Temporal control of gene deletion in sensory ganglia using a tamoxifen-inducible Advillin-Cre-ERT2 recombinase mouse

<p>Abstract</p> <p>Background</p> <p>Tissue-specific gene deletion has proved informative in the analysis of pain pathways. <it>Advillin </it>has been shown to be a pan-neuronal marker of spinal and cranial sensory ganglia. We generated BAC transgenic mice using the <it>Advillin </it>promoter to drive a tamoxifen-inducible CreERT2 recombinase construct in order to be able to delete genes in adult animals. We used a floxed stop <it>ROSA26LacZ </it>reporter mouse to examine functional Cre expression, and analysed the behaviour of mice expressing Cre recombinase.</p> <p>Results</p> <p>We used recombineering to introduce a CreERT2 cassette in place of exon 2 of the <it>Advillin </it>gene into a BAC clone (RPCI23-424F19) containing the 5' region of the <it>Advillin </it>gene. Transgenic mice were generated using pronuclear injection. The resulting <it>AvCreERT2 </it>transgenic mice showed a highly specific expression pattern of Cre activity after tamoxifen induction. Recombinase activity was confined to sensory neurons and no expression was found in other organs. Less than 1% of neurons showed Cre expression in the absence of tamoxifen treatment. Five-day intraperitoneal treatment with tamoxifen (2 mg per day) induced Cre recombination events in ≈90% of neurons in dorsal root and cranial ganglia. Cell counts of dorsal root ganglia (DRG) from transgenic animals with or without tamoxifen treatment showed no neuronal cell loss. Sensory neurons in culture showed ≈70% induction after 3 days treatment with tamoxifen. Behavioural tests showed no differences between wildtype, <it>AvCreERT2 </it>and tamoxifen-treated animals in terms of motor function, responses to light touch and noxious pressure, thermal thresholds as well as responses to inflammatory agents.</p> <p>Conclusions</p> <p>Our results suggest that the inducible pan-DRG <it>AvCreERT2 </it>deleter mouse strain is a useful tool for studying the role of individual genes in adult sensory neuron function. The pain phenotype of the Cre-induced animal is normal; therefore any alterations in pain processing can be unambiguously attributed to loss of the targeted gene.</p

Centrality evolution of the charged-particle pseudorapidity density over a broad pseudorapidity range in Pb-Pb collisions at root s(NN)=2.76TeV

Peer reviewe

(Table 2) Chemical element contents in the surface sediment layer on the submeridional section through the Eastern Pacific

Paper devoted to geochemistry of antimony in metalliferous and transitional sediments from the Southeast Pacific

HDAC4 is required for inflammation-associated thermal hypersensitivity

Transcriptional alterations are characteristic of persistent pain states, but the key regulators remain elusive. HDAC4 is a transcriptional corepressor that has been linked to synaptic plasticity and neuronal excitability, mechanisms that may be involved in peripheral and central sensitization. Using a conditional knockout (cKO) strategy in mice, we sought to determine whether the loss of HDAC4 would have implications for sensory neuron transcription and nociception. HDAC4 was found to be largely unnecessary for transcriptional regulation of naïve sensory neurons but was essential for appropriate transcriptional responses after injury, with Calca and Trpv1 expression consistently down-regulated in HDAC4 cKO compared to levels in the littermate controls (0.2-0.44-fold change, n = 4 in 2 separate experiments). This down-regulation corresponded to reduced sensitivity to 100 nM capsaicin in vitro (IC50 = 230 ± 20 nM, 76 ± 4.4% wild-type capsaicin responders vs. 56.9 ± 4.7% HDAC4 cKO responders) and to reduced thermal hypersensitivity in the complete Freund's adjuvant (CFA) model of inflammatory pain (1.3-1.4-fold improvement over wild-type controls; n = 5-12, in 2 separate experiments). These data indicate that HDAC4 is a novel inflammatory pain mediator and may be a good therapeutic target, capable of orchestrating the regulation of multiple downstream effectors.</p

