Between Immunology And Tolerance: Controlling Immune Responses Employing Tolerogenic Dendritic Cells

Dendritic cells (DCs) are the most efficient antigen presenting cells, they provide co-stimulation, are able to secrete various proinflammatory cytokines and therefore play a pivotal role in shaping adaptive immune responses. Moreover, they are important for the promotion and maintenance of central and peripheral tolerance through several mechanisms like the induction of anergy or apoptosis in effector T cells or by promoting regulatory T cells. The murine CD8α+ (MuTu) dendritic cell line was previously derived and described in our laboratory. The MuTu cell line has been shown to maintain phenotypical and functional characteristics of endogenous CD8α+ DCs. They are able to cross-present exogenous antigens to CD8+ T cells and produce interleukin (IL-) 12 upon engagement of Toll like receptors. The cell line constitutes an infinite source of homogenous, phenotypically well-defined dendritic cells. This allows us to investigate the role and potential of specific molecules in the induction as well as regulation of immune responses by DCs in a rational and standardized way. In a first project the MuTu dendritic cell line was transduced in order to stably express the immunosuppressive molecules IL-10, IL-35 or the active form of TGF-β (termed IL-10+DC, IL-35+DC or actTGFβ+DC). We investigated the capability of these potentially suppressive or tolerogenic dendritic cell lines to induce immune tolerance and explore the mechanisms behind tolerance induction. The expression of TGF-β by the DC line did not affect the phenotype of the DCs itself. In contrast, IL-10+ and IL-35+DCs were found to exhibit lower expression of co-stimulatory molecules and MHC class I and II, as well as reduced secretion of pro-inflammatory cytokines upon activation. In vitro co-culture with IL-35+, IL10+ or active TGFβ+ DCs interfered with function and proliferation of CD4+ and CD8+ T cells. Furthermore, IL-35 and active TGF-β expressing DC lines induced regulatory phenotype on CD4+ T cells in vitro without or with expression of Foxp3, respectively. In different murine cancer models, vaccination with IL-35 or active TGF-β expressing DCs resulted in faster tumor growth. Interestingly, accelerated tumor growth could be observed when IL-35-expressing DCs were injected into T cell-deficient RAG-/- mice. IL-10expressing DCs however, were found to rather delay tumor growth. Besides the mentioned autocrine effects of IL-35 expression on the DC line itself, we surprisingly observed that the expression of IL-35 or the addition of IL-35 containing medium enhances neutrophil survival and induces proliferation of endothelial cells. Our findings indicate that the cytokine IL-35 might not only be a potent regulator of adaptive immune responses, but it also implies IL-35 to mediate diverse effects on an array of cellular targets. This abilities make IL-35 a promising target molecule not only for the treatment of auto-inflammatory disease but also to improve anti-cancer immunotherapies. Indeed, by applying active TGFβ+ in murine autoimmune encephalitis we were able to completely inhibit the development of the disease, whereas IL-35+DCs reduced disease incidence and severity. Furthermore, the preventive transfer of IL-35+DCs delayed rejection of transplanted skin to the same extend as the combination of IL-10/actTGF-β expressing DCs. Thus, the expression of a single tolerogenic molecule can be sufficient to interfere with the adequate activation and function of dendritic cells and of co-cultured T lymphocytes. The respective mechanisms of tolerance induction seem to be different for each of the investigated molecule. The application of a combination of multiple tolerogenic molecules might therefore evoke synergistic effects in order to overcome (auto-) immunity. In a second project we tried to improve the immunogenicity of dendritic cell-based cancer vaccines using two different approaches. First, the C57BL/6 derived MuTu dendritic cell line was genetically modified in order to express the MHC class I molecule H-2Kd. We hypothesized that the expression of BALB/c specific MHC class I haplotype (H-2Kd) should allow the priming of tumor-specific CD8+ T cells by the otherwise allogeneic dendritic cells. At the same time, the transfer of these H-2Kd+ DCs into BALB/c mice was thought to evoke a strong inflammatory environment that might act as an "adjuvant", helping to overcome tumor induced immune suppression. Using this so called "semi-allogeneic" vaccination approach, we could demonstrate that the delivery of tumor lysate pulsed H-2Kd+ DCs significantly delayed tumor growth when compared to autologous or allogeneic vaccination. However, we were not able to coherently elucidate the cellular mechanisms underlying the observed effect. Second, we generated MuTu DC lines which stably express the pro-inflammatory cytokines IL-2, IL-12 or IL-15. We investigated whether the combination of DC vaccination and local delivery of pro-inflammatory cytokines might enhance tumor specific T cell responses. Indeed, we observed an enhanced T cell proliferation and activation when they were cocultured in vitro with IL-12 or IL-2-expressing DCs. But unfortunately we could not observe a beneficial or even synergistic impact on tumor development when cytokine delivery was combined with semi-allogeneic DC vaccination.

