Exploring Design Concepts for Siderophore-Fluoroquinolone Trojan Horse Antimicrobials

Urgent action is required to combat the ongoing threat of antimicrobial resistance. Trojan horse conjugates, where antimicrobials are linked to a nutrient carrier, can evade permeability-related resistance through active transport of drugs into bacterial cells. The work presented herein explores modification of the linker moiety between a citrate siderophore and ciprofloxacin, as well as the addition of glycosyl groups to catecholate siderophore moieties to mimic the salmochelins; stealth siderophores which can evade the mammalian defence protein siderocalin. Two linkers were investigated, one containing a carbamate and the other a disulfide bond neighbouring a carbamate group. These were chosen to give intracellular release of the antimicrobial, either by the action of carboxylesterases or through reduction of the disulfide bond by thiolate anions in the cytoplasm, respectively. A carbamate-linked conjugate was synthesised and screened against wild type E. coli, and demonstrated lower antimicrobial activity than that for the parent drug. Screening against a bacterial strain lacking the outer membrane ferric citrate receptor FecA demonstrated that FecA is not essential for uptake of the synthesised conjugate. The synthesis of a disulfide linked conjugate was unsuccessful, due to release of free ciprofloxacin during the final deprotection step. A salmochelin-inspired conjugate was synthesised. Whilst it demonstrated reduced antimicrobial activity against wild type E. coli in iron rich media, compared to the parent drug, higher antimicrobial activity was observed in iron-limited media, suggesting active uptake. A DNA gyrase assay showed that the inhibitory activity of this conjugate was lower than the free drug, suggesting that the antimicrobial activity observed under iron deficiency may be due extracellular iron sequestration by the siderophore

Clusters of Galaxies: New Results from the CLEF Hydrodynamics Simulation

Preliminary results are presented from the CLEF hydrodynamics simulation, a large (N=2(428)^3 particles within a 200 Mpc/h comoving box) simulation of the LCDM cosmology that includes both radiative cooling and a simple model for galactic feedback. Specifically, we focus on the X-ray properties of the simulated clusters at z=0 and demonstrate a reasonable level of agreement between simulated and observed cluster scaling relations.Comment: 7 pages, 4 figures, accepted for publication in Advances in Space Research (proceedings of the COSPAR 2004 Assembly, Paris

Thermal stability of dialkylimidazolium tetrafluoroborate and hexafluorophosphate ionic liquids: ex situ bulk heating to complement in situ mass spectrometry

Thermal decomposition (TD) products of the ionic liquids (ILs) [CnC1Im][BF4] and [CnC1Im][PF6] ([CnC1Im]+ = 1-alkyl-3-methylimidazolium, [BF4]- = tetrafluoroborate, and [PF6]- = hexafluorophosphate) were prepared, ex situ, by bulk heating experiments in a bespoke setup. The respective products, CnC1(C3N2H2)BF3 and CnC1(C3N2H2)PF5 (1-alkyl-3-methylimidazolium-2-trifluoroborate and 1-alkyl-3-methylimidazolium-2-pentafluorophosphate), were then vaporized and analyzed by direct insertion mass spectrometry (DIMS) in order to identify their characteristic MS signals. During IL DIMS experiments we were subsequently able, in situ, to identify and monitor signals due to both IL vaporization and IL thermal decomposition. These decomposition products have not been observed in situ during previous analytical vaporization studies of similar ILs. The ex situ preparation of TD products is therefore perfectly complimentary to in situ thermal stability measurements. Experimental parameters such as sample surface area to volume ratios and heating rates are consequently very important for ILs that show competitive vaporization and thermal decomposition. We have explained these experimental factors in terms of Langmuir evaporation and Knudsen effusion-like conditions, allowing us to draw together observations from previous studies to make sense of the literature on IL thermal stability. Hence, the design of experimental setups are crucial and previously overlooked experimental factors

Alternative job search strategies in remote rural and peri-urban labour markets: the role of social networks

