Effects of P300-based BCI use on reported presence in a virtual environment

Brain-computer interfaces (BCIs) are becoming more and more popular as an input device for virtual worlds and computer games. Depending on their function, a major drawback is the mental workload associated with their use and there is significant effort and training required to effectively control them. In this paper, we present two studies assessing how mental workload of a P300-based BCI affects participants" reported sense of presence in a virtual environment (VE). In the first study, we employ a BCI exploiting the P300 event-related potential (ERP) that allows control of over 200 items in a virtual apartment. In the second study, the BCI is replaced by a gaze-based selection method coupled with wand navigation. In both studies, overall performance is measured and individual presence scores are assessed by means of a short questionnaire. The results suggest that there is no immediate benefit for visualizing events in the VE triggered by the BCI and that no learning about the layout of the virtual space takes place. In order to alleviate this, we propose that future P300-based BCIs in VR are set up so as require users to make some inference about the virtual space so that they become aware of it,which is likely to lead to higher reported presence.

Data communication in paper sales environment

Tämä diplomityö kirjoitettiin UPM- Kymmene konsernin UPM Net Services sa/nv osastolle Brysselissä ja Helsingissä. Työn aihe, Data communication in paper sales environment, määriteltiin käsittelemään paperin myyntijärjestelmään liittyviä aiheita. Nykyinen paperin myyntijärjestelmä on käsitelty ensin teoriassa ja aiheeseen kuuluvat ohjelmistotuotteet ja työkaluohjelmistot on esitelty. Parannuksia nykyiseen järjestelmään on pohdittu ohjelmistosuunnittelun, tehokkuuden, tiedon hallinnan, tietoturvallisuuden ja liiketoiminnan näkökulmista. Diplomityön käytännön osuudessa esitellään kaksi ohjelmistoa. Nämä ohjelmistot tehtiin UPM Net Services'lle, jotta saatiin kokemuksia viestin välitykseen perustuvasta tiedon siirrosta. Diplomityön johtopäätösosuudessa todetaan, että paperin myyntijärjestelmän tiedon siirto toimii luotettavasti nykyisessä järjestelmässä. Tulevaisuuden tarpeet ja parannukset ovat kuitenkin vaikeasti toteutettavissa nykyään käytettävin välinein. Erityisesti internetin hyödyntäminen nähdään tärkeänä, mutta se on vaikeasti otettavissa käyttöön nykyisessä järjestelmässä. Viestin välitykseen perustuvat järjestelmät ovat osoittautuneet käytännössä toimiviksi ja tärkein kehitysehdotus onkin viestin välitysjärjestelmän käyttöönotto.

Water consumption and soil moisture distribution in melon crop with mulching and in a protected environment

Mulching has become an important technique for land cover, but there are some technical procedures which should be adjusted for these new modified conditions to establish optimum total water depth. It is also important to observe the soil-water relations as soil water distribution and wetted volume dimensions. The objective of the present study was to estimate melon evapotranspiration under mulching in a protected environment and to verify the water spatial distribution around the melon root system in two soil classes. Mulching provided 27 mm water saving by reducing water evaporation. In terms of volume each plant received, on average, the amount of 175.2 L of water in 84 days of cultivation without mulching, while when was used mulching the water requirement was 160.2 L per plant. The use of mulching reduced the soil moisture variability throughout the crop cycle and allowed a greater distribution of soil water that was more intense in the clay soil. The clayey soil provided on average 43 mm more water depth retention in 0.50 m soil deep relative to the sandy loam soil, and reduced 5.6 mm the crop cycle soil moisture variation compared to sandy loam soil.

The political economy of international regulatory convergence in public utilities

To what extent should public utilities regulation be expected to converge across countries? When it occurs, will it generate good outcomes? Building on the core proposition of the New Institutional Economics that similar regulations generate different outcomes depending on their fit with the underlying domestic institutions, we develop a simple model and explore its implications by examining the diffusion of local loop unbundling (LLU) regulations. We argue that: one should expect some convergence in public utility regulation but with still a significant degree of local experimentation; this process will have very different impacts of regulation.

