Yin Yang of immunoregulation in organ transplantation and cancer

The ‘Yin Yang of Immunoregulation: Cancer and Organ Transplantation’ meeting took place in Nantes, France on 2–3 December 2010 and was dedicated to the biology of myeloid and lymphoid immune cells in the context of cancer and transplantation. This meeting was organized by the Immunotherapy Research group of the Western France Cancer Network Cancéropole Grand-Ouest and the Immunomonitorage et Biothérapies network (IMBIO) research program, which is supported by the Région Pays de la Loire

How interdisciplinary is nanotechnology?

Facilitating cross-disciplinary research has attracted much attention in recent years, with special concerns in nanoscience and nanotechnology. Although policy discourse has emphasized that nanotechnology is substantively integrative, some analysts have countered that it is really a loose amalgam of relatively traditional pockets of physics, chemistry, and other disciplines that interrelate only weakly. We are developing empirical measures to gauge and visualize the extent and nature of interdisciplinary interchange. Such results speak to research organization, funding, and mechanisms to bolster knowledge transfer. In this study, we address the nature of cross-disciplinary linkages using “science overlay maps” of articles, and their references, that have been categorized into subject categories. We find signs that the rate of increase in nano research is slowing, and that its composition is changing (for one, increasing chemistry-related activity). Our results suggest that nanotechnology research encompasses multiple disciplines that draw knowledge from disciplinarily diverse knowledge sources. Nano research is highly, and increasingly, integrative—but so is much of science these days. Tabulating and mapping nano research activity show a dominant core in materials sciences, broadly defined. Additional analyses and maps show that nano research draws extensively upon knowledge presented in other areas; it is not constricted within narrow silos

Codon swapping of zinc finger nucleases confers expression in primary cells and in vivo from a single lentiviral vector

BACKGROUND: Zinc finger nucleases (ZFNs) are promising tools for genome editing for biotechnological as well as therapeutic purposes. Delivery remains a major issue impeding targeted genome modification. Lentiviral vectors are highly efficient for delivering transgenes into cell lines, primary cells and into organs, such as the liver. However, the reverse transcription of lentiviral vectors leads to recombination of homologous sequences, as found between and within ZFN monomers. METHODS: We used a codon swapping strategy to both drastically disrupt sequence identity between ZFN monomers and to reduce sequence repeats within a monomer sequence. We constructed lentiviral vectors encoding codon-swapped ZFNs or unmodified ZFNs from a single mRNA transcript. Cell lines, primary hepatocytes and newborn rats were used to evaluate the efficacy of integrative-competent (ICLV) and integrative-deficient (IDLV) lentiviral vectors to deliver ZFNs into target cells. RESULTS: We reduced total identity between ZFN monomers from 90.9% to 61.4% and showed that a single ICLV allowed efficient expression of functional ZFNs targeting the rat UGT1A1 gene after codon-swapping, leading to much higher ZFN activity in cell lines (up to 7-fold increase compared to unmodified ZFNs and 60% activity in C6 cells), as compared to plasmid transfection or a single ICLV encoding unmodified ZFN monomers. Off-target analysis located several active sites for the 5-finger UGT1A1-ZFNs. Furthermore, we reported for the first time successful ZFN-induced targeted DNA double-strand breaks in primary cells (hepatocytes) and in vivo (liver) after delivery of a single IDLV encoding two ZFNs. CONCLUSION: These results demonstrate that a codon-swapping approach allowed a single lentiviral vector to efficiently express ZFNs and should stimulate the use of this viral platform for ZFN-mediated genome editing of primary cells, for both ex vivo or in vivo applications

Lentiviral Vectors That Express UGT1A1 in Liver and Contain Mir-142 Target Sequences Normalize Hyperbilirubinemia in Gunn Rats

Background & AimsCrigler–Najjar type 1 (CN-I) is an inherited liver disease caused by an absence of bilirubin–uridine 5′-diphosphate–glucuronosyltransferase (UGT1A1) activity. It results in life-threatening levels of unconjugated bilirubin, and therapeutic options are limited. We used adult Gunn rats (an animal model of the disease) to evaluate the efficiency of lentiviral-based gene therapy to express UGT1A1 in liver. Methods Gunn rats were given intraportal injections of VSVG-pseudotyped lentiviral vectors that encode UGT1A1 under the control of a liver-specific transthyretin promoter (mTTR.hUGT1A1); this vector does not contain target sequences for miR-142, a microRNA that is expressed specifically in hematopoietic cells. Rats were also injected with the vector mTTR.hUGT1A1.142T, which contains 4 copies of the miR-142 target sequences; its messenger RNA should be degraded in antigen-presenting cells. Bilirubinemia was monitored, and the presence of transduced hepatocytes was analyzed by quantitative polymerase chain reaction. Vector expression was tested in vitro in rat hematopoietic cells. Results In Gunn rats, bilirubin levels normalized 2 weeks after administration of mTTR.hUGT1A1. However, hyperbilirubinemia resumed 8 weeks after vector administration, concomitant with the induction of an immune response. In contrast, in rats injected with mTTR-UGT1A1.142T, bilirubin levels normalized for up to 6 months and transduced cells were not eliminated. Conclusions Lentiviral vectors that express UGT1A1 reduce hyperbilirubinemia in immunocompetent Gunn rats for at least 6 months. The immune response against virally expressed UGT1A1 can be circumvented by inclusion of miR-142 target sequences, which reduce vector expression in antigen-presenting cells. This lentiviral-based gene therapy approach might be developed to treat patients with CN-I