This paper examines the importance of informal methods (especially social networking) to the job search strategies used by unemployed people. It compares three areas: a small rural town; a larger, more sparsely populated, remote rural area; and a centrally-located, peri-urban labour market. The analysis is based first on survey research undertaken with 490 job seekers across the study areas. Emerging issues were then followed up during a series of twelve focus groups. The survey research showed that job seekers in the rural study areas were significantly more likely to use social networks to look for work. However, those who had experienced repeated or long-term periods out of work, the unskilled and young people were significantly less likely to use such networks. Focus groups confirmed the perceived importance of social networking to the job search process in rural areas, in contrast to the more marginal role such methods appear to play in peri-urban settings. For many rural job seekers, formal job search activities conducted through Jobcentres were seen as largely symbolic, lacking the practical value of social networking. These results suggest that service providers seeking to assist unemployed people in rural areas need to address the problems faced by many disadvantaged job seekers who are currently caught between their lack of social network relations and the absence of local public employment service facilities in more remote communities

3,3'-Diindolylmethane (DIM) and its ring-substituted halogenated analogs (ring-DIMs) induce differential mechanisms of survival and death in androgen-dependent and -independent prostate cancer cells.

International audienceWe recently reported that novel ring-substituted analogs of 3,3'-diindolylmethane (ring-DIMs) induce apoptosis and necrosis in androgen-dependent and -independent prostate cancer cells. In this paper, we have focused on the mechanism(s) associated with ring-DIM-mediated cell death, and on identifying the specific intracellular target(s) of these compounds. The 4,4'- and 7,7'-dichloroDIMs and 4,4'- and 7,7'-dibromoDIMs induced the death of LNCaP, C42B and DU145 prostate cancer cells, but not that of immortalized normal human prostate epithelial (RWPE-1) cells. Ring-DIMs caused the early loss of mitochondrial membrane potential (MMP) and decreased mitochondrial ATP generation in prostate cancer cells. Cyclosporin A, an inhibitor of the mitochondrial permeability transition pore, inhibited ring-DIM-mediated cell death, and salubrinal, an inhibitor of ER stress, inhibited cell death mediated only by 4,4'-dihaloDIMs. We found that although salubrinal did not inhibit the onset of ER stress, it prevented 4,4'-dibromoDIM mediated loss of MMP. Salubrinal potentiated cell death in response to 7,7'-dihaloDIMs and DIM, and this effect concurred with increased loss of MMP. Using in silico 3-D docking affinity analysis, we identified Ca2+/calmodulin-dependent kinase II (CaMKII) as a potential direct target for the most toxic ring-DIM, 4,4'-dibromoDIM. An inhibitor of CaMKII, KN93, but not its inactive analog KN92, abrogated cell death mediated by 4,4'-dibromoDIM. The ring-DIMs induced ER stress and autophagy, but these processes were not necessary for ring-DIM-mediated cell death. Inhibition of autophagy with bafilomycin A1, 3-methyladenine or by LC3B gene silencing sensitized LNCaP and C42B, but not ATG5-deficient DU145 cells to ring-DIM- and DIM-mediated cell death. We propose that autophagy induced by the ring-DIMs and DIM has a cytoprotective function in prostate cancer cells

Glutamatergic dysfunction leads to a hyper-dopaminergic phenotype through deficits in short-term habituation: a mechanism for aberrant salience

