Coffee brewing sonoreactor for reducing the time of cold brew from several hours to minutes while maintaining sensory attributes.

This research designed and developed an ultrasonic reactor for a fast and on demand production of cold brew coffee, remarkably reducing the brewing time from 24 h to less than 3 min. The technology was engineered by utilizing resonance to induce ultrasonic waves around the walls of the brewing basket of an espresso machine. The sound transmission system comprised a transducer, a horn and a brewing basket. This arrangement transformed the coffee basket into an effective sonoreactor that injected sound waves at multiple points through its walls, thereby generating multiple regions for acoustic cavitation within the reactor. Furthermore, acoustic streaming induced greater mixing and enhanced mass transfer during brewing. The design was accomplished by modeling the transmission of sound, and acoustic cavitation. Brew characterization and chemical composition analysis was performed, considering factors such as pH, acidity, color, and the composition of caffeine, fatty acids, and volatiles. The efficiency of the extraction increased by decreasing the basket loading percentage (BLP). For instance, sonicating at 100 W doubled the extraction yield and caffeine concentration, from 15.05 % to 33.44 % at BLP = 33 %, and from 0.91 mg/mL to 1.84 mg/mL at BLP = 67 %, respectively. The total fatty acids increased from 1.16 mg/mL to 9.20 mg/mL, representing an eightfold increase, at BLP = 33 %. Finally, a sensory analysis was conducted to evaluate appearance, aroma, texture, flavor, and aftertaste, which demonstrated that coffee brewed for 1 and 3 min in the sonoreactor exhibited almost undistinguishable properties compared to a standard 24 h brewing without ultrasound.

Fast and Sensitive Detection of Ammonia from Electrochemical Nitrogen Reduction Reactions by 1 H NMR with Radiation Damping.

A facile NMR method is reported for analysis of ammonia from the electrochemical reduction of nitrogen, which compares a calibrated colorimetric method, a calibrated 1 H NMR method and two 1 H NMR direct measurements using external reference materials. Unlike spectrophotometric methods, 1 H NMR requires less bench time and does not require separation of ammonia from the electrolyte. A novel approach to the problem of radiation damping in NMR measurements considered the specific role of hardware tuning. Radiation damping is suppressed improving signal-to-noise ratio and detection limit (1.5 µg L-1 ). The method is demonstrated to be effective for the analysis of ammonia from direct electrochemical nitrogen reduction in KOH, and from lithium-mediated nitrogen reduction in a non-aqueous solution. An uncertainty budget is prepared for the measurement of ammonia.

AMP-activated protein kinase activators have compound and concentration-specific effects on brain metabolism.

AMP-activated protein kinase (AMPK) is a key sensor of energy balance playing important roles in the balancing of anabolic and catabolic activities. The high energy demands of the brain and its limited capacity to store energy indicate that AMPK may play a significant role in brain metabolism. Here, we activated AMPK in guinea pig cortical tissue slices, both directly with A769662 and PF 06409577 and indirectly with AICAR and metformin. We studied the resultant metabolism of [1-13 C]glucose and [1,2-13 C]acetate using NMR spectroscopy. We found distinct activator concentration-dependent effects on metabolism, which ranged from decreased metabolic pool sizes at EC50 activator concentrations with no expected stimulation in glycolytic flux to increased aerobic glycolysis and decreased pyruvate metabolism with certain activators. Further, activation with direct versus indirect activators produced distinct metabolic outcomes at both low (EC50 ) and higher (EC50 × 10) concentrations. Specific direct activation of ß1-containing AMPK isoforms with PF 06409577 resulted in increased Krebs cycle activity, restoring pyruvate metabolism while A769662 increased lactate and alanine production, as well as labelling of citrate and glutamine. These results reveal a complex metabolic response to AMPK activators in brain beyond increased aerobic glycolysis and indicate that further research is warranted into their concentration- and mechanism-dependent impact.

Molecular Recognition Patterns between Vitamin B12 and Proteins Explored through STD-NMR and In Silico Studies.

