OB Stars in the Solar Neighborhood I: Analysis of their Spatial Distribution

We present a newly-developed, three-dimensional spatial classification method, designed to analyze the spatial distribution of early type stars within the 1 kpc sphere around the Sun. We propose a distribution model formed by two intersecting disks -the Gould Belt (GB) and the Local Galactic Disk (LGD)- defined by their fundamental geometric parameters. Then, using a sample of about 550 stars of spectral types earlier than B6 and luminosity classes between III and V, with precise photometric distances of less than 1 kpc, we estimate for some spectral groups the parameters of our model, as well as single membership probabilities of GB and LGD stars, thus drawing a picture of the spatial distribution of young stars in the vicinity of the Sun.Comment: 28 pages including 9 Postscript figures, one of them in color. Accepted for publication in The Astronomical Journal, 30 January 200

On Scaling Laws and Alfvenic Magnetic Fluctuations in Molecular Clouds

Under the basic assumption that the observed turbulent motions in molecular clouds are Alfvenic waves or turbulence, we emphasize that the Doppler broadening of molecular line profiles directly measures the velocity amplitudes of the waves instead of the Alfven velocity. Assuming an equipartition between the kinetic energy and the Alfvenic magnetic energy, we further propose the hypothesis that observed standard scaling laws in molecular clouds imply a roughly scale-independent fluctuating magnetic field, which might be understood as a result of strong wave-wave interactions and subsequent energy cascade. We predict that $\sigma_{v}\propto \rho^{-0.5}$ is a more basic and robust relation in that it may approximately hold in any regions where the spatial energy density distribution is primarily determined by wave-wave interactions, including gravitationally unbound regions. We also discuss the fact that a scale-independent $\sigma_{B}^{2}$ appears to contradict existing 1-D and 2-D computer simulations of MHD turbulence in molecular clouds.Comment: 11 pages, to appear in Astrophysical Journal Letters, email: [email protected]

Resident Cardiac Immune Cells and Expression of the Ectonucleotidase Enzymes CD39 and CD73 after Ischemic Injury

BACKGROUND: The ectoenzymes CD39 and CD73 are expressed by a broad range of immune cells and promote the extracellular degradation of nucleotides to anti-inflammatory adenosine. This study explored the abundance of CD73 and CD39 on circulating and resident cardiac leukocytes and coronary endothelial cells under control conditions and in response to inflammation following myocardial ischemia and reperfusion (I/R). METHODS AND RESULTS: A method was elaborated to permit FACS analysis of non-myocardial cells (resident leukocytes, coronary endothelium and CD31(-) CD45(-) cells) of the unstressed heart. Under control conditions the murine heart contained 2.3 × 10(3) resident leukocytes/mg tissue, the most prominent fraction being antigen-presenting mononuclear cells (CD11b(+) CD11c(+) F4/80(+) MHCII(+)) followed by B-cells, monocytes and T-cells. CD73 was highly expressed on circulating and resident cardiac lymphoid cells with little expression on myeloid cells, while the opposite was true for CD39. Cardiomyocytes and erythrocytes do not measurably express CD39/CD73 and CD39 dominates on coronary endothelium. Three days after I/R, CD73 was significantly upregulated on invading granulocytes (2.8-fold) and T-cells (1.5-fold). Compared with coronary endothelial cells, CD73 associated with leukocytes comprised 2/3 of the total cardiac CD73. CONCLUSION: Our study suggests that extracellular ATP formed during I/R is preferentially degraded by CD39 present on myeloid cells, while the formation of immunosuppressive adenosine is mainly catalysed by CD73 present on granulocytes and lymphoid cells. Upregulated CD73 on granulocytes and T-cells infiltrating the injured heart is consistent with the existence of an autocrine adenosinergic loop which may promote the healing process

Remodeling of Purinergic Receptor-Mediated Ca2+ Signaling as a Consequence of EGF-Induced Epithelial-Mesenchymal Transition in Breast Cancer Cells

Background The microenvironment plays a pivotal role in tumor cell proliferation, survival and migration. Invasive cancer cells face a new set of environmental challenges as they breach the basement membrane and colonize distant organs during the process of metastasis. Phenotypic switching, such as that which occurs during epithelial-mesenchymal transition (EMT), may be associated with a remodeling of cell surface receptors and thus altered responses to signals from the tumor microenvironment. Methodology/Principal Findings We assessed changes in intracellular Ca 2+ in cells loaded with Fluo-4 AM using a fluorometric imaging plate reader (FLIPR TETRA) and observed significant changes in the potency of ATP (EC 50 0.175 μM (-EGF) versus 1.731 μM (+EGF), P<0.05), and the nature of the ATP-induced Ca 2+ transient, corresponding with a 10-fold increase in the mesenchymal marker vimentin (P<0.05). We observed no change in the sensitivity to PAR2-mediated Ca 2+ signaling, indicating that these alterations are not simply a consequence of changes in global Ca 2+ homeostasis. To determine whether changes in ATP-mediated Ca 2+ signaling are preceded by alterations in the transcriptional profile of purinergic receptors, we analyzed the expression of a panel of P2X ionotropic and P2Y metabotropic purinergic receptors using real-time RT-PCR and found significant and specific alterations in the suite of ATP-activated purinergic receptors during EGF-induced EMT in breast cancer cells. Our studies are the first to show that P2X 5 ionotropic receptors are enriched in the mesenchymal phenotype and that silencing of P2X 5 leads to a significant reduction (25%, P<0.05) in EGF-induced vimentin protein expression. Conclusions The acquisition of a new suite of cell surface purinergic receptors is a feature of EGF-mediated EMT in MDA-MB-468 breast cancer cells. Such changes may impart advantageous phenotypic traits and represent a novel mechanism for the targeting of cancer metastasis