Psychosis in disorders like schizophrenia is commonly associated with aberrant salience and elevated striatal dopamine. However, the underlying cause(s) of this hyper-dopaminergic state remain elusive. Various lines of evidence point to glutamatergic dysfunction and impairments in synaptic plasticity in the etiology of schizophrenia, including deficits associated with the GluA1 AMPAR subunit. GluA1 knockout (Gria1−/−) mice provide a model of impaired synaptic plasticity in schizophrenia and exhibit a selective deficit in a form of short-term memory which underlies short-term habituation. As such, these mice are unable to reduce attention to recently presented stimuli. In this study we used fast-scan cyclic voltammetry to measure phasic dopamine responses in the nucleus accumbens of Gria1−/− mice to determine whether this behavioral phenotype might be a key driver of a hyper-dopaminergic state. There was no effect of GluA1 deletion on electrically-evoked dopamine responses in anaesthetized mice, demonstrating normal endogenous release properties of dopamine neurons in Gria1−/− mice. Furthermore, dopamine signals were initially similar in Gria1−/− mice compared to controls in response to both sucrose rewards and neutral light stimuli. They were also equally sensitive to changes in the magnitude of delivered rewards. In contrast, however, these stimulus-evoked dopamine signals failed to habituate with repeated presentations in Gria1−/− mice, resulting in a task-relevant, hyper-dopaminergic phenotype. Thus, here we show that GluA1 dysfunction, resulting in impaired short-term habituation, is a key driver of enhanced striatal dopamine responses, which may be an important contributor to aberrant salience and psychosis in psychiatric disorders like schizophrenia

Cortical cells are altered by factors including bone morphogenetic protein released from a placental barrier model under altered oxygenation

Episodes of hypoxia and hypoxia/reoxygenation during foetal development have been associated with increased risk of neurodevelopmental conditions presenting in later life. The mechanism for this is not understood; however, several authors have suggested that the placenta plays an important role. Previously we found both placentas from a maternal hypoxia model and pre-eclamptic placentas from patients release factors lead to a loss of dendrite complexity in rodent neurons. Here to further explore the nature and origin of these secretions we exposed a simple in vitro model of the placental barrier, consisting of a barrier of human cytotrophoblasts, to hypoxia or hypoxia/reoxygenation. We then exposed cortical cultures from embryonic rat brains to the conditioned media (CM) from below these exposed barriers and examined changes in cell morphology, number, and receptor presentation. The barriers released factors that reduced dendrite and astrocyte process lengths, decreased GABAB1 staining, and increased astrocyte number. The changes in astrocytes required the presence of neurons and were prevented by inhibition of the SMAD pathway and by neutralising Bone Morphogenetic Proteins (BMPs) 2/4. Barriers exposed to hypoxia/reoxygenation also released factors that reduced dendrite lengths but increased GABAB1 staining. Both oxygen changes caused barriers to release factors that decreased GluN1, GABAAα1 staining and increased GluN3a staining. We find that hypoxia in particular will elicit the release of factors that increase astrocyte number and decrease process length as well as causing changes in the intensity of glutamate and GABA receptor staining. There is some evidence that BMPs are released and contribute to these changes

The dark haloes of early-type galaxies in low-density environments: XMM-Newton and Chandra observations of NGC 57, NGC 7796 and IC 1531

We present analysis of Chandra and XMM-Newton observations of three early-type galaxies, NGC 57, NGC 7796 and IC 1531. All three are found in very low density environments, and appear to have no neighbours of comparable size. NGC 57 has a halo of kT~0.9 keV, solar metallicity gas, while NGC 7796 and IC 1531 both have ~0.55 keV, 0.5-0.6 Zsol haloes. IC 1531 has a relatively compact halo, and we consider it likely that gas has been removed from the system by the effects of AGN heating. For NGC 57 and NGC 7796 we estimate mass, entropy and cooling time profiles and find that NGC 57 has a fairly massive dark halo with a mass-to-light ratio of 44.7 (4.0,-8.5) Msol/Lsol (1 sigma uncertainties) at 4.75 Re. This is very similar to the mass-to-light ratio found for NGC 4555 and confirms that isolated ellipticals can possess sizable dark matter haloes. We find a significantly lower mass-to-light ratio for NGC 7796, 10.6 (+2.5,-2.3) Msol/Lsol at 5 Re, and discuss the possibility that NGC 7796 hosts a galactic wind, causing us to underestimate its mass.Comment: 14 pages, 9 figures, accepted for publication in MNRA