Ligand-receptor molecular recognition is the basis of biological processes. The Saturation Transfer Difference-NMR (STD-NMR) technique has been recently used to gain qualitative and quantitative information about physiological interactions at an atomic resolution. The molecular recognition patterns between the cyanocobalamin (CNBL)/aqua cobalamin (OHBL) and different plant and animal proteins were investigated via STD-NMR supplemented by molecular docking. This study demonstrates that myoglobin has the highest binding affinity and that gluten has the lowest affinity. Casein also shows a higher binding affinity for cyanocobalamin when compared with that of plant-based proteins. STD-NMR results showed the moderate binding capability of casein with both CNBL and OHBL. Computer simulation confirmed the recognition mode in theory and was compared with the experiments. This work is beneficial for understanding the binding affinity and biological action of cyanocobalamin and will attract researchers to use NMR technology to link the chemical and physiological properties of nutrients.

Molecular dynamic simulations of diacridine binding to DNA: Indications that C6 diacridine can bisintercalate spanning two base pairs.

Dimers of 9-aminoacridine linked via the 9-amino group with polymethylene chains, termed diacridines, are known to bisintercalate into DNA when the linker comprises 6 or more methylene units. There are no literature reports of crystal or NMR solution structures for bisintercalated diacridine-DNA complexes, and the issue of the structure of the C6 ([CH2 ]n linker where n = 6) diacridine complex remains unresolved. Previously, based on simple geometric considerations, it was proposed that C6 diacridine could only span a single base pair, which requires that its bifunctional reaction violates the widely-observed "neighbor exclusion principle" where bound intercalators are separated by at least 2 base pairs. Here we have explored the structure of diacridine-DNA complexes using unrestrained molecular dynamics in explicit solvent using the parmbsc0 forcefield in AMBER14. We studied the C4 to C8 dimers, intercalated via both the minor and major DNA grooves, to a variety of nucleotide sequences. We find that C6, C7, and C8 diacridine are able to form 2 base pair bisintercalated complexes from either groove, whereas the C4 and C5 homologues cannot. We conclude that C6 diacridine does have the capacity to bisintercalate without violating neighbor exclusion, and that the previous proposed binding model needs revision.

Structures and dynamics of DNA complexes of the desmethyl analog of the cytotoxin MLN944: Insights into activity when a methyl isn't futile.

Structure activity relationships for tricyclic-carboxamide topoisomerase II poisons indicate that cytotoxicity is enhanced by the presence of methyl, and other, groups in the position peri to the carboxamide. Linked dimers of phenazine-1-carboxamides are potent cytotoxins and one phenazine dimer, MLN944 (alternatively XR5944), has been in clinical trial. MLN944 is a template inhibitor of transcription, whereas corresponding monomers are not. Nevertheless, its cytotoxic potency is also diminished by removal of its peri methyl groups. Here, we describe NMR and molecular dynamic studies of the interaction of desmethyl MLN944 with d(ATGCAT)2 , d(TATGCATA)2 , and d(TACGCGTA)2 to investigate the influence of the nine-methyl group on the structure of MLN944 complexes. As with MLN944, the carboxamide group hydrogen bonds to the phenazine ring nitrogen, the ligand sandwiches the central GC base pairs in the major groove, and the protonated linker amines hydrogen bond primarily to the O6 atom of the guanines. Molecular dynamics studies reveal that the linker exists in multiple conformations, none of which produce an ideal set of hydrogen bonds. In distinction, however, the carboxamide-to-phenazine ring nitrogen hydrogen bond is weaker, the overall helix winding is less and the NMR resonances are broader in the desmethyl complexes. Exchange between free and complexed DNA, quantified using two-dimensional NOESY spectra, is faster for the desmethyl MLN944 complexes than for MLN944 complexes. Overall, the data suggest that desmethyl MLN944 DNA complexes are "looser" and more unwound at the binding site, leading to faster dissociation rates, which could account for the diminished efficacy of the desmethyl analog.

Biochar-based fertilizer: Supercharging root membrane potential and biomass yield of rice.

Biochar-based compound fertilizers (BCF) and amendments have proven to enhance crop yields and modify soil properties (pH, nutrients, organic matter, structure etc.) and are now in commercial production in China. While there is a good understanding of the changes in soil properties following biochar addition, the interactions within the rhizosphere remain largely unstudied, with benefits to yield observed beyond the changes in soil properties alone. We investigated the rhizosphere interactions following the addition of an activated wheat straw BCF at an application rates of 0.25% (g·g-1 soil), which could potentially explain the increase of plant biomass (by 67%), herbage N (by 40%) and P (by 46%) uptake in the rice plants grown in the BCF-treated soil, compared to the rice plants grown in the soil with conventional fertilizer alone. Examination of the roots revealed that micron and submicron-sized biochar were embedded in the plaque layer. BCF increased soil Eh by 85 mV and increased the potential difference between the rhizosphere soil and the root membrane by 65 mV. This increased potential difference lowered the free energy required for root nutrient accumulation, potentially explaining greater plant nutrient content and biomass. We also demonstrate an increased abundance of plant-growth promoting bacteria and fungi in the rhizosphere. We suggest that the redox properties of the biochar cause major changes in electron status of rhizosphere soils that drive the observed agronomic benefits.

