Granulocyte transfusion during cord blood transplant for relapsed, refractory AML is associated with massive CD8 T-cell expansion, significant cytokine release syndrome and induction of disease remission.

In high-risk myeloid malignancy, relapse is reduced using cord blood transplant (CBT) but remains the principal cause of treatment failure. We previously described T-cell expansion in CBT recipients receiving granulocyte transfusions. We now report the safety and tolerability of such transfusions, T-cell expansion data, immunophenotype, cytokine profiles and clinical response in children with post-transplant relapsed acute leukaemia who received T-replete, HLA-mismatched CBT and pooled granulocytes within a phase I/II trial (ClinicalTrials.

Synthesis and antimicrobial activity of an SO-releasing siderophore conjugate.

A novel Trojan Horse conjugate consisting of an SO-releasing 2,4-dinitrobenzenesulfonamide group attached to the monocatecholate siderophore aminochelin was synthesized to examine whether a bidentate catecholate siderophore unit could help potentiate the antimicrobial activity of SO-releasing prodrugs. The conjugate obtained displays rapid SO release on reaction with glutathione, and proved more active against Staphylococcus aureus than a comparable SO-releasing prodrug lacking the siderophore unit, although activity required micromolar concentrations. The conjugate was inactive against wild-type Escherichia coli, but activity was observed against an entA mutant strain that is unable to produce its major siderophores.

A Salmochelin S4-Inspired Ciprofloxacin Trojan Horse Conjugate.

A novel ciprofloxacin-siderophore Trojan Horse antimicrobial was prepared by incorporating key design features of salmochelin, a stealth siderophore that evades mammalian siderocalin capture its glycosylated catechol units. Assessment of the antimicrobial activity of the conjugate revealed that attachment of the salmochelin mimic resulted in decreased potency, compared to ciprofloxacin, against two strains, K12 and Nissle 1917, in both iron replete and deplete conditions. This observation could be attributed to a combination of reduced DNA gyrase inhibition, as confirmed by DNA gyrase assays, and reduced bacterial uptake.

Investigation of Parameters that Affect Resin Swelling in Green Solvents.

The influence of various physical and chemical factors on the swelling of polystyrene and PEG based resins in greener organic solvents has been systematically investigated. In general, chemical factors: the nature of the functionality/linker and the degree of loading were found to have a far larger influence on the swelling of the resins than physical parameters such as bead size. The results are interpreted in terms of Hansen solubility parameters for the solvents and there is evidence that some solvents interact with the polymeric core of a resin whilst others interact with the functionality.

Mimicking salmochelin S1 and the interactions of its Fe(III) complex with periplasmic iron siderophore binding proteins CeuE and VctP.

A mimic of the tetradentate stealth siderophore salmochelin S1, was synthesised, characterised and shown to form Fe(III) complexes with ligand-to-metal ratios of 1:1 and 3:2. Circular dichroism spectroscopy confirmed that the periplasmic binding proteins CeuE and VctP of Campylobacter jejuni and Vibrio cholerae, respectively, bind the Fe(III) complex of the salmochelin mimic by preferentially selecting Λ-configured Fe(III) complexes. Intrinsic fluorescence quenching studies revealed that VctP binds Fe(III) complexes of the mimic and structurally-related catecholate ligands, such as enterobactin, bis(2, 3-dihydroxybenzoyl-l-serine) and bis(2, 3-dihydroxybenzoyl)-1, 5-pentanediamine with higher affinity than does CeuE.

Chemical and Biological Aspects of Nutritional Immunity-Perspectives for New Anti-Infectives that Target Iron Uptake Systems.

Upon bacterial infection, one of the defense mechanisms of the host is the withdrawal of essential metal ions, in particular iron, which leads to "nutritional immunity". However, bacteria have evolved strategies to overcome iron starvation, for example, by stealing iron from the host or other bacteria through specific iron chelators with high binding affinity. Fortunately, these complex interactions between the host and pathogen that lead to metal homeostasis provide several opportunities for interception and, thus, allow the development of novel antibacterial compounds.

Fabrication and Application of Isotopically Labelled Gold Arrays for Multiplexed Peptide Analysis.

A new array-based technology for the simultaneous capture, chemical labelling and mass spectrometry analysis of peptides is presented. Isotopically labelled self-assembled monolayer (SAM) gold arrays are constructed and used simultaneously to capture and label a range of peptides. The array-immobilised, labelled peptides were released by MALDI ablation, analysed by MALDI mass spectrometry and readily identified as labelled peptides from their characteristic isotope pattern.