In pursuit of P2X3 antagonists: novel therapeutics for chronic pain and afferent sensitization

Treating pain by inhibiting ATP activation of P2X3-containing receptors heralds an exciting new approach to pain management, and Afferent's program marks the vanguard in a new class of drugs poised to explore this approach to meet the significant unmet needs in pain management. P2X3 receptor subunits are expressed predominately and selectively in so-called C- and Aδ-fiber primary afferent neurons in most tissues and organ systems, including skin, joints, and hollow organs, suggesting a high degree of specificity to the pain sensing system in the human body. P2X3 antagonists block the activation of these fibers by ATP and stand to offer an alternative approach to the management of pain and discomfort. In addition, P2X3 is expressed pre-synaptically at central terminals of C-fiber afferent neurons, where ATP further sensitizes transmission of painful signals. As a result of the selectivity of the expression of P2X3, there is a lower likelihood of adverse effects in the brain, gastrointestinal, or cardiovascular tissues, effects which remain limiting factors for many existing pain therapeutics. In the periphery, ATP (the factor that triggers P2X3 receptor activation) can be released from various cells as a result of tissue inflammation, injury or stress, as well as visceral organ distension, and stimulate these local nociceptors. The P2X3 receptor rationale has aroused a formidable level of investigation producing many reports that clarify the potential role of ATP as a pain mediator, in chronic sensitized states in particular, and has piqued the interest of pharmaceutical companies. P2X receptor-mediated afferent activation has been implicated in inflammatory, visceral, and neuropathic pain states, as well as in airways hyperreactivity, migraine, itch, and cancer pain. It is well appreciated that oftentimes new mechanisms translate poorly from models into clinical efficacy and effectiveness; however, the breadth of activity seen from P2X3 inhibition in models offers a realistic chance that this novel mechanism to inhibit afferent nerve sensitization may find its place in the sun and bring some merciful relief to the torment of persistent discomfort and pain. The development philosophy at Afferent is to conduct proof of concept patient studies and best identify target patient groups that may benefit from this new intervention

Continuous-Flow Preparation and Use of β‑Chloro Enals Using the Vilsmeier Reagent

The Vilsmeier reagent is used in the preparation of a wide variety of heterocycles, such as pyrazoles, <i>via</i> formation of β-chloroacrolein intermediates. However, use of this extremely reactive reagent on large scale requires special precautions to avoid potentially dangerous exotherms. This article describes the safe preparation at room temperature of the Vilsmeier reagent under flow conditions for the formation of β-chloroacroleins and 3-formylchromones, as well as the use of these in multistep, continuous flow processes for the syntheses of β-acrylonitriles and polysubstituted pyrazoles

Real time detection of extracellular ATP in tumor microenvironment.

Background: The tumor microenvironment plays a critical role in tumor initiation, progression and invasion. Nucleotides, such as ATP, have a surprisingly wide range of modulatory effects on tumor cells, however techniques for measuring ATP in the extracellular environment are still rudimentary. Here we show an in vivo method for measuring extracellular ATP release in tumor microenvironment. Methods: We stably expressed pmeLUC, a chimeric luciferase targeted to the extracellular side of the plasma membrane, in HEK293 cell lines. A stable cell clone (HEK293-pmeLUC) was then injected anesthetized nude/nude mice bearing or not a tumor. Bioluminescence was then monitored by IVIS imaging system (Xenogen). Results: Endovenous, intraperitoneal and subcutaneous inoculation of HEK293-pmeLUC cells in healthy nude mice caused a modest, barely detectable, increase in luminescence. On the contrary, bioluminescence observed after injection of the probe into mice bearing the OVCAR-3 human ovarian carcinoma or the MZ2-MEL human melanoma increased strongly. We then validated HEK293-pmeLUC cells as ATP reporters using the ATP-hydrolyzing enzyme, potato apyrase. Apyrase injection cause a significative drop in luminescence. We then performed an in vitro calibration which revealed that ATP in the tumor interstitium is in the hundrend micromolar range, while it is basically undetectable in healthy tissues. Our approach offers a new tool for the investigation of the biochemical composition of tumor milieu and for development of novel therapies based on the modulation of extracellular purine-based signalling