Les lymphomes anaplasiques à grandes cellules (LAGC) sont des lymphomes non-hodgkiniens dits de type T ou nul, représentant la majorité des lymphomes T pédiatriques (20 à 30%). Dans plus de 80% des cas, une translocation chromosomique réciproque aboutissant à l'expression anormale de protéines chimères de type X-ALK qui arborent constitutivement et de manière anormale l'activité tyrosine-kinase ALK (Anaplastic Lymphoma Kinase) est le moteur de la tumorigenèse (LAGC dits "ALK+ "). L'une des particularités de ces lymphomes, mise en évidence par mon équipe, est le fait que B-Cell Lymphoma-2 (BCL-2) demeure indétectable dans les cas ALK+ contrairement aux cas ALK-. Ce point est d'autant plus surprenant que BCL-2, oncogène largement établi comme prototype de protéines anti-apoptotiques ainsi que régulateur clé de l'autophagie, est fortement surexprimé dans la majorité des lymphomes. A l'inverse, Human Antigen R (HuR) est surexprimée dans les LAGC (comme dans la plupart des cancers). Il a été démontré que cette protéine de liaison aux ARN participait au maintien du phénotype tumoral, et que sa localisation subcellulaire et ses fonctions dépendaient étroitement de son statut de phosphorylation, lequel est régulé par ALK dans les LAGC ALK+. Au niveau du cytoplasme, HuR permet de stabiliser et d'augmenter la traduction d'ARNm possédant, dans leur région 3'-UTR, des séquences riches en adénine et uridine (AU-rich elements, "ARE"). De manière plus générale, HuR a la capacité de dialoguer avec les microARNs (miARN), soit en empêchant leur action par compétition, soit à l'inverse en coopérant avec ces derniers et en promouvant ainsi leur fonction de régulateur négatif sur certains transcrits cibles communs. Le transcrit Bcl-2, dont l'expression est réprimée dans les LAGC ALK+, fait partie des cibles potentielles de HuR. Au cours de ma thèse, j'ai ainsi cherché à comprendre les mécanismes moléculaires mis en jeu dans la répression de l'expression de Bcl-2, en me focalisant sur le rôle de HuR et de miARN "partenaires" dans ce processus. Mes données semblent indiquer que ce mécanisme implique le recrutement par HuR du miR-34a sur l'ARNm Bcl-2, conduisant à la mise en silence de ce dernier. A l'inverse, quand l'activité tyrosine- kinase de ALK est inhibée, l'interaction entre HuR et le transcrit Bcl-2 diminue, ce qui limite le recrutement de miR-34a et conduit à une restauration de l'expression de cette oncogène majeur dans les cellules lymphomateuses. Dans le contexte des essais cliniques d'inhibiteurs ciblant l'activité tyrosine kinase de ALK tels que le Crizotinib, la question de cette ré-expression de BCL-2 éclaire d'une lumière nouvelle certains échecs thérapeutiques subis par cette molécule pourtant prometteuse. Je me suis donc également consacré, pendant ma thèse, à l'étude des conséquences de cette ré-expression de BCL-2 sur les LAGC ALK+ traités par le Crizotinib. Les résultats que j'ai obtenus in vitro et in vivo montrent que contrecarrer, par interférence à l'ARN, l'élévation du taux de BCL-2 consécutive au traitement par le Crizotinib, permet de potentialiser les effets de la drogue : cela se traduit en particulier par une potentialisation de la mort par apoptose induite par le traitement mais aussi, de manière fascinante, par une conversion de la réponse autophagique initialement cytoprotectrice et pro-tumorale en une autophagie incontrôlée et délétère, qui participe alors à l'effet thérapeutique accru de la drogue. De manière globale, ce travail permet d'envisager de nouvelles combinaisons et alternatives thérapeutiques pour les patients souffrants de LAGC ALK+, et illustre la complexité des régulations croisées entre processus apoptotiques et autophagiques. / Anaplastic large cell lymphoma (ALCL) are T/-null non-hodgkin lymphoma representing most of childhood T-cell lymphoma (up to 30%). More than 80% of cases bear reciprocal chromosomic translocation responsible for abnormal expression and constitutive activation of X-ALK type (Anaplastic Lymphoma