Méganucléases, Zinc-finger nucléases et autres nucléases... de nouveaux outils pour la Transgenèse. Présentation de ces nouveaux outils, intérêts et perspectives. Exploration fonctionnelle.

Communication oral

Ligand binding to Fc gamma R induces c-myc-dependent apoptosis in IL-2-stimulated NK cells.

The role of signals transduced via Fc gamma RIIIA in the modulation of the proliferative potential of NK cells has been investigated. Fc gamma R stimulation does not induce NK cell proliferation, and inhibits that induced by IL-2, but not by IL-12, as measured by [3H]TdR incorporation, without affecting entrance or progression through cell cycle. The inhibitory effect depends, at least in part, on induced apoptosis of the cells, detected by both light and electron microscopy examination. Fc gamma R stimulation induces apoptosis only in NK cells that have been previously activated by IL-2: this occurs within 3 h from receptor stimulation and is independent from de novo receptor-induced RNA or protein synthesis, but requires receptor-induced activation of protein tyrosine kinases and extracellular Ca2+ influx. IL-2 induces accumulation of c-myc mRNA in NK cells, and treatment of the cells with c-myc antisense oligodeoxyribonucleotides during the IL-2 stimulation phase inhibits the susceptibility to Fc gamma RIIIA-induced cell death, indicating that the induction of sustained levels of this proto-oncogene is necessary for the phenomenon. Thus, a two-step model is suggested for the Fc gamma R-induced apoptosis in IL-2 activated NK cells: the first step involves induced expression of c-myc, and possibly other permissive factors, upon IL-2 prestimulation; the second depends directly on the stimulation of the receptor, independently of additional gene induction. The evidence presented here suggests a mechanism of control of NK cell expansion at the latest stages of Ab-dependent immune responses

Customer experience in multimodal food service in pop up -restaurant : Meaning of space, product, encounter and atmosphere for customer experience