What interventions facilitate client progress through the assimilation model? A task analysis of interventions in the psychodynamic treatment of depression.

A variation of task analysis was used to build an empirical model of how therapists may facilitate client assimilation process, described in the Assimilation of Problematic Experiences Scale. A rational model was specified and considered in light of an analysis of therapist in-session performances (N = 117) drawn from six inpatient therapies for depression. The therapist interventions were measured by the Comprehensive Psychotherapeutic Interventions Rating Scale. Consistent with the rational model, confronting interventions were particularly useful in helping clients elaborate insight. However, rather than there being a small number of progress-related interventions at lower levels of assimilation, therapists' use of interventions was broader than hypothesized and drew from a wide range of therapeutic approaches. Concerning the higher levels of assimilation, there was insufficient data to allow an analysis of the therapist's progress-related interventions.

Using vegetation to characterize the avalanche to Canal del Roc Roig, Vall de Núria (Eastern Pyrenees, Spain)

Avalanche hazard maps of high accuracy are difficult to produce. For land-use planning and management purposes, a good knowledge of extreme run-out zones and frequencies of avalanches is required. In the present work, vegetation recognition (especially focused on Pinus uncinata trees) and dendrochronological techniques are used to characterize avalanches that have occurred in historical times, helping to determine both the extent of large or extreme avalanches and their occurrence in time. Vegetation was studied at the Canal del Roc Roig (eastern Pyrenees, Spain) avalanche path. The avalanches descending this path affect the railway that reaches the Vall de Núria resort and the run-up to the opposite slope. During winter 1996, two important avalanches affecting this path were well documented. These are compared with the results of the vegetation study, consisting of an inventory of flora, the recording of vegetation damages along eight transverse profiles at different altitudes on the path and a dendrochronological sampling campaign. The data obtained contributed to a characterization of the predominant snow accumulation in the starting zone, the 1996 avalanches and the range of frequencies of large avalanches. Also, traces of avalanches that increase the path mapped in the avalanche paths map published by the Institut Cartogràfic de Catalunya in 2000 were identified, improving the initial existing information.

Using vegetation to characterize the avalanche to Canal del Roc Roig, Vall de Núria (Eastern Pyrenees, Spain)

Avalanche hazard maps of high accuracy are difficult to produce. For land-use planning and management purposes, a good knowledge of extreme run-out zones and frequencies of avalanches is required. In the present work, vegetation recognition (especially focused on Pinus uncinata trees) and dendrochronological techniques are used to characterize avalanches that have occurred in historical times, helping to determine both the extent of large or extreme avalanches and their occurrence in time. Vegetation was studied at the Canal del Roc Roig (eastern Pyrenees, Spain) avalanche path. The avalanches descending this path affect the railway that reaches the Vall de Núria resort and the run-up to the opposite slope. During winter 1996, two important avalanches affecting this path were well documented. These are compared with the results of the vegetation study, consisting of an inventory of flora, the recording of vegetation damages along eight transverse profiles at different altitudes on the path and a dendrochronological sampling campaign. The data obtained contributed to a characterization of the predominant snow accumulation in the starting zone, the 1996 avalanches and the range of frequencies of large avalanches. Also, traces of avalanches that increase the path mapped in the avalanche paths map published by the Institut Cartogràfic de Catalunya in 2000 were identified, improving the initial existing information.