Political market and regulatory uncertainty: Insights and implications for integrated strategy

Managers can craft effective integrated strategy by properly assessing regulatory uncertainty. Leveraging the existing political markets literature, we predict regulatory uncertainty from the novel interaction of demand and supply side rivalries across a range of political markets. We argue for two primary drivers of regulatory uncertainty: ideology-motivated interests opposed to the firm and a lack of competition for power among political actors supplying public policy. We align three, previously disparate dimensions of nonmarket strategy - profile level, coalition breadth, and pivotal target - to levels of regulatory uncertainty. Through this framework, we demonstrate how and when firms employ different nonmarket strategies. To illustrate variation in nonmarket strategy across levels of regulatory uncertainty, we analyze several market entry decisions of foreign firms operating in the global telecommunications sector.

Sterol regulatory element binding protein-1a transactivates 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase gene promoter

6-Phosphofructo-2-kinase/fructose-2,6-bisphosphatase (PFKFB)catalyzes the synthesis and degradation of fructose-2,6-bisphosphate, a key modulator of glycolysis-gluconeogenesis. To gain insight into the molecular mechanism behind hormonal and nutritional regulation of PFKFB expression, we have cloned and characterized the proximal promoter region of the liver isoform of PFKFB (PFKFB1) from gilthead sea bream (Sparus aurata). Transient transfection of HepG2 cells with deleted gene promoter constructs and electrophoretic mobility shift assays allowed us to identify a sterol regulatory element (SRE) to which SRE binding protein-1a (SREBP-1a)binds and transactivates PFKFB1 gene transcription. Mutating the SRE box abolished SREBP-1a binding and transactivation. The in vivo binding of SREBP-1a to the SRE box in the S. aurata PFKFB1 promoter was confirmed by chromatin immunoprecipitation assays. There is a great deal of evidence for a postprandial rise of PFKB1 mRNA levels in fish and rats. Consistently, starved-to-fed transition and treatment with glucose or insulin increased SREBP-1 immunodetectable levels, SREBP-1 association to PFKFB1 promoter, and PFKFB1 mRNA levels in the piscine liver. Our findings demonstrate involvement of SREBP-1a in the transcriptional activation of PFKFB1, and we conclude that SREBP-1a may exert a key role mediating postprandial activation of PFKFB1 transcription.

STACK OVERWRITING ATTACKS AND DEFENCES IN UNIX ENVIRONMENT

Työn tarkoituksena on tutkia pinon ylikirjoitukseen perustuvien hyökkäysten toimintaa ja osoittaa kokeellisesti nykyisten suojaustekniikoiden olevan riittämättömiä. Tutkimus suoritetaan testaamalla miten valitut tietoturvatuotteet toimivat eri testitilanteissa. Testatut tuotteet ovat Openwall, PaX, Libsafe 2.0 ja Immunix 6.2. Testaus suoritetaan pääasiassa RedHat 7.0 ympäristössä testiohjelman avulla. Testeissä mitataan sekä tuotteiden kyky havaita hyökkäyksiä että niiden nopeusvaikutukset. Myös erityyppisten hyökkäysten ja niitä vastaan kehitettyjen metodien toimintaperiaatteet esitellään seikkaperäisesti ja havainnollistetaan yksinkertaistetuilla esimerkeillä. Esitellyt tekniikat sisältävät puskurin ylivuodot, laittomat muotoiluparametrit, loppumerkittömät merkkijonot ja taulukoiden ylivuodot. Testit osoittavat, etteivät valitut tuotteet estä kaikkia hyökkäyksiä, joten lopuksi perehdytään myös vahinkojen minimointiin onnistuneiden hyökkäysten varalta.

Sterol regulatory element binding protein-1a transactivates 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase gene promoter

6-Phosphofructo-2-kinase/fructose-2,6-bisphosphatase (PFKFB)catalyzes the synthesis and degradation of fructose-2,6-bisphosphate, a key modulator of glycolysis-gluconeogenesis. To gain insight into the molecular mechanism behind hormonal and nutritional regulation of PFKFB expression, we have cloned and characterized the proximal promoter region of the liver isoform of PFKFB (PFKFB1) from gilthead sea bream (Sparus aurata). Transient transfection of HepG2 cells with deleted gene promoter constructs and electrophoretic mobility shift assays allowed us to identify a sterol regulatory element (SRE) to which SRE binding protein-1a (SREBP-1a)binds and transactivates PFKFB1 gene transcription. Mutating the SRE box abolished SREBP-1a binding and transactivation. The in vivo binding of SREBP-1a to the SRE box in the S. aurata PFKFB1 promoter was confirmed by chromatin immunoprecipitation assays. There is a great deal of evidence for a postprandial rise of PFKB1 mRNA levels in fish and rats. Consistently, starved-to-fed transition and treatment with glucose or insulin increased SREBP-1 immunodetectable levels, SREBP-1 association to PFKFB1 promoter, and PFKFB1 mRNA levels in the piscine liver. Our findings demonstrate involvement of SREBP-1a in the transcriptional activation of PFKFB1, and we conclude that SREBP-1a may exert a key role mediating postprandial activation of PFKFB1 transcription.