Probing linker design in citric acid-ciprofloxacin conjugates.

A series of structurally related citric acid-ciprofloxacin conjugates was synthesised to investigate the influence of the linker between citric acid and ciprofloxacin on antibacterial activities. Minimum inhibitory concentrations (MICs) were determined against a panel of reference strains and clinical isolates of bacteria associated with infection in humans and correlated with the DNA gyrase inhibitory activity. The observed trend was rationalised by computational modelling.

Probing bacterial uptake of glycosylated ciprofloxacin conjugates.

Mono- and disaccharide-functionalised conjugates of the fluoroquinolone antibiotic ciprofloxacin have been synthesised and used as chemical probes of the bacterial uptake of glycosylated ciprofloxacin. Their antimicrobial activities against a panel of clinically relevant bacteria were determined: the ability of these conjugates to inhibit their target DNA gyrase and to be transported into the bacteria was assessed by using in vivo and in vitro assays. The data suggest a lack of active uptake through sugar transporters and that although the addition of monosaccharides is compatible with the inhibition of DNA gyrase, the addition of a disaccharide results in a significant decrease in antimicrobial activity.

Staphyloferrin A as siderophore-component in fluoroquinolone-based Trojan horse antibiotics.

A series of fluoroquinolone conjugates was synthesised by linking the carboxylic acid functionality of the carboxylate-type siderophore staphyloferrin A and its derivatives to the piperazinyl nitrogen of ciprofloxacin and norfloxacin via amide bond formation. Four siderophore-drug conjugates were screened against a panel of bacteria associated with infection in humans. Whilst no activity was found against ciprofloxacin- or norfloxacin-resistant bacteria, one of the conjugates retained antibacterial activity against fluoroquinolone-susceptible strains although the structure of its lysine-based siderophore component differs from that of the natural siderophore staphyloferrin A.

Evaluation of a solid-supported tagging strategy for mass spectrometric analysis of peptides.

We have explored two divinylbenzene cross-linked polystyrene supports for use in a solid-supported N-terminal peptide tagging strategy. Resin-bound tags designed to be cleaved in a single step at the N-terminus of peptides have been devised and explored as peptide N-terminal tagging reagents (constructs) for subsequent mass spectrometric analysis. While the brominated tagging approach shows promise, the use of these specific solid supports has drawbacks, in terms of tagging reaction scale, for real applications in proteomics.

Dietary intake estimate for perfluorooctanesulphonic acid (PFOS) and other perfluorocompounds (PFCs) in UK retail foods following determination using standard addition LC-MS/MS.

The analysis of 252 food samples (UK-produced and imported) purchased from a variety of retail outlets in the UK was undertaken for the presence of perfluorooctanesulphonic acid (PFOS), perfluorooctanoic acid (PFOA) and nine other perfluorocompounds (PFCs). A limit of quantification (LOQ) of 1 microg/kg was achieved for all target analytes, in all samples. Standard addition was used for quantification of PFC levels.

Mass spectral studies towards more reliable measurement of perfluorooctanesulfonic acid and other perfluorinated chemicals (PFCs) in food matrices using liquid chromatography/tandem mass spectrometry.

Liquid chromatography/mass spectrometry (LC/MS) experiments are described, leading to a reliable method for the measurement of perfluorooctanesulfonic acid (PFOS) and other perfluorinated chemicals (PFCs) in foods. Separations were performed on new fluorinated stationary phases, RP Octyl (-C(8)F(17)) or propyl-perfluorobenzene (-C(3)H(6)-C(6)F(5)), to ensure resolution of PFOS and interfering taurohydroxycholate isomers. Aqueous ammonium formate (5 mM) and methanol were used as the mobile phases.

Synthesis of citrate-ciprofloxacin conjugates.

Two regioisomeric citrate-functionalized ciprofloxacin conjugates have been synthesized and their antimicrobial activities against a panel of clinically-relevant bacteria have been determined. Cellular uptake mechanisms were investigated using wild-type and ompF deletion strains of Escherichia coli K-12.

Characterisation and proposed origin of mass spectrometric ions observed 30 Th above the ionised molecules of per-O-methylated carbohydrates.