Automatic target validation based on neuroscientific literature mining for tractography.

Target identification for tractography studies requires solid anatomical knowledge validated by an extensive literature review across species for each seed structure to be studied. Manual literature review to identify targets for a given seed region is tedious and potentially subjective. Therefore, complementary approaches would be useful. We propose to use text-mining models to automatically suggest potential targets from the neuroscientific literature, full-text articles and abstracts, so that they can be used for anatomical connection studies and more specifically for tractography. We applied text-mining models to three structures: two well-studied structures, since validated deep brain stimulation targets, the internal globus pallidus and the subthalamic nucleus and, the nucleus accumbens, an exploratory target for treating psychiatric disorders. We performed a systematic review of the literature to document the projections of the three selected structures and compared it with the targets proposed by text-mining models, both in rat and primate (including human). We ran probabilistic tractography on the nucleus accumbens and compared the output with the results of the text-mining models and literature review. Overall, text-mining the literature could find three times as many targets as two man-weeks of curation could. The overall efficiency of the text-mining against literature review in our study was 98% recall (at 36% precision), meaning that over all the targets for the three selected seeds, only one target has been missed by text-mining. We demonstrate that connectivity for a structure of interest can be extracted from a very large amount of publications and abstracts. We believe this tool will be useful in helping the neuroscience community to facilitate connectivity studies of particular brain regions. The text mining tools used for the study are part of the HBP Neuroinformatics Platform, publicly available at http://connectivity-brainer.rhcloud.com/.

Konttipaikannuksen toteutus konttitrukkiin

Diplomityössä perehdytään konttipaikannustekniikkaan, historiaan ja useisiin tekniikoihin joilla järjestelmiä voidaan toteuttaa. Huomiota on kiinnitetty erityisesti GPS paikannukseen ja satelliittien hyödyntämiseen merkintälaskussa. Lisäksi huomiota on kiinnitetty eri anturi tyyppeihin joita tullaan tarvitsemaan. Käytännön sovelluksena työssä on toteutettu konttipaikannus järjestelmä konttitrukkiin. Työssä on tarkasteltu eri teknisiä mahdollisuuksia ja niiden yhdistämistä niin, että saadaan toimiva sovellus.

Robots and robotic systems for the electronics and telecommunication industry

Tämä diplomityö tutkii elektroniikka- ja telekommunikaatioteollisuutta sekä siihen läheisesti liittyviä robotteja ja robottijärjestelmiä. Tavoitteena on määrittää E&T-teollisuuden prosesseihin soveltuvien robottien testausmenetelmä. Tavoitteena on myös selvittää kahden ABB:n robotin soveltuvuutta E&T-teollisuuden tarpeisiin. Muutamia systemaattisia valmistusjärjestelmien suunnitteluun soveltuvia menetelmiä ja apuvälineitä on myös käsitelty. Alussa työ keskittyy elektroniikka- ja telekommunikaatioteollisuuden nykytilan tutkimiseen sekä siellä vallitsevien ja ennustettujen trendien kartoitukseen. Kohdat “Collaborative manufacturing” ja E&T-teollisuuden valmistusjärjestelmille asettamat vaatimukset käydään yksityiskohtaisesti läpi. Tutkimuksen pääkohteina ovat robotit, erityisesti ABB:n IRB 140 ja IRB 340 sekä robottien testausmenetelmän määrittäminen. Työssä käydään läpi IRB 340:llä suoritetut testit, jotka tehtiin sekä konenäköjärjestelmää apuna käyttäen että ilman. Myös TTKK:lla suoritetut robottitestit on käyty läpi. Robottien testituloksia on analysoitu ja vertailtu muihin robotteihin. Testausmenetelmät perustuvat ISO 9283 standardiin. Viimeinen osa työstä esittelee robottijärjestelmien systemaattiseen suunnitteluun soveltuvia menetelmiä ja apuvälineitä. Esillä ovat mm. Modular function deployment (MFD) ja The system design method (SDM).