Assessing intellectual capital in a research institute environment in steel industry

Diplomityön tavoitteena oli tutkia aineettoman pääoman hallintaa terästeollisuuden tutkimuslaitosympäristössä, sekä luoda mittaristo kuvaamaan sen tilaa ja kehitystä. Työn rajausten mukaan aineeton pääoma jaettiin Annie Brookingin jaottelun mukaan, ja tarkasteluun valittiin teollisoikeudet sekä henkilöstövoimavarat. Empiirinen osa työstä perustui pääasiassa haastatteluihin sekä Case –yrityksen sisäisiin julkaisuihin. Näitä pyrittiin tulkitsemaan kvalitatiivisten tutkimusmenetelmien oppien mukaan. Tutkimustulosten perusteella voidaan todeta teollisoikeuksien hyödyntämisen olevan suhteellisen vähäistä terästeollisuuden parissa. Syitä tähän on työssä käsitelty melko laajasti. Henkilöstön kehittäminen pitkällä tähtäimellä sekä tiedonkulun tehostaminen henkilöstön keskuudessa ovat puolestaan asioita, joihin kiinnitetään kyseisellä teollisuudenalalla kiitettävässä määrin huomiota. Työn loppupuolella esitelty aineettoman pääoman mittaristo tarjoaa yritykselle työkalun, jonka avulla voidaan monipuolisesti arvioida tätä yrityksille nykyisin elintärkeää asiaa. Lopullista ratkaisumallia se ei aineettoman pääoman mittaamiseen tarjoa, mutta luo kuitenkin tärkeän ensiaskeleen kohti tulevaisuuden yhä kehittyneempiä mittausmenetelmiä.

The contribution of real-time mirror reflections of motor actions on virtual body ownership in an immersive virtual environment

This paper reports an experiment that investigated people"s body ownership of an avatar that was observed in a virtual mirror. Twenty subjects were recruited in a within-groups study where 10 first experienced a virtual character that synchronously reflected their upper-body movements as seen in a virtual mirror, and then an asynchronous condition where the mirror avatar displayed prerecorded actions, unrelated to those of the participant. The other 10 subjects experienced the conditions in the opposite order. In both conditions the participant could carry out actions that led to elevation above ground level, as seen from their first person perspective and correspondingly in the mirror. A rotating virtual fan eventually descended to 2m above the ground. The hypothesis was that synchronous mirror reflection would result in higher subjective sense of ownership. A questionnaire analysis showed that the body ownership illusion was significantly greater for thesynchronous than asynchronous condition. Additionally participants in the synchronous condition avoided collision with the descending fan significantly more often than those in the asynchronous condition. The results of this experiment are put into context within similar experiments on multisensory correlation and body ownership within cognitive neuroscience.

Bystander responses to a violent incident in an immersive virtual environment

Under what conditions will a bystander intervene to try to stop a violent attack by one person on another? It is generally believed that the greater the size of the crowd of bystanders, the less the chance that any of them will intervene. A complementary model is that social identity is critical as an explanatory variable. For example, when the bystander shares common social identity with the victim the probability of intervention is enhanced, other things being equal. However, it is generally not possible to study such hypotheses experimentally for practical and ethical reasons. Here we show that an experiment that depicts a violent incident at life-size in immersive virtual reality lends support to the social identity explanation. 40 male supporters of Arsenal Football Club in England were recruited for a two-factor between-groups experiment: the victim was either an Arsenal supporter or not (in-group/out-group), and looked towards the participant for help or not during the confrontation. The response variables were the numbers of verbal and physical interventions by the participant during the violent argument. The number of physical interventions had a significantly greater mean in the ingroup condition compared to the out-group. The more that participants perceived that the Victim was looking to them for help the greater the number of interventions in the in-group but not in the out-group. These results are supported by standard statistical analysis of variance, with more detailed findings obtained by a symbolic regression procedure based on genetic programming. Verbal interventions made during their experience, and analysis of post-experiment interview data suggest that in-group members were more prone to confrontational intervention compared to the out-group who were more prone to make statements to try to diffuse the situation.

