An Intrinsically Fluorescent Recognition Ligand Scaffold Based on Chaperonin Protein and Semiconductor Quantum‐Dot Conjugates

Genetic engineering of a novel protein–nanoparticle hybrid system with great potential for biosensing applications and for patterning of various types of nanoparticles is described. The hybrid system is based on a genetically modified chaperonin protein from the hyperthermophilic archaeon Sulfolobus shibatae. This chaperonin is an 18‐subunit double ring, which self‐assembles in the presence of Mg ions and ATP. Described here is a mutant chaperonin (His‐β‐loopless, HBLL) with increased access to the central cavity and His‐tags on each subunit extending into the central cavity. This mutant binds water‐soluble semiconductor quantum dots, creating a protein‐encapsulated fluorescent nanoparticle. The new bioconjugate has high affinity, in the order of strong antibody–antigen interactions, a one‐to‐one protein–nanoparticle stoichiometry, and high stability. By adding selective binding sites to the solvent‐exposed regions of the chaperonin, this protein–nanoparticle bioconjugate becomes a sensor for specific targets.     

Immobilization of acrylamide-modified oligonucleotides by co-polymerization

A flexible chemistry for solid phase attachment of oligonucleotides is described. Oligonucleotides bearing 5'-terminal acrylamide modifications efficiently co-polymerize with acrylamide monomers to form thermally stable DNA-containing polyacrylamide co-polymers. Co-polymerization attachment is specific for the terminal acrylamide group. Stable probe-containing layers are easily fabricated on supports bearing exposed acrylic groups, including plastic microtiter plates and silanized glass. Attachment can be accomplished using standard polyacrylamide gel recipes and polymerization techniques. Supports having a high surface density of hybridizable oligonucleotide (approximately 200 fmol/mm2) can be produced.     

Molecular requirements for T cell recognition by a major histocompatibility complex class II-restricted T cell receptor: the involvement of the fourth hypervariable loop of the Valpha domain

The role of two central residues (K68, E69) of the fourth hypervariable loop of the Valpha domain (HV4alpha) in antigen recognition by an MHC class II-restricted T cell receptor (TCR) has been analyzed. The TCR recognizes the NH2-terminal peptide of myelin basic protein (Ac1-11, acetylated at NH2 terminus) associated with the class II MHC molecule I-Au. Lysine 68 (K68) and glutamic acid 69 (E69) of HV4alpha have been mutated both individually and simultaneously to alanine (K68A, E69A). The responsiveness of transfectants bearing wild-type and mutated TCRs to Ac1-11-I-Au complexes has been analyzed in the presence and absence of expression of the coreceptor CD4. The data demonstrate that in the absence of CD4 expression, K68 plays a central role in antigen responsiveness. In contrast, the effect of mutating E69 to alanine is less marked. CD4 coexpression can partially compensate for the loss of activity of the K68A mutant transfectants, resulting in responses that, relative to those of the wild-type transfectants, are highly sensitive to anti-CD4 antibody blockade. The observations support models of T cell activation in which both the affinity of the TCR for cognate ligand and the involvement of coreceptors determine the outcome of the T cell-antigen-presenting cell interaction.     

Heat conduction in microbes exposed to pulsed electric fields

Nonthermal pasteurization of liquid foods by intense pulsed electric fields results in advantages over conventional heat pasteurization. Thermal damage to liquid foods can adversely affect the flavor and taste and result in loss of nutrients. Numerical modeling of heating parameters near the microbe during exposure to these intense electric fields is described. Boundary conditions at membrane interfaces included the continuity of temperature and continuity of heat flow. Temperature, heat flow vector, and heat source were included in the one-dimensional model. Two simulations are reported here. Simulation 1 consisted of a 0.5 μm wide microbe suspended in liquid with conductivity of 0.01 S/m and was treated with an applied electric field of 40 kV/cm for 375 ns. Simulation 2 contained a 1.0 μm wide microbe suspended in liquid with conductivity of 0.1 S/m and was treated with an applied electric field of 40 kV/cm for 1 μs. Comparison is made with a uniform conductivity model (ohmic), and it is shown that significant differences exist in the heating parameters between the two models. For the parameters used in these one-dimensional simulations, conditions for electroporation were found to exist without a significant temperature rise in the microbe     

Updated Correlation Between Aircraft Smoke Number and Black Carbon Concentration

Aircraft emissions of black carbon (BC) contribute to anthropogenic climate forcing and degrade air quality. The smoke number (SN) is the current regulatory measure of aircraft particulate matter emissions and quantifies exhaust plume visibility. Several correlations between SN and the exhaust mass concentration of BC (C _BC) have been developed, based on measurements relevant to older aircraft engines. These form the basis of the current standard method used to estimate aircraft BC emissions (First Order Approximation version 3 [FOA3]) for the purposes of environmental impact analyses. In this study, BC with a geometric mean diameter (GMD) of 20, 30, and 60 nm and filter diameters of 19 and 35 mm are used to investigate the effect of particle size and sampling variability on SN measurements. For BC with 20 and 30 nm GMD, corresponding to BC emitted by modern aircraft engines, a smaller SN results from a given C _BC than is the case for BC with 60 nm GMD, which is more typical of older engines. An updated correlation between C _BC and SN that accounts for typical size of BC emitted by modern aircraft is proposed. An uncertainty of ±25% accounts for variation in GMD in the range 20–30 nm and for the range of filter diameters. The SN–C _BC correlation currently used in FOA3 underestimates by a factor of 2.5–3 for SN ≤15, implying that current estimates of aircraft BC emissions derived from SN are underestimated by the same factor.

Copyright 2013 American Association for Aerosol Research     

Effects of iminodipropionitrile on cerebral amino acid levels

Iminodipropionitrile (IDPN), a compound that causes dyskinetic symptoms in animals and has possible use as a model for human dyskinesia, was tested in mice and rats for its effect on cerebral amino acids. In mice, 2 h after IDPN administration, the level of total brain alanine was reduced; after 5 h the levels of aspartic acid and glutamic acid were also reduced, and the level of glutamine was increased. In rats, after chronic administration of IDPN, the level of glutamic acid in the total brain tissue was reduced. After acute administration of IDPN using microdialysis, extracellular GABA and extracellular glutamine levels in the striatum were elevated. This study shows that IDPN causes alterations in total and extracellular levels of neurotransmitter amino acids in the brain, which could have a role in IDPN-induced dyskinesia.     

THE ORIGIN, DISTRIBUTION AND COMPOSITION OF LATERITE

The term laterite means a red rock or red earth deposit. Laterites are formed by the decomposition of different kind of rocks, under conditions yielding aluminum and iron hydroxides. The different theories of origin are discussed, as well as the chemical process of laterization, and the geographic distribution of this peculiar type of clay.