An Empirical Investigation of Defense Interpretation Depth, Defensive Functioning, and Alliance Strength in Psychodynamic Psychotherapy.

The present study examined the relationship between depth of defense interpretations by therapists, and patient defensive functioning, on the therapeutic alliance in a sample of 36 patients undergoing short-term dynamic psychotherapy. Defense interpretation depth was defined as the degree to which therapist interpretations contained information regarding the motivation for patient defenses and historical origins of the defensive processes (Greensen, 1967). Mean depth of interpretation was compared between sessions that were identified beforehand as either high-alliance or low-alliance sessions using the Helping Alliance Questionnaire (HAq-II: Luborsky et al., 1996). Results indicated that defensive functioning was correlated to defense interpretation depth in low-alliance sessions. Moreover, mean depth of interpretation was also higher in low-alliance sessions, pointing to the possible "destabilizing" effects that these interpretations may have on both defensive functioning and the therapeutic alliance. These results are discussed within the context of previous studies of therapeutic technique in dynamic psychotherapy.

Tilitoimisto pienyrityksen taloushallinnon asiantuntijana

Tämän tutkielman päätavoitteena oli selvittää, miten tilitoimistojen tuottamat palvelut vastaavat pienyrityksen johtamisen, ja erityisesti talousjohtamisen tarpeita ja auttavat yritystä menestymään. Alatavoitteina oli selvittää ja analysoida pienyrityksen johtamista, taloushallintoa ja menestystekijöitä, sekä asiantuntijapalveluita ja tilitoimistoja ja niiden palvelutarjonnan rakennetta. Tutkimusmenetelmänä käytettiin käsiteanalyyttistä tutkimusotetta. Aikaisemmin julkaistuun tutkimusdoktriiniin, joka sisälsi niin teoreettisia kuin empiirisiäkin tieteellisiä tutkimuksia, ammattikirjallisuutta ja -lehtiartikkeleita, perehtymällä pyrittiin ratkaisemaan asetetut tavoitteet. Tutkimustulos osoittaa, että tilitoimistoilla on hyvät valmiudet palvella asiantuntevasti pienyritystä ja auttaa sitä menestymään tuottamalla tarvittavaa taloushallinnon informaatiota ja muita palveluita. Yrityksen rahoituksen laskentatoimen osalta tilitoimistojen palvelut vastaavat hyvin pienyrityksen johtamisessa tarvittavaa tietoa, johdon laskentatoimen osalta tuotettavat palvelut ovat vielä suurelta osin alikehittyneitä johtuen osaksi tilitoimistojen osaamistasosta, mutta myös pienyritysten haluttomuudesta vastaanottaa laskentatoimen tuottamaa informaatiota ja johdon heikosta talous- asioiden hallinnasta.

Evolutionary dynamics of collective action in spatially structured populations.

Many models proposed to study the evolution of collective action rely on a formalism that represents social interactions as n-player games between individuals adopting discrete actions such as cooperate and defect. Despite the importance of spatial structure in biological collective action, the analysis of n-player games games in spatially structured populations has so far proved elusive. We address this problem by considering mixed strategies and by integrating discrete-action n-player games into the direct fitness approach of social evolution theory. This allows to conveniently identify convergence stable strategies and to capture the effect of population structure by a single structure coefficient, namely, the pairwise (scaled) relatedness among interacting individuals. As an application, we use our mathematical framework to investigate collective action problems associated with the provision of three different kinds of collective goods, paradigmatic of a vast array of helping traits in nature: "public goods" (both providers and shirkers can use the good, e.g., alarm calls), "club goods" (only providers can use the good, e.g., participation in collective hunting), and "charity goods" (only shirkers can use the good, e.g., altruistic sacrifice). We show that relatedness promotes the evolution of collective action in different ways depending on the kind of collective good and its economies of scale. Our findings highlight the importance of explicitly accounting for relatedness, the kind of collective good, and the economies of scale in theoretical and empirical studies of the evolution of collective action.