Brewing coffee? - Ultra-sonication has clear beneficial effects on the extraction of key volatile aroma components and triglycerides.

Ultrasound has been investigated as a new technique for brewing coffee. A two-level factorial experimental design was conducted to identify the effects of ultra-sonication on the extraction of coffee components during ultrasonically-assisted coffee brewing. Different brews were produced by aqueous extraction from roasted ground coffee beans with sonication, and without it as a control, by varying coffee concentration (5% and 10% w/w), temperature (25 and 50 °C) and sonication time (1 and 5 min). These brews were tested for antioxidant capacity (using the ABTS assay), caffeine and triglycerides (using quantitative NMR spectroscopy) and specific aroma/flavour volatiles (using headspace SPME-GC-MS). Additional observations of colour, foaming, body and flavour were also reported. Ultrasound was found to significantly increase the extraction of caffeine, triglycerides and several of the key volatile compounds from coffee, although it did appear to decrease the concentration of antioxidants over the controls, especially with longer time and higher temperature. Furthermore, all the sonicated samples exhibited a lighter caramel colour and lower foam formation which were attributed to their higher triglyceride content. The increased concentration of triglycerides and volatiles were by far the most outstanding responses.

Application of low-field, 1H/13C high-field solution and solid state NMR for characterisation of oil fractions responsible for wettability change in sandstones.

Asphaltene adsorption on solid surfaces is a standing problem in petroleum industry. It has an adverse effect on reservoir production and development by changing rock wettability, plugging pore throats, and affects oil transport through pipelines. Asphaltene chemistry constitutes important part of the ageing process as part of petrophysical studies and core analysis. The mechanisms and contribution of various oil components to adsorption processes is not fully understood. To investigate the kinetics of the ageing process and address the relative contribution of different oil components, we prepared three sets of sandstone core plugs aged in different oil mixtures over various time intervals. Cores were then re-saturated with decane to evaluate their wetting state using low-field NMR relaxometry by monitoring a change of surface relaxivity. Adsorbed deposits were then extracted from cores for solution-state NMR analysis. Their 1H and 1H-13C correlation spectra obtained using heteronuclear single quantum coherence (HSQC) technique were matched to spectra of four SARA (saturates, aromatics, resins and asphaltenes) components of oil mixtures to deduce components of deposits and inter-component interactions. We notice that wettability reversal of rock is inversely proportional to initial asphaltene concentration. Analysis of deposits reveals an increase in their aliphatic content over ageing time, which is accompanied by a change of the morphology of the pore space due to cluster aggregates forming a network. Results suggest that the ageing process in respect to the wetting state of rock samples consists of three distinctive stages: (i) an early-time period, when the fraction of most polar asphaltenes creates a discontinuous layer corresponding to mixed-wet state; (ii) an intermediate-time interval, at which the full grain coverage may be achieved (at favourable chemical environment) corresponding to strong oil-wetting; (iii) a late-time stage, where intense macro-aggregates accumulation occurs, changing the pore space integrity. It is likely asphaltene-aliphatic interactions leading to growth of sub-micron size macro-aggregates.

Discrete and Stereospecific Oligomers Prepared by Sequential and Alternating Single Unit Monomer Insertion.

Natural biopolymers, such as DNA and proteins, have uniform microstructures with defined molecular weight, precise monomer sequence, and stereoregularity along the polymer main chain that affords them unique biological functions. To reproduce such structurally perfect polymers and understand the mechanism of specific functions through chemical approaches, researchers have proposed using synthetic polymers as an alternative due to their broad chemical diversity and relatively simple manipulation. Herein, we report a new methodology to prepare sequence-controlled and stereospecific oligomers using alternating radical chain growth and sequential photoinduced RAFT single unit monomer insertion (photo-RAFT SUMI). Two families of cyclic monomers, the indenes and the N-substituted maleimides, can be alternatively inserted into RAFT agents, one unit at a time, allowing the monomer sequence to be controlled through sequential and alternating monomer addition. Importantly, the stereochemistry of cyclic monomer insertion into the RAFT agents is found to be trans-selective along the main chains due to steric hindrance from the repeating monomer units. All investigated cyclic monomers provide such trans-selectivity, but analogous acyclic monomers give a mixed cis- and trans-insertion.

