Conformation of Tax-response elements in the human T-cell leukemia virus type I promoter

Background: HTLV I Tax is believed to activate viral gene expression by binding bZIP proteins (such as CRIB) and increasing their affinities for proviral THE target sites. Each 21 by THE target site contains an imperfect copy of the intrinsically bent CRE target site (the TRE core) surrounded by highly conserved flanking sequences. These flanking sequences are essential for maximal increases in DNA affinity and transactivation, but they are not, apparently, contacted by protein. Here we employ non-denaturing gel electrophoresis to evaluate TRE conformation in the presence and absence of bZIP proteins, and to explore the role of DNA conformation in viral transactivation.Results: Our results show that the TRE-1 flanking sequences modulate the structure and modestly increase the affinity of a CREB bZIP peptide for the TRE-1 core recognition sequence. These flanking sequences are also essential for a maximal increase in stability of the CREB-DNA complex in the presence of Tax.Conclusions: The CRE-like TRE core and the TRE flanking sequences are both essential for formation of stable CRIB-TRE-1 and Tax-CREB-TRE-1 complexes. These two DNA segments may have co-evolved into a unique structure capable of recognizing Tax and a bZIP protein.     

Rapid and flexible synthesis of 1-deoxy-D-xylulose-5-phosphate, the substrate for 1-deoxy-D-xylulose-5-phosphate reductoisomerase

1-Deoxy-D-xylulose-5-phosphate (DXP) is a key intermediate in the non-mevalonate pathway to terpenoids in bacteria, and it is the substrate for the enzyme 1-deoxy-D-xylulose-5-phosphate reductoisomerase (DXP-R). In order to study the mechanism of DXP-R, we required a flexible synthesis of the substrate which would allow the incorporation of isotopic labels, and the variation of the two stereocentres. Thus 1,4-dihydroxypent-2-yne was selectively reduced to give the E-olefin, and selective phosphorylation of the primary alcohol followed by oxidation of the secondary alcohol gave a substrate suitable for dihydroxylation. Dihydroxylation using stoichiometric OsO4 in the presence of chiral ligands gave protected DXP in high ee. Final hydrogenolysis gave DXP in quantitative yield and high purity. DXP-R was produced by rapid cloning of the dxr gene from Escherichia coli through controlled expression and ion exchange chromatography. The synthetic DXP was fully active in enzyme assays catalysed by recombinant DXP-R.     

Efficient NMR enantiodiscrimination of bridge fluorinated paracyclophanes using lanthanide tris β-diketonate complexes

A selection of amino-substituted 1,1,2,2,9,9,10,10 octafluoro[2.2]paracyclophanes were tested for enantiodiscrimination by ¹H and ¹⁹F NMR spectroscopy via their interaction with different lanthanide tris β-diketonate chiral shift reagents. The amino-, and the pseudo-ortho di-amino substituted octafluoro[2.2]paracyclophanes, both of which exhibit planar chirality, revealed significant shifts and splittings of various ¹H and ¹⁹F NMR signals upon the addition of the chiral shift reagents, which allowed the easy determination of the enantiomeric purity. When the chiral shift reagent was added to an inseparable mixture of the (chiral) pseudo-meta, and (achiral) pseudo-para diamino analogues, both the chiral and achiral molecules revealed NMR doubling. In the case of the achiral molecule, this NMR behavior is due to the meso nature of the pseudo-para species.     

