on an Al_0.3Ga_0.7N/AlN/GaN heterostructure in order to improve the metal-to-semiconductor contact resistance. An optimum AlGaN thickness for minimizing contact resistance was determined. An initial decrease in contact resistance with etching time was explained in terms of removal of an oxide surface layer and/or by an increase in tunnelling current with the decrease of the AlGaN thickness. The presence of a dissimilar surface layer was confirmed by an initial nonuniform etch depth rate. An increase in contact resistance for deeper etches was experienced. The increase was related to depletion of the two-dimensional (2-D) electron gas (2-DEG) under the ohmics. Etch depths were measured by atomic force microscopy (AFM). The contact resistance decreased from about 0.45 Ωmm for unetched ohmics to a minimum of 0.27 Ωmm for 70 Å etched ohmics. The initial thickness of the AlGaN layer was 250 Å. The decrease in contact resistance, without excessive complications on device processing, supports RIE etching as a viable solution to improve ohmic contact resistance in AlGaN/GaN HEMTs