LocMAP: A new localization method for the parametric processing of high resolution NMR data.

High resolution NMR spectroscopy offers a large number of data points that enable close peaks to be resolved. Data processing algorithms, however, have not yet been able to capitalize on this offering to achieve the highest permissible resolution. Although singular value decomposition (SVD) based methods such as matrix pencil (MPM) are theoretically able to achieve this, their onerous computational cost makes them impractical. In this work, we address this problem and propose a localized MPM method that we refer to as LocMaP, which is capable of delivering the promised high resolution while at the same time taking advantage of the computational efficiency of the FFT. We present the derivation of LocMaP and offer an efficient implementation of it. Evaluation using both Monte Carlo runs and a simulated FID establish the great potential of the proposed method.

Tuneable catechin functionalisation of carbohydrate polymers.

In this contribution, we present a strategy to functionalise three natural carbohydrate polymers (dextran - a neutral polymer, sodium alginate - an anionic polymer and chitosan - a cationic polymer) with catechin with excellent degrees of functionality. In a first step, the carbohydrate polymers were oxidised by sodium periodate to yield aldehyde functionalised carbohydrate polymers. The presence of aldehyde groups was exploited to attach catechin by an acid catalysed nucleophilic reaction. The degree of catechin functionalisation could be easily tuned by varying the acid concentration in the reaction mixture, achieving catechin functionalisation levels of up to 48% for dextran aldehyde catechin, 35% for chitosan-aldehyde-catechin and 22% for sodium alginate aldehyde catechin. 1H, 1H-13C HSQC and DOSY NMR were performed to elucidate the structural differences between the three aldehyde functionalised polysaccharides and how this affects their reactivity and conjugation behaviour. All three carbohydrate polymer-catechin conjugates showed superior free-radical scavenging activity compared with the non-functionalised polymers.

ß-Hydroxybutyrate Boosts Mitochondrial and Neuronal Metabolism but is not Preferred Over Glucose Under Activated Conditions.

The ketone body, ß-hydroxybutyrate (ßOHB), is metabolised by the brain alongside the mandatory brain fuel glucose. To examine the extent and circumstances by which ßOHB can supplement glucose metabolism, we studied guinea pig cortical brain slices using increasing concentrations of [U-13C]D-ßOHB in conjunction with [1-13C]D-glucose under conditions of normo- and hypoglycaemia, as well as under high potassium (40 mmol/L K+) depolarization in normo- and hypoglycaemic conditions. The contribution of ßOHB to synthesis of GABA was also probed by inhibiting the synthesis of glutamine, a GABA precursor, with methionine sulfoximine (MSO). [U-13C]D-ßOHB at lower concentrations (0.25 and 1.25 mmol/L) stimulated mitochondrial metabolism, producing greater total incorporation of label into glutamate and GABA but did not have a similar effect in the cytosolic compartment where labelling of glutamine was reduced at 1.25 mmol/L [U-13C]D-ßOHB. At higher concentrations (2.5 mmol/L) [U-13C]D-ßOHB inhibited metabolism of [1-13C]D-glucose, and reduced total label incorporation and total metabolite pools. When glucose levels were reduced, ßOHB was able to partially restore the loss of glutamate and GABA caused by hypoglycaemia, but was not able to supplement levels of lactate, glutamine or alanine or to prevent the increase in aspartate. Under depolarizing conditions glucose was the preferred substrate over ßOHB, even in hypoglycaemic conditions where comparatively less ßOHB was incorporated except into aspartate isotopomers. Inhibition of glutamine synthesis with MSO had no significant effect on incorporation of label from [U-13C]D-ßOHB into GABA C2,1 indicating that the majority of this GABA was synthesized in GABAergic neurons from [U-13C]D-ßOHB rather than from Gln C4,5 imported from astrocytes.

Fluorescent Glyco Single-Chain Nanoparticle-Decorated Nanodiamonds.

