Electrochemical Detection of DNA Melting Curves by Means of Heated Biosensors

This article reports about the detection of DNA melting curves at heated electrochemical biosensors. Osmium tetroxide‐bipyridine‐labeled target oligonucleotides are hybridized with probe oligonucleotides immobilized on gold electrodes. Then, the gold electrode is successively heated in order to measure a complete melting curve consisting of alternating current voltammetric signals. Melting temperatures ?_m, determined at various ionic strengths and in dependence on different numbers of base pair mismatches, have been compared with those obtained by means of UV spectrophotometry. The proposed method holds great promise for the fast and easy parallel detection of nucleic acids sequences on selectively heated electrode arrays. A stringent hybridization temperature can be easily adjusted in order to discriminate base pair mismatches.     

Triggered Release of an Active Peptide Conjugate from a DNA Device by an Orally Administrable Small Molecule

A real tonic : In a conceptually new approach to controlled release, the natural daily insulin profile in response to three meals is mimicked (see graph) with release of an insulin conjugate from a matrix, triggered by quinine, a component of tonic water.

     

Detection of Oxidative Damage of Synthetic Oligonucleotides Caused by Thallium(III) Complexes

Biologically important short sequences of DNA were treated with Tl(III) complexes. The chloride complex releases the Tl(III) cation and therefore oxidizes DNA components and at concentrations higher than 50 nM cuts the strands. The cuts are well seen in the EQCM plots. The change in the strand structure is well seen at CD and UV‐Vis spectra. The chelate complexes of Tl(III) do not damage the DNA strands. The SERS spectra were particularly useful in demonstration of the chemical changes and orientation vs. the surface changes of particular groups in the strands after interaction with TlCl₄^?.     

Determination of the Concentrations of Binary Mixtures of Glycerine and Water by a Transient Method

The conductivities of binary mixtures of glycerine and water were measured at 20°C by means of a transient method. The equation describing the correlation between concentration and thermal conductivity was determined. The equation can be used for determining concentrations in mixtures. The results show that (1) the error in the determination of the molar concentration of water in mixtures is less than 1%, (2) the time of measurement is 1 s, (3) this method can be used for on-line analysis in production control. This revised version was published online in July 2006 with corrections to the Cover Date.     

Stabilization of a Bimolecular Triplex by 3′‐S‐Phosphorothiolate Modifications: An NMR and UV Thermal Melting Investigation

Triplexes formed from oligonucleic acids are key to a number of biological processes. They have attracted attention as molecular biology tools and as a result of their relevance in novel therapeutic strategies. The recognition properties of single‐stranded nucleic acids are also relevant in third‐strand binding. Thus, there has been considerable activity in generating such moieties, referred to as triplex forming oligonucleotides (TFOs). Triplexes, composed of Watson–Crick (W–C) base‐paired DNA duplexes and a Hoogsteen base‐paired RNA strand, are reported to be more thermodynamically stable than those in which the third strand is DNA. Consequently, synthetic efforts have been focused on developing TFOs with RNA‐like structural properties. Here, the structural and stability studies of such a TFO, composed of deoxynucleic acids, but with 3′‐S‐phosphorothiolate (3′‐SP) linkages at two sites is described. The modification results in an increase in triplex melting temperature as determined by UV absorption measurements. ¹H NMR analysis and structure generation for the (hairpin) duplex component and the native and modified triplexes revealed that the double helix is not significantly altered by the major groove binding of either TFO. However, the triplex involving the 3′‐SP modifications is more compact. The 3′‐SP modification was previously shown to stabilise G‐quadruplex and i‐motif structures and therefore is now proposed as a generic solution to stabilising multi‐stranded DNA structures.     

Nucleosides and Oligonucleotides with Diynyl Side Chains: Base Pairing and Functionalization of 2′‐Deoxyuridine Derivatives by the Copper(I)‐Catalyzed Alkyne?Azide ‘Click’ Cycloaddition

Oligonucleotides containing the 5‐substituted 2′‐deoxyuridines 1b or 1d bearing side chains with terminal C?C bonds are described, and their duplex stability is compared with oligonucleotides containing the 5‐alkynyl compounds 1a or 1c with only one nonterminal C?C bond in the side chain. For this, 5‐iodo‐2′‐deoxyuridine ( 3 ) and diynes or alkynes were employed as starting materials in the Sonogashira cross‐coupling reaction (Scheme 1). Phosphoramidites 2b – d were prepared (Scheme 3) and used as building blocks in solid‐phase synthesis. T_m Measurements demonstrated that DNA duplexes containing the octa‐1,7‐diynyl side chain or a diprop‐2‐ynyl ether residue, i.e., containing 1b or 1d , are more stable than those containing only one triple bond, i.e., 1a or 1c (Table 3). The diyne‐modified nucleosides were employed in further functionalization reactions by using the protocol of the Cu^I‐catalyzed Huisgen–Meldal–Sharpless [2+3] cycloaddition (‘click chemistry’) (Scheme 2). An aliphatic azide, i. e., 3′‐azido‐3′‐deoxythymidine (AZT; 4 ), as well as the aromatic azido compound 5 were linked to the terminal alkyne group resulting in 1H‐1,2,3‐triazole‐modified derivatives 6 and 7 , respectively (Scheme 2), of which 6 forms a stable duplex DNA (Table 3). The Husigen–Meldal–Sharpless cycloaddition was also performed with oligonucleotides (Schemes 4 and 5).