Cost Estimation of Engineering Environment Services

Työn tavoitteena oli kehittää tutkittavan insinööriyksikön projektien kustannusestimointiprosessia, siten että yksikön johdolla olisi tulevaisuudessa käytettävänään tarkempaa kustannustietoa. Jotta tämä olisi mahdollista, ensin täytyi selvittää yksikön toimintatavat, projektien kustannusrakenteet sekä kustannusatribuutit. Tämän teki mahdolliseksi projektien kustannushistoriatiedon tutkiminen sekä asiantuntijoiden haastattelu. Työn tuloksena syntyi kohdeyksikön muiden prosessien kanssa yhteensopiva kustannusestimointiprosessi sekä –malli.Kustannusestimointimenetelmän ja –mallin perustana on kustannusatribuutit, jotka määritellään erikseen tutkittavassa ympäristössä. Kustannusatribuutit löydetään historiatietoa tutkimalla, eli analysoimalla jo päättyneitä projekteja, projektien kustannusrakenteita sekä tekijöitä, jotka ovat vaikuttaneet kustannusten syntyyn. Tämän jälkeen kustannusatribuuteille täytyy määritellä painoarvot sekä painoarvojen vaihteluvälit. Estimointimallin tarkuutta voidaan parantaa mallin kalibroinnilla. Olen käyttänyt Goal – Question – Metric (GQM) –menetelmää tutkimuksen kehyksenä.

Integrating Socket Server to the Workstation Environment of Series 40 Platform

Shortening development times of mobile phones are also accelerating the development times of mobile phone software. New features and software components should be partially implemented and tested before the actual hardware is ready. This brings challenges to software development and testing environments, especially on the user interface side. New features should be able to be tested in an environment, which has a look and feel like a real phone. Simulation environments are used to model real mobile phones. This makes possible to execute software in a mobile phone that does not yet exist. The purpose of this thesis is to integrate Socket Server software component to Series 40 simulation environments on Linux and Windows platforms. Socket Server provides TCP/IP connectivity for applications. All other software and hardware components below Socket Server do not exist in simulation environments. The scope of this work is to clarify how that can be done without connectivity problems, including design, implementation and testing phases.

Development of the Simulation Environment for the Mobile Phone User Interfaces

Diplomityössä käsitellään Nokia Mobile Phonesin matkapuhelimien käyttöliittymäohjelmistojen suunnittelu-ja testausympäristön kehitystä. Ympäristöön lisättiin kaksi ohjelmistomodulia avustamaan simulointia ja versionhallintaa. Visualisointityökalulla matkapuhelimen toiminta voidaan jäljittää suunnittelu- kaavioihin tilasiirtyminä, kun taas vertailusovelluksella kaavioiden väliset erot nähdään graafisesti. Kehitetyt sovellukset parantavat käyttöliittymien suunnitteluprosessia tehostaen virheiden etsintää, optimointia ja versionhallintaa. Visualisointityökalun edut ovat merkittävät, koska käyttöliittymäsovellusten toiminta on havaittavissa suunnittelu- kaavioista reaaliaikaisen simuloinnin yhteydessä. Näin virheet ovat välittömästi paikannettavissa. Lisäksi työkalua voidaan hyödyntää kaavioita optimoitaessa, jolloin sovellusten kokoja muistintarve pienenee. Graafinen vertailutyökalu tuo edun rinnakkaiseen ohjelmistosuunnitteluun. Eri versioisten suunnittelukaavioiden erot ovat nähtävissä suoraan kaaviosta manuaalisen vertailun sijaan. Molemmat työkalut otettiin onnistuneesti käyttöön NMP:llä vuoden 2001 alussa.