Fusion weld metal solidification:Continuum from weld interface to centerline

We present a brief résumé of the history of solidification research and key factors affecting the solidification of fusion welds. There is a general agreement of the basic solidification theory, albeit differing - even confusing - nomenclatures do exist, and Cases 2 and 3 (the Chalmers' basic boundary conditions for solidification, categorized by Savage as Cases) are variably emphasized. Model Frame, a tool helping to model the continuum of fusion weld solidification from start to end, is proposed. It incorporates the general solidification models, of which the pertinent ones are selected for the actual modeling. The basic models are the main solidification Cases 1…4. These discrete Cases are joined with Sub-Cases: models of Pfann, Flemings and others, bringing needed Sub-Case variables into the model. Model Frame depicts a grain growing from the weld interface to its centerline. Besides modeling, the Model Frame supports education and academic debate. The new mathematical modeling techniques will extend its use into multi-dimensional modeling, introducing new variables and increasing the modeling accuracy. We propose a model: melting/solidification-model (M/S-model) - predicting the solute profile at the start of the solidification of a fusion weld. This Case 3-based Sub-Case takes into account the melting stage, the solute back-diffusion in the solid, and the growth rate acceleration typical to fusion welds. We propose - based on works of Rutter & Chalmers, David & Vitek and our experimental results on copper - that NEGS-EGS-transition is not associated only with cellular-dendritic-transition. Solidification is studied experimentally on pure and doped copper with welding speed range from 0 to 200 cm/min, with one test at 3000 cm/min. Found were only planar and cellular structures, no dendrites - columnar or equiaxed. Cell sub structures: rows of cubic elements we call "cubelettes", "cell-bands" and "micro-cells", as well as an anomalous crack morphology "crack-eye", were detected, as well as microscopic hot crack nucleus we call "grain-lag cracks", caused by a grain slightly lagging behind its neighbors in arrival to the weld centerline. Varestraint test and R-test revealed a change of crack morphologies from centerline cracks to grainand cell boundary cracks with an increasing welding speed. High speed made the cracks invisible to bare eye and hardly detectable with light microscope, while electron microscope often revealed networks of fine micro-cracks.

Non-invasive prenatal testing for aneuploidy and beyond: challenges of responsible innovation in prenatal screening.

This paper contains a joint ESHG/ASHG position document with recommendations regarding responsible innovation in prenatal screening with non-invasive prenatal testing (NIPT). By virtue of its greater accuracy and safety with respect to prenatal screening for common autosomal aneuploidies, NIPT has the potential of helping the practice better achieve its aim of facilitating autonomous reproductive choices, provided that balanced pretest information and non-directive counseling are available as part of the screening offer. Depending on the health-care setting, different scenarios for NIPT-based screening for common autosomal aneuploidies are possible. The trade-offs involved in these scenarios should be assessed in light of the aim of screening, the balance of benefits and burdens for pregnant women and their partners and considerations of cost-effectiveness and justice. With improving screening technologies and decreasing costs of sequencing and analysis, it will become possible in the near future to significantly expand the scope of prenatal screening beyond common autosomal aneuploidies. Commercial providers have already begun expanding their tests to include sex-chromosomal abnormalities and microdeletions. However, multiple false positives may undermine the main achievement of NIPT in the context of prenatal screening: the significant reduction of the invasive testing rate. This document argues for a cautious expansion of the scope of prenatal screening to serious congenital and childhood disorders, only following sound validation studies and a comprehensive evaluation of all relevant aspects. A further core message of this document is that in countries where prenatal screening is offered as a public health programme, governments and public health authorities should adopt an active role to ensure the responsible innovation of prenatal screening on the basis of ethical principles. Crucial elements are the quality of the screening process as a whole (including non-laboratory aspects such as information and counseling), education of professionals, systematic evaluation of all aspects of prenatal screening, development of better evaluation tools in the light of the aim of the practice, accountability to all stakeholders including children born from screened pregnancies and persons living with the conditions targeted in prenatal screening and promotion of equity of access.

