Differential regulation of bladder pain and voiding function by sensory afferent populations revealed by selective optogenetic activation

Bladder-innervating primary sensory neurons mediate reflex-driven bladder function under normal conditions, and contribute to debilitating bladder pain and/or overactivity in pathological states. The goal of this study was to examine the respective roles of defined subtypes of afferent neurons in bladder sensation and function in vivo via direct optogenetic activation. To accomplish this goal, we generated transgenic lines that express a Channelrhodopsin-2-eYFP fusion protein (ChR2-eYFP) in two distinct populations of sensory neurons: TRPV1-lineage neurons (Trpv1Cre;Ai32, the majority of nociceptors) and Nav1.8+ neurons (Scn10aCre;Ai32, nociceptors and some mechanosensitive fibers). In spinal cord, eYFP+ fibers in Trpv1Cre;Ai32 mice were observed predominantly in dorsal horn (DH) laminae I-II, while in Scn10aCre;Ai32 mice they extended throughout the DH, including a dense projection to lamina X. Fiber density correlated with number of retrogradely-labeled eYFP+ dorsal root ganglion neurons (82.2% Scn10aCre;Ai32 vs. 62% Trpv1Cre;Ai32) and degree of DH excitatory synaptic transmission. Photostimulation of peripheral afferent terminals significantly increased visceromotor responses to noxious bladder distension (30–50 mmHg) in both transgenic lines, and to non-noxious distension (20 mmHg) in Scn10aCre;Ai32 mice. Depolarization of ChR2+ afferents in Scn10aCre;Ai32 mice produced low- and high-amplitude bladder contractions respectively in 53% and 27% of stimulation trials, and frequency of high-amplitude contractions increased to 60% after engagement of low threshold (LT) mechanoreceptors by bladder filling. In Trpv1Cre;Ai32 mice, low-amplitude contractions occurred in 27% of trials before bladder filling, which was pre-requisite for light-evoked high-amplitude contractions (observed in 53.3% of trials). Potential explanations for these observations include physiological differences in the thresholds of stimulated fibers and their connectivity to spinal circuits

An S-shaped outflow from IRAS 03256+3055 in NGC 1333

The IRAS source 03256+3055 in the NGC 1333 star forming region is associated with extended sub-millimeter emission of complex morphology, showing multiple clumps. One of these is found to coincide with the driving source of a bipolar jet of S-shaped morphology seen in the emission lines of H_alpha and [SII] as well as in the H2 emission lines in the K-band. Detailed images of the driving source at the wavelengths of H_alpha and [SII] and in the I, J, H, and K bands as well as a K-band spectrum and polarimetry are discussed. The near-infrared morphology is characterized by a combination of line emission from the jet and scattered light from a source with a steep continuum spectrum. The morphology and proper motion of the jet are discussed in the context of a binary system with a precessing disk. We conclude that the molecular core associated with IRAS 03256+3055 consists of several clumps, only one of which shows evidence of recent star formation at optical and near-infrared wavelengths.We also briefly discuss a second, newly found near-infrared source associated with a compact sub-millimeter continuum source near IRAS 03256+3055, and conclude that this source may be physically unrelated the cluster of molecular clumps.Comment: 25 pages, including 5 figures. Accepted for publication in The Astronomical Journa

Loss of the PTCH1 tumor suppressor defines a new subset of plexiform fibromyxoma.

BackgroundPlexiform fibromyxoma (PF) is a rare gastric tumor often confused with gastrointestinal stromal tumor. These so-called "benign" tumors often present with upper GI bleeding and gastric outlet obstruction. It was recently demonstrated that approximately one-third of PF have activation of the GLI1 oncogene, a transcription factor in the hedgehog (Hh) pathway, via a MALAT1-GLI1 fusion protein or GLI1 up-regulation. Despite this discovery, the biology of most PFs remains unknown.MethodsNext generation sequencing (NGS) was performed on formalin-fixed paraffin-embedded (FFPE) samples of PF specimens collected from three institutions (UCSD, NCI and OHSU). Fresh frozen tissue from one tumor was utilized for in vitro assays, including quantitative RT-PCR and cell viability assays following drug treatment.ResultsEight patients with PF were identified and 5 patients' tumors were analyzed by NGS. An index case had a mono-allelic PTCH1 deletion of exons 15-24 and a second case, identified in a validation cohort, also had a PTCH1 gene loss associated with a suspected long-range chromosome 9 deletion. Building on the role of Hh signaling in PF, PTCH1, a tumor suppressor protein, functions upstream of GLI1. Loss of PTCH1 induces GLI1 activation and downstream gene transcription. Utilizing fresh tissue from the index PF case, RT-qPCR analysis demonstrated expression of Hh pathway components, SMO and GLI1, as well as GLI1 transcriptional targets, CCND1 and HHIP. In turn, short-term in vitro treatment with a Hh pathway inhibitor, sonidegib, resulted in dose-dependent cell killing.ConclusionsFor the first time, we report a novel association between PTCH1 inactivation and the development of plexiform fibromyxoma. Hh pathway inhibition with SMO antagonists may represent a target to study for treating a subset of plexiform fibromyxomas