Novel therapeutic strategies targeting regulatory T cells and downstream TCR signaling in transplantation

Avec plus de 100000 transplantations d'organes solides (TOS) par année dans le monde, la transplantation d'organes reste actuellement l'un des meilleurs traitements disponibles pour de nombreuses maladies en phase terminale. Bien que les médicaments immunosuppresseurs couramment utilisés soient efficaces dans le contrôle de la réponse immune engendrant le rejet aigu d'une greffe, la survie du greffon à long terme ainsi que la présence d'effets secondaires indésirables restent un enjeu considérable en clinique. C'est pourquoi il est nécessaire de trouver de nouvelles approches thérapeutiques innovantes permettant de contrôler la réponse immunitaire et ainsi d'améliorer les résultats à long terme. L'utilisation des lymphocytes T régulateurs (Treg), suppresseurs naturels de la réponse inflammatoire, a fait l'objet de nombreuses études ces dix dernières années, et pourrait être considérée comme un moyen intéressant d'améliorer la tolérance immunologique de la greffe. Cependant, l'un des obstacles de l'utilisation des Treg comme agent thérapeutique est leur nombre insuffisant non seulement en conditions normales, mais en particulier lors d'une forte réponse immune avec expansion de cellules immunitaires alloréactives. En raison des limitations techniques connues pour l'induction des Treg ex-vivo ou in vitro, nous avons dédié la première partie du travail de thèse à la détermination de l'efficacité de l'induction des Treg in vivo grâce à l'utilisation d'un complexe protéique IL-2/JES6-1 (IL2c). Nous avons montré que l'expansion des Treg par IL2c permettait d'augmenter la survie du greffon sur un modèle murin de transplantation de peau avec mismatch entre le donneur et le receveur pour le complexe majeur d'histocompatibilité (CMH). De plus, nous avons vu qu'en combinant IL2c à une inhibition à court terme de la voie de co-stimulation CD40L-CD40 (anti-CD154/MRl, administré au moment de la transplantation) pour empêcher l'activation des lymphocytes T, il est possible d'induire une tolérance robuste à long terme. Finalement, nos résultats soulignent l'importance de cibler une voie de co-stimulation bien particulière. En effet, l'utilisation d'IL2c combinée au blocage de la co-stimulation CD28-B7.1/2 (CTLA-4 Ig) n'induit qu'une faible prolongation de la survie de la greffe et n'induit pas de tolérance. L'application chez l'humain des traitements induisant la tolérance dans des modèles expérimentaux murins ou de primates n'a malheureusement pas montré de résultats probants en recherche clinique ; une des principales raisons étant la présence de lymphocytes B et T mémoires provenant du systeme d immunité acquise. C est pourquoi nous avons testé si la combinaison d'IL2c et MR1 améliorait la survie de la greffe dans des souris pré¬sensibilisées. Nous avons trouvé qu'en présence de lymphocytes B et T mémoires alloréactifs, l'utilisation d'IL2c et MR1 permettait une amélioration de la survie de la greffe de peau des souris immunocompétentes mais comparé aux souris receveuses naïves, aucune tolérance n'a pu être induite. Toutefois, l'ajout d'un traitement anti-LFA-1 (permettant de bloquer la circulation des lymphocytes T activées) a permis d'améliorer de manière significative la survie de la greffe. Cependant, le rejet chronique, dû à la présence de lymphocytes B activés/mémoires et la production d'anticorps donneur-spécifiques, n'a pas pu être évité. Cibler l'activation des lymphocytes T est la stratégie immunothérapeutique prépondérente après une TOS. C'est pourquoi dans la deuxième partie de cette thèse nous nous sommes intéressés au système de signalisation d'un récepteur des lymphocytes T qui dépend de la paracaspase Malti en tant que nouvelle stratégie immunosuppressive pour le contrôle des lymphocytes T alloréactifs. Nous avons montré que bien que l'inhibition de la signalisation du lymphocyte T en aval de Malti induise une tolérance envers un greffon de peau avec incompatibilités antigéniques mineures, cela ne permet cependant qu'une régulation partielle de l'alloréponse contre des antigènes du CMH. Nous nous sommes aussi intéressés spécifiquement à l'activité protéolytique de Malti. L'inhibition constitutive de l'activité protéolytique de Malti chez les souris Malti-ki s'est révélée délétère pour l'induction de la tolérance car elle diminue la fonction des Treg et augmente l'alloréactivité des cellules Thl. Cependant, lors de l'utilisation d'un inhibiteur peptidique de l'activité protéase de Malti in vitro, il a été possible d'observer une atténuation de l'alloéactivité des lymphocytes T ainsi qu'un maintien de la population des Treg existants. Ces résultats nous laissent penser