Työyhteisön suorituskyvyn kehittäminen : case: Tilitoimisto Oy

Tämän opinnäytetyön aiheena on työyhteisön suorituskyvyn kehittäminen tilitoimistossa. Työ keskittyy antamaan kohdeyritykselle keinoja, joiden avulla toimintaa pystyttäisiin kehittämään ja tehostamaan. Opinnäytetyön tavoitteena on selvittää, miten työntekijöiden ja työyhteisön suorituskykyä voidaan kehittää tilitoimiston näkökulmasta ajateltuna. Oppinäytetyössä perehdytään suorituskyvyn muodostumiseen ja sen ylläpitämiseen, sekä tarkastellaan suorituskyvyn vaikutuksia yrityksen kilpailukykyyn ja tuottavuuteen. Työskentely tilitoimistossa on haastavaa jatkuvan kiireen ja määräpäivien vuoksi. Toiminnan laadun turvaamiseksi tilitoimistossa on syytä panostaa myös työntekijöihin työyhteisön toimintaa ja työhyvinvointia kehittämällä. Suorituskykyyn vaikuttavat monet yksilö- ja yhteisötason tekijät. Näiden ollessa tasapainossa voidaan työyhteisössä saavuttaa työhyvinvointia ja näin ollen parantaa työyhteisön tuottavuutta ja tehokkuutta. Työn teoriaosuus on jaettu kolmeen osaan; tilitoimistoalan kuvaaminen, suorituskyvyn muodostuminen ja työhyvinvoinnin vaikutukset suorituskykyyn ja tuottavuuteen. Empiriaosuuden aineisto hankittiin työntekijöille tehdyn kyselyn ja ajankäytön seurannan avulla, lisäksi empiriaosuudessa on tuotu esiin omia havaintojani aiheesta. Empiriaosuuden tulokset osoittavat, että työyhteisön toiminnoissa on kehittämistä. Tuloksista selviää, että työyhteisön tiedonkulussa ja sisäisessä koulutuksessa on aihetta kehittämiseen. Työntekijöiden hyödyntäminen toiminnan suunnittelussa ja kehittämisessä olisi myös tärkeää. Työntekijöiden työaikaa kuluu suurissa osin laskutettaviin tehtäviin, mutta aikavarkaitakin löytyy. Näihin lukeutuvat työn keskeytymiset, keskustelut asiakkaiden kanssa, neuvojen kysyminen toiselta työntekijältä sekä toisaalta toisten työntekijöiden neuvonta. Jatkotutkimuksena voisi selvittää, kuinka hyvin suorituskyvyn ja työyhteisön kehittäminen on vaikuttanut toiminnan tehokkuuteen. Lisäksi voitaisiin verrata työntekijöiden ajankäyttöä sekä todellisuudessa asiakkailta laskutettavia tunteja.The subject of this thesis is the improvement of efficiency and employee performance in an accounting company. The thesis focuses on giving advices to the target company on how develop and improve employee performance. The aim of this thesis is to clarify how the employees’ and the work community’s performance could be developed from accounting company’s point of view. This thesis therefore focuses on the formation of performance and how to maintain it but also on the impacts performance may have on competitiveness and productivity. Working in an accounting company is challenging because of constant rush and deadlines. To secure the quality of performance an accounting company should invest in employees by developing well-being and performance at work. Performance is affected by many factors both at individual level and at the level of the whole work community. When these factors are in balance, the work community may achieve well-being and thereby improve its productivity and efficiency. The theoretical part of the thesis is divided into three parts. First, this section describes the field of accounting. Then, it discusses how performance is formed. Finally, this section examines how well-being affects performance and productivity. The empirical material is collected by employee survey and time tracking but also by my own notices. Based on the results of this study, there is need for improvement in the work community. The results shows that internal communication, training and the overall work climate need improvement. Working time is mainly spent productively, but there are also time stealers such as interruptions, conversations with clients and asking and giving advice to co-workers. Further study could determine, how improving performance and developing the work community have affected productivity. Moreover, employees’ working hours could be compared with the actual chargeable hours

Annotated record of the detailed examination of Mn deposits from DSDP Leg 35 (Holes 322, 323, 324)

The sites drilled on Leg 35 were selected to study the tectonic relationships and interactions between the Antarctic plate and the Antarctic Peninsula, the Scotia Arc, the southern Andes, and the Chile Ridge. Site 322 was drilled on the eastern end of the Bellingshausen Abyssal Plain where it is bounded by the Hero Fracture Zone to the northeast and the continental rise of Antarctica to the south. Site 323 was centrally located on the abyssal plain, about 30 miles west of the Eltanin Fracture Zone and about 150 miles north of the Antarctic continental rise. Site 324 was located on the gently sloping lower continental rise of Antarctica about 160 miles north of Thurston Island, the closest exposed land

Tools for analysis and conditional deletion of subsets of sensory neurons

Background: somatosensation depends on primary sensory neurons of the trigeminal and dorsal root ganglia (DRG). Transcriptional profiling of mouse DRG sensory neurons has defined at least 18 distinct neuronal cell types. Using an advillin promoter, we have generated a transgenic mouse line that only expresses diphtheria toxin A (DTA) in sensory neurons in the presence of Cre recombinase. This has allowed us to ablate specific neuronal subsets within the DRG using a range of established and novel Cre lines that encompass all sets of sensory neurons. Methods: a floxed-tdTomato-stop-DTA bacterial artificial chromosome (BAC) transgenic reporter line (AdvDTA) under the control of the mouse advillin DRG promoter was generated. The line was first validated using a Nav1.8Cre and then crossed to CGRPCreER (Calca), ThCreERT2, Tmem45bCre, Tmem233Cre, Ntng1Cre and TrkBCreER (Ntrk2) lines. Pain behavioural assays included Hargreaves’, hot plate, Randall-Selitto, cold plantar, partial sciatic nerve ligation and formalin tests.Results: motor activity, as assessed by the rotarod test, was normal for all lines tested. Noxious mechanosensation was significantly reduced when either Nav1.8 positive neurons or Tmem45b positive neurons were ablated whilst acute heat pain was unaffected. In contrast, noxious mechanosensation was normal following ablation of CGRP-positive neurons but acute heat pain thresholds were significantly elevated and a reduction in nocifensive responses was observed in the second phase of the formalin test. Ablation of TrkB-positive neurons led to significant deficits in mechanical hypersensitivity in the partial sciatic nerve ligation neuropathic pain model.Conclusions: ablation of specific DRG neuronal subsets using the AdvDTA line will be a useful resource for further functional characterization of somatosensory processing, neuro-immune interactions and chronic pain disorders.<br/