The Role of the Subjectivist Position in the Probabilization of Forensic Science

This paper is concerned with the contribution of forensic science to the legal process by helping reduce uncertainty. Although it is now widely accepted that uncertainty should be handled by probability because it is a safeguard against incoherent proceedings, there remain diverging and conflicting views on how probability ought to be interpreted. This is exemplified by the proposals in scientific literature that call for procedures of probability computation that are referred to as "objective," suggesting that scientists ought to use them in their reporting to recipients of expert information. I find such proposals objectionable. They need to be viewed cautiously, essentially because ensuing probabilistic statements can be perceived as making forensic science prescriptive. A motivating example from the context of forensic DNA analysis will be chosen to illustrate this. As a main point, it shall be argued that such constraining suggestions can be avoided by interpreting probability as a measure of personal belief, that is, subjective probability. Invoking references to foundational literature from mathematical statistics and philosophy of science, the discussion will explore the consequences of this interdisciplinary viewpoint for the practice of forensic expert reporting. It will be emphasized that-as an operational interpretation of probability-the subjectivist perspective enables forensic science to add value to the legal process, in particular by avoiding inferential impasses to which other interpretations of probability may lead. Moreover, understanding probability from a subjective perspective can encourage participants in the legal process to take on more responsibility in matters regarding the coherent handling of uncertainty. This would assure more balanced interactions at the interface between science and the law. This, in turn, provides support for ongoing developments that can be called the "probabilization" of forensic science.

Stochasticity in economic losses increases the value of reputation in indirect reciprocity.

Recent theory predicts harsh and stochastic conditions to generally promote the evolution of cooperation. Here, we test experimentally whether stochasticity in economic losses also affects the value of reputation in indirect reciprocity, a type of cooperation that is very typical for humans. We used a repeated helping game with observers. One subject (the "Unlucky") lost some money, another one (the "Passer-by") could reduce this loss by accepting a cost to herself, thereby building up a reputation that could be used by others in later interactions. The losses were either stable or stochastic, but the average loss over time and the average efficiency gains of helping were kept constant in both treatments. We found that players with a reputation of being generous were generally more likely to receive help by others, such that investing into a good reputation generated long-term benefits that compensated for the immediate costs of helping. Helping frequencies were similar in both treatments, but players with a reputation to be selfish lost more resources under stochastic conditions. Hence, returns on investment were steeper when losses varied than when they did not. We conclude that this type of stochasticity increases the value of reputation in indirect reciprocity.

Palvelun laadun rooli brandistrategian kehittämisessä

Tutkimuksen päätavoitteena on selvittää palvelun laadun rooli brandistrategian kehittämisessä. Tutkimuksen tekeminen nähtiin tarpeelliseksi, sillä palvelun laadun roolia brandistrategian kehittämisessä ei ole aiemmin paljolti tutkittu. Normatiivisella case-tutkimuksella pyritään löytämään konkreettisia brandistrategian kehittämiskohteita case-yritykselle niin strategisella kuin myös operationaalisella tasolla. Case-yrityksen toimiala ja tilanne huomioiden normatiiviselle case-tutkimukselle luodaan perusta tutkimuksen teoreettisessa osassa sovellettavilla teoreettisilla näkemyksillä ja malleilla. Tutkimusta käsitellään sekä kvalitatiivisten että kvantitatiivisten tutkimusmenetelmien keinoin. Tutkimuksen empiirinen osio perustuu yrityksen henkilökunnan haastatteluihin, tutkijan havainnointiin sekä case—yrityksen asiakkaille suoritettavaan asiakastutkimukseen. Tutkimus osoittaa, että palvelun laadun rooli on merkittävässä asemassa palveluyritysten brandistrategian kehittämisessä ja on samalla tärkein yksittäinen asiakastyytyväisyyteen vaikuttava tekijä. Tutkimus tuo esille konkreettisia toimenpide-ehdotuksia, jotka auttavat case-yritystä kehittämään brandistrategiaansa ja auttaa siten yritystä saavuttamaan kilpailuetua muuttuvassa toimintaympäristössä.