Derivatisation of carbohydrates by permethylation significantly improves the mass spectrometric intensity of carbohydrate-derived ions and allows more readily interpretable fragmentation; in addition, samples are conveniently separated from salts, and larger oligosaccharides are more readily ionised. It has previously been recognised that, in the mass spectra of permethylated carbohydrates, a series of ions indicating species 30 Da larger than the fully methylated carbohydrate molecules are also observed. These species have not been characterised in the literature despite their apparently ubiquitous occurrence in the mass spectra of permethylated carbohydrates.

Recent advances in analytical construct resins.

The area of solid-phase synthesis has witnessed exponential growth in the last fifteen years, but difficulties associated with the monitoring and analysis of resin-bound reactions and products have been apparent due to a limited number of analytical methods available. With the substrate tethered to an insoluble support traditional chromatographic monitoring is only possible after cleavage. In order to address this 'analytical bottleneck' Geysen, in 1996 elaborated Merrifield's initial dual linker strategy by incorporating an encoding system between two in-line linkers.

Characterization of a proteinase inhibitor from Brachypodium distachyon suggests the conservation of defence signalling pathways between dicotyledonous plants and grasses.

SUMMARY Proteinase inhibitors (PIs) are established markers for wound- and especially jasmonate-mediated signalling in dicot species such as tomato and potato. Differential screening of a cDNA library constructed from RNA isolated from wounded leaves of the grass Brachypodium distachyon led to the identification of a proteinase inhibitor gene (Bdpin1). Bdpin1 exhibited the highest homology to the subtilisin/chymotrypsin-inhibiting subgroup of the pin1 class of plant PIs.

Magnaporthe grisea interactions with the model grass Brachypodium distachyon closely resemble those with rice (Oryza sativa).

SUMMARY Germplasm of Brachypodium distachyon was inoculated with Magnaporthe grisea using either rice- (Guy11) or grass-adapted (FAG1.1.1, PA19w-06, PA31v-01) host-limited forms of the fungus, and interactions with varying degrees of susceptibility and resistance were identified.

Brachypodium distachyon. A new model system for functional genomics in grasses.

A new model for grass functional genomics is described based on Brachypodium distachyon, which in the evolution of the Pooideae diverged just prior to the clade of "core pooid" genera that contain the majority of important temperate cereals and forage grasses. Diploid ecotypes of B. distachyon (2n = 10) have five easily distinguishable chromosomes that display high levels of chiasma formation at meiosis.

Surface stability of granular systems under horizontal and vertical vibration: the applicability of a coefficient of friction.

We investigate the conditions under which the surface of a granular pile becomes unstable to vibrations. Three stability boundaries are identified, which depend upon the relative phase of the driving forces and the angle of the prepared slope. The experimental findings can be interpreted within the context of a Coulomb friction model and used to define an effective coefficient of friction.

The glycosidic precursor of (Z)-5-ethylidene-2(5H)-furanone in Halocarpus biformis juvenile foliage.

A new glycosidic lactone, (5R,6R)-5-(1-hydroxyethyl)-2(5H)-furanone beta-D-glucopyranoside, has been identified as the principal precursor of (Z)-5-ethylidene-2(5H)-furanone in juvenile foliage of the New Zealand tree Halocarpus biformis. Three related lactone glycosides were isolated in smaller amounts, together with the known phenolic glycosides pyroside, arbutin and picein. The principal lactone glycoside underwent facile elimination of glucose, in neutral or basic conditions, to yield (Z)-5-ethylidene-2(5H)-furanone and its E-isomer.

Identification and estimation of cell types in mixed primary cell cultures of early and term human placenta.

Placental villi of early and term human placentae were dissociated with trypsin and mixed cell cultures were established. The different cell types were identified and estimated over a 14-day culture period using antibodies to keratin and vimentin filaments and their capacity to phagocytose yeast. The three main cell types were found to be epithelial-, macrophage- and fibroblast-like cells.

The effect of type and dose of oestrogen on the blood pressure of post-menopausal women.

Evidence from two groups of post-menopausal women who were randomly allocated to be treated with either conjugated equine oestrogens or piperazine oestrone sulphate demonstrates that the two oestrogens produce markedly different effects on blood pressure. The conjugated equine oestrogens appear to produce no significant change in either systolic or diastolic blood pressure whereas piperazine oestrone sulphate produced a significant fall in both systolic and diastolic blood pressure. This finding is discussed in relationship to the known causes for a change in cardiovascular response to oestrogen and several hypotheses are put forward.