Macrophage TNF-α mediates parathion-induced airway hyperreactivity in guinea pigs.

Organophosphorus pesticides (OPs) are implicated in human asthma. We previously demonstrated that, at concentrations that do not inhibit acetylcholinesterase activity, the OP parathion causes airway hyperreactivity in guinea pigs as a result of functional loss of inhibitory M2 muscarinic receptors on parasympathetic nerves. Because macrophages are associated with asthma, we investigated whether macrophages mediate parathion-induced M2 receptor dysfunction and airway hyperreactivity. Airway physiology was measured in guinea pigs 24 h after a subcutaneous injection of parathion. Pretreatment with liposome-encapsulated clodronate induced alveolar macrophage apoptosis and prevented parathion-induced airway hyperreactivity in response to electrical stimulation of the vagus nerves. As determined by qPCR, TNF-α and IL-1β mRNA levels were increased in alveolar macrophages isolated from parathion-treated guinea pigs. Parathion treatment of alveolar macrophages ex vivo did not significantly increase IL-1β and TNF-α mRNA but did significantly increase TNF-α protein release. Consistent with these data, pretreatment with the TNF-α inhibitor etanercept but not the IL-1β receptor inhibitor anakinra prevented parathion-induced airway hyperreactivity and protected M2 receptor function. These data suggest a novel mechanism of OP-induced airway hyperreactivity in which low-level parathion activates macrophages to release TNF-α-causing M2 receptor dysfunction and airway hyperreactivity. These observations have important implications regarding therapeutic approaches for treating respiratory disease associated with OP exposures

The Local Group as a test of cosmological models

The dynamics of the Local Group and its environment provide a unique challenge to cosmological models. The velocity field within 5h-1 Mpc of the Local Group (LG) is extremely ``cold''. The deviation from a pure Hubble flow, characterized by the observed radial peculiar velocity dispersion, is measured to be about 60km/s. We compare the local velocity field with similarly defined regions extracted from N-body simulations of Universes dominated by cold dark matter (CDM). This test is able to strongly discriminate between models that have different mean mass densities. We find that neither the Omega=1 (SCDM) nor Omega=0.3 (OCDM) cold dark matter models can produce a single candidate Local Group that is embedded in a region with such small peculiar velocities. For these models, we measure velocity dispersions between 500-700km/s and 150-300km/s respectively, more than twice the observed value. Although both CDM models fail to produce environments similar to those of our Local Group on a scale of a few Mpc, they can give rise to many binary systems that have similar orbital properties as the Milky Way--Andromeda system. The local, gravitationally induced bias of halos in the CDM ``Local Group'' environment, if defined within a sphere of 10 Mpc around each Local Group is about 1.5, independent of Omega. No biasing scheme could reconcile the measured velocity dispersions around Local Groups with the observed one. Identification of binary systems using a halo finder (named Skid (http://www-hpcc.astro.washington.edu/tools/DENMAX for a public version)) based on local density maxima instead of a simple linking algorithm, gives a much more complete sample. We show that a standard ``friend of friends'' algorithm would miss 40% of the LG candidates present in the simulations.Comment: Latex file (19 pages) + 13 figures. Submitted to New Astronomy. Two MPEG movies were not included. Also available (this time with the movies) at http://www-hpcc.astro.washington.edu/faculty/fabio/index.htm

The metallicity gradient as a tracer of history and structure : the Magellanic Clouds and M33 galaxies

Original article can be found at: http://www.aanda.org/ Copyright The European Southern Observatory (ESO) DOI: 10.1051/0004-6361/200912138Context. The stellar metallicity and its gradient place constraints on the formation and evolution of galaxies. Aims. This is a study of the metallicity gradient of the LMC, SMC and M33 galaxies derived from their asymptotic giant branch (AGB) stars. Methods. The [Fe/H] abundance was derived from the ratio between C- and M-type AGB stars and its variation analysed as a function of galactocentric distance. Galaxy structure parameters were adopted from the literature. Results. The metallicity of the LMC decreases linearly as −0.047±0.003 dex kpc−1 out to ∼8 kpc from the centre. In the SMC, [Fe/H] has a constant value of ∼−1.25 ± 0.01 dex up to ∼12 kpc. The gradient of the M33 disc, until ∼9 kpc, is −0.078 ± 0.003 dex kpc−1 while the outer disc/halo, out to ∼25 kpc, has [Fe/H] ∼ −1.7 dex. Conclusions. The metallicity of the LMC, as traced by different populations, bears the signature of two major star forming episodes: the first one constituting a thick disc/halo population and the second one a thin disc and bar due to a close encounter with the Milky Way and SMC. The [Fe/H] of the recent episode supports an LMC origin for the Stream. The metallicity of the SMC supports star formation, ∼3 Gyr ago, as triggered by LMC interaction and sustained by the bar in the outer region of the galaxy. The SMC [Fe/H] agrees with the present-day abundance in the Bridge and shows no significant gradient. The metallicity of M33 supports an “insideout” disc formation via accretion of metal poor gas from the interstellar medium.Peer reviewe

Simultaneous quantification of 12 different nucleotides and nucleosides released from renal epithelium and in human urine samples using ion-pair reversed-phase HPLC