que des études plus poussées sur le rôle de la signalisation médiée par Malti seraient à envisager dans le domaine de la transplantation. En résumé, les résultats obtenus durant cette thèse nous ont permis d'élucider certains mécanismes immunologiques propres à de nouvelles stratégies thérapeutiques potentielles dont le but est d'induire une tolérance lors de TOS. De plus, ces résultats nous ont permis de souligner l'importance d'utiliser des modèles davantage physiologiques contenant, notamment en tenant compte des lymphocytes B et T mémoires alloréactifs. -- Organ transplantation remains the best available treatment for many forms of end-stage organ diseases, with over 100,000 solid organ transplantations (SOT) occurring worldwide eveiy year. Although the available immunosuppressive (IS) drugs are efficient in controlling acute immune activation and graft rejection, the off-target side effects as well as long-term graft and patient survival remain a challenge in the clinic. Hence, innovative therapeutic approaches are needed to improve long-term outcome across immunological barriers. Based on extensive experimental data obtained over the last decade, it is tempting to consider immunotherapy using Treg; the natural suppressors of overt inflammatory responses, in promoting transplantation tolerance. The first hurdle for the therapeutic use of Treg is their insufficient numbers in non- manipulated individuals, in particular when facing strong immune activation and expanding alloreactive effector cells. Because of the limitations associated with current protocols aiming at ex-vivo expansion or in vitro induction of Treg, the aim of the first part of this thesis was to determine the efficacy of direct in vivo expansion of Treg using the IL-2/JES6- 1 immune complex (IL2c). We found that whilst IL2c mediated Treg expansion alone allowed the prolonged graft survival of fìlli MHC-mismatched skin grafts, its combination with short-term CD40L-CD40 co-stimulation blockade (anti-CD 154/MR1) to inhibit T cell activation administered at the time of transplantation was able to achieve long-term robust tolerance. This study also highlighted the importance of combining Treg based therapies with the appropriate co-stimulation blockade as a combination of IL2c and CD28-B7.1/2 co- stimulation blockade (CTLA-4 Ig) only resulted in slight prolongation of graft survival but not tolerance. The translation of tolerance induction therapies modelled in rodents into non-human primates or into clinical trials has seldom been successful. One main reason being the presence of pre-existing memory T- and B-cells due to acquired immunity in humans versus laboratory animals. Hence, we tested whether IL2c+MRl could promote graft survival in pre-sensitized mice. We found that in the presence of alloreactive memory T- and B-cells, IL2c+MRl combination therapy could prolong MHC-mismatched skin graft survival in immunocompetent mice but tolerance was lost compared to the naïve recipients. The addition of anti-LF A-1 treatment, which prevents the trafficking of memory T cells worked synergistically to significantly further enhance graft survival. However, late rejection mediated by activated/memory B cells and persistent donor-specific alloantibodies still occurred. Immunotherapeutic strategies targeting the activation of T cells are the cornerstone in the current immunosuppressive management after SOT. Therefore, in the next part of this thesis we investigated the paracaspase Malti-dependent T-cell receptor signalling as a novel immunosuppressive strategy to control alloreactive T cells in transplantation. We observed that although the inhibition of Malti downstream T signalling lead to tolerance of a minor H- mismatch skin grafts, it was however not sufficient to regulate alloresponses against MHC mismatches and only prolonged graft survival. Furthermore, we investigated the potential of more selectively targeting the protease activity of Malti. Constitutive inhibition of Malti protease activity in Malti-ki mice was detrimental to tolerance induction as it diminished Treg function and increased Thl alloreactivity. However, when using a small peptide inhibitor of Malti proteolytic activity in vitro, we observed an attenuation of alloreactive T cells and sparing of the pre-existing Treg pool. This indicates that further investigation of the role of Malti signalling in the field of transplantation is required. Collectively, the findings of this thesis provide immunological mechanisms underlying novel therapeutic strategies for the promotion of tolerance in SOT. Moreover, we highlight the importance of testing tolerance induction therapies in more physiological models with pre-existing alloreactive memory T and B cells.