We introduce the light-induced collapse of single glycopolymer chains in water generating fluorescent glyco single-chain nanoparticles (SCNPs) and their subsequent functionalization onto nanodiamonds. The glycopolymer precursors are prepared by polymerizing an acetylated mannose-based methacrylate monomer followed by a deprotection and postpolymerization functionalization step, introducing profluorescent photoactive tetrazole groups and furan-protected maleimide moieties. Subsequent UV irradiation in highly diluted aqueous solution triggers intramolecular tetrazole-mediated cycloadditions, yielding glyco SCNPs featuring fluorescence as well as lectin binding properties. The obtained SCNPs are coated onto nanodiamonds by adsorption, and the obtained hybrid nanoparticles are in depth characterized in terms of size, functionality, and bioactivity. Different coating densities are achieved by altering the SCNP concentration. The prepared nanoparticles are nontoxic in mouse RAW 264.7 macrophages. Furthermore, the fluorescence of the SCNPs can be exploited to image the SCNP-coated nanodiamonds in macrophage cells via confocal fluorescence microscopy.

Campylobacter concisus utilizes blood but not short chain fatty acids despite showing associations with Firmicutes taxa.

Campylobacter concisus is a member of the oral microbiota that has been associated with the development of inflammatory bowel diseases. However, the role of the bacterium in disease aetiology remains poorly understood. Here, we examine optimal conditions for the growth of C. concisus, and the pathogenic potential of this bacterium in human gastrointestinal cells from the upper tract. Further, the presence of C. concisus in the lower tract of Crohn's disease (CD) patients undergoing therapy is observed, and the associations of C. concisus with the abundance of other microbial taxa and compounds they produce are evaluated. C. concisus strains had the ability to tolerate moderate levels of acidity, adhere to and invade esophageal and gastric cells; however, these properties did not correlate with their pathogenic potential in intestinal cells. The presence of the bacterium in the lower gut of CD patients was associated with an increased relative abundance of Faecalibacterium and Lachnospiraceae incertae sedis. Short chain fatty acids that can be produced by these microbial species did not appear to be responsible for this association. However, we identified genetic similarity between C. concisus and Firmicutes, specifically within aspartate and glutamate racemases. The potential pathogenesis of C. concisus in the upper gastrointestinal tract, and the responsiveness of the bacterium to therapy in a subset of CD patients warrant further investigation into whether this bacterium has a causal role in disease or its presence is incidental.

NMR spectroscopy to follow reaction progress in ionic liquids.

In order to understand reaction outcomes in ionic liquids, it is crucial to be able to follow the progress of these reactions. This review highlights the advantages of NMR spectroscopy over other analytical techniques in following reaction progress in ionic liquids, particularly addressing the practical aspects of the methodology and highlighting the range of processes that can be readily followed. Copyright © 2014 John Wiley & Sons, Ltd.

The dual-role of Pt(iv) complexes as active drug and crosslinker for micelles based on ß-cyclodextrin grafted polymer.

With a combination of RAFT and click chemistry an amphiphilic block copolymer with poly(ethylene glycol) methyl ether methacrylate (POEGMEMA) as hydrophilic block and poly(propargyl methacrylate) (PMA) as hydrophobic block has been successfully synthesized. To this, 6-azide-6-deoxy-ß-cyclodextrin (N3-ß-CD) was clicked creating a hosting environment for a hydrophobic small molecule platinum pro-drug. Oxoplatin, the oxidized version of cisplatin, has been modified on its axial ligand introducing cholic acid groups through esterification. This modified cisplatin forms a host-guest complex with ß-cyclodextrin that has been characterised via NMR spectroscopy. The host-guest interaction that the drug established with the ß-cyclodextrin grafted copolymer drove the self-assembly into nanoparticles of a diameter of 266 nm able to physical encapsulate the platinum-based drug. In the presence of ascorbic acid, 70% of the pro-drug is released over a period of 24 h. Cytotoxicity assays on ovarian cancer cells show that the polymer carrier improves the cytotoxicity of the platinum pro-drug. The IC50 value decreases from 37.7 µM with the pro-drug to 20.4 µM when the pro-drug is encapsulated into the polymer carrier. This is due to the fact that the uptake of the polymer carrier is up to 6 fold higher than the significantly smaller pro-drug by itself.

Putting corannulene in its place. Reactivity studies comparing corannulene with other aromatic hydrocarbons.

A series of aromatic hydrocarbons were investigated so as to compare the reactivity of corannulene with planar aromatic hydrocarbons. Corannulene was found to be more reactive than benzene, naphthalene and triphenylene to Friedel-Crafts acylation whilst electrophilic aromatic bromination was also used to confirm that triphenylene was less reactive than corannulene and that pyrene, perylene and acenaphthene were more so. The stabilisation of a neighbouring carbocation by the various aromatic systems was investigated through consideration of the rates of methanolysis of a series of benzylic alcohols. The reactivity series was found to parallel that observed for the electrophilic aromatic substitutions and both series are supported by computational studies. As such, a reactivity scale was devised that showed that corannulene was less reactive than would be expected for an aromatic planar species of similar pi electron count.