2002. Using two-photon standing waves and patterned photobleaching to measure diffusion from nanometers to microns in biological systems

A method of measuring molecular diffusion rates in microscopic sample volumes is described. This method utilizes the standing wave interference created by colliding two counterpropagating laser beams at the focus of two opposing microscope objectives, creating a periodic light distribution in a volume on the order of 1 fl. By using a Pockels cell to vary the laser intensity with a time resolution of milliseconds, we show how this experimental geometry can be used to perform ultrahigh resolution fluorescence recovery after patterned photobleaching ͑FRAPP͒ experiments. A mathematical treatment of the experiment shows that the laser excitation profile has two characteristic length scales, the width of the focal spot and the period of the standing wave, which permits the simultaneous measurement of dynamics on two separate length scales. This feature may be used to determine whether the measured diffusion is anomalous. We present experimental results using a femtosecond Ti:sapphire laser to create a two-photon excitation profile with a fringe visibility on the order of 100. This standing wave is used to demonstrate FRAPP in both model dye/polymer systems and in more complex systems like living cells stained with a fluorescent dye. By combining the advantages of standing wave microscopy and two-photon fluorescence recovery after photobleaching, this technique permits the measurement of very short length motions in localized sample volumes, which should be useful in both biology and the study of diffusion in microscopically heterogeneous systems

Growth dynamics of a Bose-Einstein condensate in a dimple trap without cooling

We study the formation of a Bose-Einstein condensate in a cigar-shaped three-dimensional harmonic trap, induced by the controlled addition of an attractive "dimple" potential along the weak axis. In this manner we are able to induce condensation without cooling due to a localized increase in the phase space density. We perform a quantitative analysis of the thermodynamic transformation in both the sudden and adiabatic regimes for a range of dimple widths and depths. We find good agreement with equilibrium calculations based on self-consistent semiclassical Hartree-Fock theory describing the condensate and thermal cloud. We observe there is an optimal dimple depth that results in a maximum in the condensate fraction. We also study the non-equilibrium dynamics of condensate formation in the sudden turn-on regime, finding good agreement for the observed time dependence of the condensate fraction with calculations based on quantum kinetic theory.Comment: v1: 9 pages, 7 figures, submitted to Phys. Rev. A; v2: 10 pages, 8 figures, fixed typos, added references, additional details on experimental procedure, values of phase-space density, new figure and discussion on effects of three-body loss in Appendix B (replaced with published version

Shearographic non-destructive evaluation of space shuttle thermal protection systems

Preliminary results of shearographic inspections of the shuttle external tank (ET) spray-on foam insulation (SOFI) and solid rocket booster (SRB) Marshall sprayable ablative (MSA-2) epoxy-cork thermal protection systems (TPS) are presented. Debonding SOFI or MSA-2 damage the orbiter 'belly' tile and exposes the ET/SRB to thermal loading. Previous work with the ET/SRB showed promising results with shearography. The first area investigated was the jack pad close-out, one of many areas on the ET where foam is applied at KSC. Voids 0.375 inch were detected in 1.75 inch thick foam using a pressure reduction of less than 0.4 psi. Of primary interest are areas of the ET that directly face the orbiter tile TPS. It is estimated that 90% of tile TPS damage on the orbiter 'belly' results from debonding SOFI during ascent. Test panels modeling these areas were manufactured with programmed debonds to determine the sensitivity of shearography as a function of debond size, SOFI thickness and vacuum. Results show repeatable detection of debonds with a diameter approximately half the SOFI thickness at less than 0.4 psi pressure reduction. Preliminary results are also presented on inspections of MSA-2 and the remote manipulator system (RMS) honeycomb materia

CSU radiation budget pilot study for TOGA COARE

July 1993.Funding agencies: National Oceanic and Atmospheric Administration, Office of Naval Research

Comparative landscape dynamics of two anuran species: climate‐driven interaction of local and regional processes

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/117162/1/ecm2009793503.pd