Adsorption and Desorption of the Herbicide Thiazafluron as A Function of Soil Properties

Abstract

The effect of soil composition on the adsorption and desorption of the herbicide thiazafluron [1.3-dimethyl-1-(5-trifluoromethyl-1,2,3-thiadiazol-2-yl) urea] by 20 soil samples of 13 selected soil profiles of southern Spain has been studied. The adsorption curves conformed the Freundlich equation and the values of the constants, Kf and nf, ranged from 0.13 to 4.64 and from 0.14 to 1.30, respectively. The simple and multiple regression analysis between Kf and soil properties revealed soil clay content, illite content and CEC as fundamental factors determining thiazafluron adsorption by soils. Unlike other substituted ureas, non significant correlation was found with soil organic matter. Desorption of thiazafluron was hysteretic in all cases, showing and values much lower than those for adsorption. Desorption kinetic indicated that this hysteresis is essentially due to irreversible adsorption, although some degradation seems also to occur. The Freundlich desorption Kfd values were closely related to the same factors as Kf: clay and illite contents and also montmorillonite content, suggesting that most of the hysteresis was due to thiazafluron irreversibly bound to soil clay mineral components.     

A variant of Shelah's characterization of Strong Chang's Conjecture

Shelah considered a certain version of Strong Chang's Conjecture which we denote , and proved that it is equivalent to several statements, including the assertion that Namba forcing is semiproper. We introduce an apparently weaker version, denoted , and prove an analogous characterization of it. In particular, is equivalent to the assertion that the the Friedman‐Krueger poset is semiproper. This strengthens and sharpens results by Cox and sheds some light on problems posed by Usuba, Torres‐Perez and Wu.     

On the expectations of the mathematical knowledge of first-year undergraduates

In a recent survey Sutherland and Dewhurst itemized the extent to which various types of departments expected selected mathematical topics in the background of incoming undergraduates. Using the results of diagnostic tests it is possible to estimate the actual capability of students with given prior qualifications to provide a ‘probable preparedness’ in specific topics. This paper compares expectations with probable preparedness in a range of topics. It is found that while some departments have realistic expectations about the background of their incoming students, others seem over-optimistic about the mathematical knowledge of today's students. Sometimes, topics that the department actually requires must in fact be covered ab initio with the students that they recruit.     

Hybridorubrins A–D: Azaphilone Heterodimers from Stromata of Hypoxylon fragiforme and Insights into the Biosynthetic Machinery for Azaphilone Diversification

The diversity of azaphilones in stromatal extracts of the fungus Hypoxylon fragiforme was investigated and linked to their biosynthetic machineries by using bioinformatics. Nineteen azaphilone-type compounds were isolated and characterized by NMR spectroscopy and mass spectrometry, and their absolute stereoconfigurations were assigned by using Mosher ester analysis and electronic circular dichroism spectroscopy. Four unprecedented bis-azaphilones, named hybridorubrins A–D, were elucidated, in addition to new fragirubrins F and G and various known mitorubrin derivatives. Only the hybridorubrins, which are composed of mitorubrin and fragirubrin moieties, exhibited strong inhibition of Staphylococcus aureus biofilm formation. Analysis of the genome of H. fragiforme revealed the presence of two separate biosynthetic gene clusters (BGCs) hfaza1 and hfaza2 responsible for azaphilone formation. While the hfaza1 BGC likely encodes the assembly of the backbone and addition of fatty acid moieties to yield the (R)-configured series of fragirubrins, the hfaza2 BGC contains the necessary genes to synthesise the widely distributed (S)-mitorubrins. This study is the first example of two distant cross-acting fungal BGCs collaborating to produce two families of azaphilones and bis-azaphilones derived therefrom.     

The effect of music participation on mathematical achievement and overall academic achievement of high school students

A study was conducted on high school students, comparing those with some music credits to those with none. No statistically significant difference was found in their mean math grade point averages (GPA) or their mean cumulative GPAs. Students were then separated into two groups based on the number of music credits. Students who had earned at least two music credits per grade level were placed into Group A. This category included ninth graders with two or more music credits, tenth graders with four or more music credits, eleventh graders with six or more music credits, and twelfth graders with eight or more music credits. The remaining students were placed into Group B. Group A students performed better than group B students. However, the differences were not statistically significant. Scatter plots indicated a slight upward trend in GPAs as the number of music credits increased. Lower GPAs were nonexistent as the music credits increased.