Silent information regulator 1 modulator resveratrol increases brain lactate production and inhibits mitochondrial metabolism, whereas SRT1720 increases oxidative metabolism.

Silent information regulators (SIRTs) have been shown to deacetylate a range of metabolic enzymes, including those in glycolysis and the Krebs cycle, and thus alter their activity. SIRTs require NAD(+) for their activity, linking cellular energy status to enzyme activity. To examine the impact of SIRT1 modulation on oxidative metabolism, this study tests the effect of ligands that are either SIRT-activating compounds (resveratrol and SRT1720) or SIRT inhibitors (EX527) on the metabolism of (13)C-enriched substrates by guinea pig brain cortical tissue slices with (13)C and (1)H nuclear magnetic resonance spectroscopy. Resveratrol increased lactate labeling but decreased incorporation of (13)C into Krebs cycle intermediates, consistent with effects on AMPK and inhibition of the F0/F1-ATPase. By testing with resveratrol that was directly applied to astrocytes with a Seahorse analyzer, increased glycolytic shift and increased mitochondrial proton leak resulting from interactions of resveratrol with the mitochondrial electron transport chain were revealed. SRT1720, by contrast, stimulated incorporation of (13)C into Krebs cycle intermediates and reduced incorporation into lactate, although the inhibitor EX527 paradoxically also increased Krebs cycle (13)C incorporation. In summary, the various SIRT1 modulators show distinct acute effects on oxidative metabolism. The strong effects of resveratrol on the mitochondrial respiratory chain and on glycolysis suggest that caution should be used in attempts to increase bioavailability of this compound in the CNS.

Competitive formation of DNA linkage isomers by a trinuclear platinum complex and the influence of pre-association.

2D [(1)H, (15)N] HSQC NMR spectroscopy has been used to monitor the reaction of fully (15)N-labelled [{trans-PtCl(NH3)2}2(µ-trans-Pt(NH3)2{NH2(CH2)6NH2}2)](4+) (BBR3464 ((15)N-1)) with the 14-mer duplex (5'-{d(ATACATG(7)G(8)TACATA)}-3'·5'-{d(TATG(18)TACCATG(25)TAT)}-3' or I) at pH 5.4 and 298 K, to examine the possible formation of 1,4 and 1,5-GG adducts in both 5'-5' and 3'-3' directions. In a previous study, the binding of the dinuclear 1,1/t,t to I showed specific formation of the 5'-5' 1,4 G(8)G(18) cross-link, whereas in this case a mixture of adducts were formed. Initial (1)H NMR spectra suggested the presence of two pre-associated states aligned in both directions along the DNA. The pre-association was studied in the absence of covalent binding, by use of the "non-covalent" analog [{trans-Pt(NH3)3}2(µ-trans-Pt(NH3)2{NH2(CH2)6NH2}2)](6+) (AH44, 0). Chemical shift changes of DNA protons combined with NOE connectivities between CH2 and NH3 protons of 0 and the adenine H2 protons on I show that two different molecules of 0 are bound in the minor groove. Molecular dynamic simulations were performed to study the interaction of 0 at the two pre-association sites using charges derived from density functional theory (DFT) calculations. Structures where the central platinum is located in the minor groove and the aliphatic linkers extend into the major groove, in opposite directions, often represent the lowest energy structures of the snapshots selected. In the reaction of (15)N-1 and I, following the pre-association step, aquation occurs to give the mono aqua monochloro species 2, with a rate constant of 3.43 ± 0.03 × 10(-5) s(-1). There was evidence for two monofunctional adducts (3, 4) bound to the 3' (G8) and 5' (G7) residues and the asymmetry of the (1)H,(15)N peak for 3 suggested two conformers of the 3' adduct, aligned in different directions along the DNA. The rate constant for combined monofunctional adduct formation (0.6 ± 0.1 M(-1)) is ca. 2-fold lower for 1 compared to 1,1/t,t, whereas the rate constant for conversion of the combined monofunctional species to combined bifunctional adducts (5) (8.0 ± 0.2 × 10(-5) s(-1)) is two-fold higher.