Nucleotides and nucleosides are not only involved in cellular metabolism but also act extracellularly via P1 and P2 receptors, to elicit a wide variety of physiological and pathophysiological responses through paracrine and autocrine signalling pathways. For the first time, we have used an ion-pair reversed-phase high-performance liquid chromatography ultraviolet (UV)-coupled method to rapidly and simultaneously quantify 12 different nucleotides and nucleosides (adenosine triphosphate, adenosine diphosphate, adenosine monophosphate, adenosine, uridine triphosphate, uridine diphosphate, uridine monophosphate, uridine, guanosine triphosphate, guanosine diphosphate, guanosine monophosphate, guanosine): (1) released from a mouse renal cell line (M1 cortical collecting duct) and (2) in human biological samples (i.e., urine). To facilitate analysis of urine samples, a solid-phase extraction step was incorporated (overall recovery rate ? 98 %). All samples were analyzed following injection (100 ?l) into a Synergi Polar-RP 80 Å (250 × 4.6 mm) reversed-phase column with a particle size of 10 ?m, protected with a guard column. A gradient elution profile was run with a mobile phase (phosphate buffer plus ion-pairing agent tetrabutylammonium hydrogen sulfate; pH 6) in 2-30 % acetonitrile (v/v) for 35 min (including equilibration time) at 1 ml min(-1) flow rate. Eluted compounds were detected by UV absorbance at 254 nm and quantified using standard curves for nucleotide and nucleoside mixtures of known concentration. Following validation (specificity, linearity, limits of detection and quantitation, system precision, accuracy, and intermediate precision parameters), this protocol was successfully and reproducibly used to quantify picomolar to nanomolar concentrations of nucleosides and nucleotides in isotonic and hypotonic cell buffers that transiently bathed M1 cells, and urine samples from normal subjects and overactive bladder patients

The porin and the permeating antibiotic: A selective diffusion barrier in gram-negative bacteria

Gram-negative bacteria are responsible for a large proportion of antibiotic resistant bacterial diseases. These bacteria have a complex cell envelope that comprises an outer membrane and an inner membrane that delimit the periplasm. The outer membrane contains various protein channels, called porins, which are involved in the influx of various compounds, including several classes of antibiotics. Bacterial adaptation to reduce influx through porins is an increasing problem worldwide that contributes, together with efflux systems, to the emergence and dissemination of antibiotic resistance. An exciting challenge is to decipher the genetic and molecular basis of membrane impermeability as a bacterial resistance mechanism. This Review outlines the bacterial response towards antibiotic stress on altered membrane permeability and discusses recent advances in molecular approaches that are improving our knowledge of the physico-chemical parameters that govern the translocation of antibiotics through porin channel

Memory for medication side effects in younger and older adults: the role of subjective and objective importance

Older adults often experience memory impairments, but can sometimes use selective processing and schematic support to remember important information. The current experiments investigate to what degree younger and healthy older adults remember medication side effects that were subjectively or objectively important to remember. Participants studied a list of common side effects, and rated how negative these effects were if they were to experience them, and were then given a free recall test. In Experiment 1, the severity of the side effects ranged from mild (e.g., itching) to severe (e.g., stroke), and in Experiment 2, certain side effects were indicated as critical to remember (i.e., “contact your doctor if you experience this”). There were no age differences in terms of free recall of the side effects, and older adults remembered more severe side effects relative to mild effects. However, older adults were less likely to recognize critical side effects on a later recognition test, relative to younger adults. The findings suggest that older adults can selectively remember medication side effects, but have difficulty identifying familiar but potentially critical side effects, and this has implications for monitoring medication use in older age

Divided attention selectively impairs memory for self-relevant information

Information that is relevant to oneself tends to be remembered more than information that relates to other people, but the role of attention in eliciting this "self-reference effect" is unclear. In the present study, we assessed the importance of attention in self-referential encoding using an ownership paradigm, which required participants to encode items under conditions of imagined ownership by themselves or by another person. Previous work has established that this paradigm elicits a robust self-reference effect, with more "self-owned" items being remembered than "other-owned" items. Access to attentional resources was manipulated using divided-attention tasks at encoding. A significant self-reference effect emerged under full-attention conditions and was related to an increase in episodic recollection for self-owned items, but dividing attention eliminated this memory advantage. These findings are discussed in relation to the nature of self-referential cognition and the importance of attentional resources at encoding in the manifestation of the self-reference effect in memory

Planetary nebulae: abundances and abundance gradients

In this work, a review is given of some recent results and problems involved in the determination of chemical abundances of galactic planetary nebulae, particularly regarding disk and bulge objects

Rapid assembly of customized TALENs into multiple

Transcriptional activator-like effector nucleases (TALENs) have become a powerful tool for genome editing. Here we present an efficient TALEN assembly approach in which TALENs are assembled by direct Golden Gate ligation into Gateway® Entry vectors from a repeat variable di-residue (RVD) plasmid array. We constructed TALEN pairs targeted to mouse Ddx3 subfamily genes, and demonstrated that our modified TALEN assembly approach efficiently generates accurate TALEN moieties that effectively introduce mutations into target genes. We generated "user friendly" TALEN Entry vectors containing TALEN expression cassettes with fluorescent reporter genes that can be efficiently transferred via Gateway (LR) recombination into different delivery systems. We demonstrated that the TALEN Entry vectors can be easily transferred to an adenoviral delivery system to expand application to cells that are difficult to transfect. Since TALENs work in pairs, we also generated a TALEN Entry vector set that combines a TALEN pair into one PiggyBac transposon-based destination vector. The approach described here can also be modified for construction of TALE transcriptional activators, repressors or other functional domains. © 2013 Zhang et al