Elämys määritetään kokonaisvaltaiseksi, moniaistiseksi ja yksilölliseksi kokemukseksi, joka jättää muistijäljen. Siihen sisältyy myönteisiä tunteita, aktiivisuutta ja syvää tiedostamista sekä keskittymistä. Elämyskulutus voidaan nähdä symbolikulutuksena, jonka tarkoituksena on viestiä identiteetistä, sosiaalisesta statuksesta ja kuulumisesta tiettyyn ryhmään. Yksilöt tavoittelevat elämyksiä myös tehdäkseen elämästään merkityksellisemmän. Niinpä 1990-luvulla elämykset on nostettu vahvasti sekä yleiseen että akateemiseen keskusteluun määrittämällä ne selvästi uusiksi taloudellisiksi mahdollisuuksiksi, jotka ovat tuotteistettavissa. Elämysten tuottamisen mahdollisuudet kilpailukykytekijänä on havaittu myös muualla kuin matkailualalla. Ravintolapalvelujakin on ryhdytty kehittämään elämyksellisempään suuntaan ja kiinnostus pop up -tyyppisesti tuotettuja palveluja kohtaan on lisääntynyt. Ravintolapalvelun onnistuneisuutta määrittelee pitkälle asiakkaiden kokonaiskokemus. Sillä on merkittävä vaikutus muun muassa asiakkaiden haluun palata palveluun. Asiakaskokemus heijastuu myös asiakkaiden palvelussa kuluttaman rahan määrään, haluun kertoa myönteistä viestiä kokemuksesta sekä haluun sitoutua pysyvästi palvelun käyttäjäksi. Tämän tutkimuksen tarkoitus on löytää tekijät, joilla on merkitystä ravintolan asiakkaan kokemuksen parantamisessa. Tutkimuksen tärkein tavoite on kuvata sitä, mitä havaintoja asiakkaat tekivät aterian aikana ja minkälainen on asiakkaille syntynyt kokonaiskokemus tapahtumasta. Tutkimuksessa pyritään myös kartoittamaan ne tekijät, jotka aiheuttavat voimakkaimpia myönteisiä ja kielteisiä kokemuksia. Tutkimusaineisto on kerätty Juurella-ravintolan toteuttamien elämyksellisten pop up -ravintolaillallisten yhteydessä asiakkailta välittömästi kokemuksen jälkeen. Tutkimusaineiston keräämiseen ja asiakaskokemusten arvioimiseen käytettiin teatterimaista tutkimustapaa, ns. asiakaskriittistä lähestymistä. Tehtävään rekrytoitiin kolme fokusryhmää, joissa kussakin oli kuusi henkilöä, miehiä ja naisia iältään 18 – 63 vuotiaita. Pop up -ravintolatapahtumat järjestettiin kolmella paikkakunnalla, Kurikassa, Jalasjärvellä ja Alavudella tyypillisistä ravintoloista poikkeavissa tiloissa. Tulokset korostavat tunnelman moniaistisia piirteitä. Subjektiiviset ja kulttuuriset tulkinnat ovat aina mukana aistimuksissamme. Tulosten perusteella voidaan sanoa, että ihmiset antavat henkilökohtaisia merkityksiä kokemuksilleen ja tekevät assosiaatioita eri kokemuselementtien välillä. Voidaan sanoa, että ihmiset havaitsevat pikkutarkasti eri asioita palvelukokonaisuudessa. Aistiärsykkeiden voimakkuus ateriointitilanteessa vaikuttaa vahvasti siihen, minkälainen on havaintojen keskinäinen painotus ja minkälaiseksi kokonaiskokemus muodostuu. Uudet kokemukset tuottavat vahvoja mielikuvia ja provosoivat keskustelua sekä pöytäseurueen että asiakaspalvelijoiden kanssa. Havahduttavat asiat aktivoivat ja ne huomataan herkimmin. Esimerkiksi Jalasjärvellä eniten keskustelua herätti poikkeuksellinen pellavatehtaan tila ja Alavudella puolestaan itse ruokakokemus. Nostalgiakokemukset nostavat herkästi muistoja mieleen ja myönteiset muistot vahvistavat yhteisöllisyyttä ja yhteisen kokemuksen tunnetta. Paikat ja tilat kaikissa kolmessa kohteessa loivat moniaistisia ja nostalgisia kokemuksia, joita arvostettiin paljon. Asiakaskokemukset olivat voittopuolisesti hyvin myönteisiä, mutta muutama kielteinenkin havainto todettiin.Extraordinary experience is an overall, multimodal and individual experience that leaves an engram. Positive emotions, activity, deep awareness and concentration are included. Experience consumption may be seen as symbolic consumption that tries to communicate identity, social status and belong to certain group. Individuals chase after experiences in order to make their life more meaningful. During 1990`s general and academic discussion about experiences have been taken out and they were clearly found new economic opportunities that can be commercialized. These opportunities were noticed as competitive power also outside tourism. Restaurant services have also developed to more experiential direction and interest in pop up -restaurants has increased. The holistic customer experience strongly defines the success of restaurant service. It has an effect on customer`s willingness to come back. Customer experience has also an impact on the consumed amount of money, the willingness to use positive word of mouth and the willingness to show customer loyalty. Purpose of this study is to find factors that have meaning in improving customer experience. The most important goal is to describe what kind of perceptions customers made during the dinner and what kind is the holistic customer experience. The factors that cause strongest positive and negative experiences are also described. The data is collected right after customers had experienced their dinners in three experiential pop up -events organized by Restaurant Juurella. Theatrical method, so called customer critic approach, is used as tool in collecting the data. Three focus groups were recruited, six persons in each group, men and women ages 18 to 63. Experiential pop up -events were organized in three towns, Kurikka, Jalasjärvi, Alavus, in unexceptional places. Result highlight the multisensory aspects of atmosphere. Subjective and cultural interpretations are always included in sensing. According to the results it can be mentioned that people give personal meanings to their experiences and make associations between different experience elements. People also make detailed perceptions in all different service elements. Strength in sensory stimulus have effect on what is the emphasis between different perceptions and what the overall experience will consist of. New experiences produce strong images and provoke discussion with both party around the table and personnel. Arousing things will activate and they will be perceived easier. For example in Jalasjärvi linen factory guests spoke most about the space and in Alavus they discussed most about the food experience. Nostalgic experiences raise delicately up memories and positive memories make sense of community and common experience stronger. Places and spaces in every three event created multimodal and nostalgic experiences that were highly appreciated. Customer experiences were predominantly very positive but some negative opinions were also found