Kinase) chimeric proteins (ALK+ ALCL). A striking characteristic of this lymphoma is that B-Cell Lymphoma-2 (BCL-2) remains undetectable in ALK+ cases compared to ALK- cases. This is all the more surprising as the BCL-2 oncogene, which is firmly established as a prototypic anti-apoptotic factor as well as a key autophagy regulator, has been shown to be overexpressed in a majority of lymphomas. On the other hand, the RNA-binding protein HuR (Human Antigen R) is overexpressed in ALCL (as in most cancers). It has been demonstrated that this protein was involved in the sustainability of the tumoral phenotype, and that its subcellular localization and functions were closely related to its phosphorylation status, which in turn heavily depends on ALK activity in ALK+ ALCL. In the cytoplasm, HuR has the ability to bind adenine and uridine-rich elements (ARE) located on the 3'-UTR of target mRNAs, and both protect them from degradation and increase their translation. From a general point of view, HuR is able to establish an interplay with microRNAs (miRNAs), either blocking them through competition, or actually cooperating with them and thus promote their function of negative regulators of gene expression on common target transcripts. The BCL-2 transcript, which expression seems to be silenced in ALK-expressing ALCL, has been described as a potential target of HuR. During my PhD work, I dedicated myself to understand the molecular mechanism at work in the silencing of BCL-2 expression with a focus on HuR and collaborating miRNA. The data I obtained point at a cooperation between HuR and miR-34a leading to the silencing of the BCL-2 transcript. However, when the ALK tyrosine kinase activity is inhibited, it appears the interaction between the BCL-2 mRNA diminishes, which limitates the miR-34a 's access to this transcript and ultimately results in a re-expression of the BCL-2 oncogene in these lymphoma cells. In the current context of clinical trials for ALK-targeting inhibitors, such as the Crizotinib, this BCL-2 re-expression observed upon ALK inhibition shed light on potential reasons behind some therapeutic failures that have recently been reported. Indeed, during my PhD work, I also studied the consequences of the BCL-2 re-expression observed in Crizotinib-treated cells. The data I obtained in vitro and in vivo show that, by blocking this re-expression using RNA interference, the Crizotinib anti-tumoral efficiency can be greatly potentiated. This potentiation took the form of an increase of apoptotic cell death induction and, interestingly, also affected the autophagic response triggered by the drug, making it switch from a cytoprotective- type, protumoral autophagic flux to an enhanced, deletary-type and tumor suppressive flux, adding to the therapeutic effect of the drug. This work in general provides insights for new therapeutic combinations that could potentially benefit to ALK+ ALCL patients, and illustrates the complex cross-regulations between apoptotic and autophagic pathway.

Les méthodes de contrôle de vibrations passives basées sur des absorbeurslinéaires ont été largement étudiées et ils ont aujourd'hui une vaste gammed'applications. Cependant, les absorbeurs linéaires n’étant efficaces quelorsqu'ils sont accordés à la fréquence que l'on veut contrôler, ils présententdes limites considérables quand ils sont appliqués à des systèmes possédantdes incertitudes sur les paramètres modaux ou ayant une fréquence propredépendante de la force extérieure. Dans cette thèse la réduction des vibrations dans les systèmes mécaniques à l'aide d'un absorbeur Nonlinear Energy Sink est étudiée. Le phénomène qui gouverne la physique de ce dispositif est appelé pompage énergétique (Targeted Energy Transfer) et il consiste en un transfert irréversible d'énergie du système principal vers le NES, où l’énergie est dissipée. Ce transfert d'énergie peut se produire pour une large gamme de fréquences et sans besoin que le NES ne soit accordé _a une fréquence spécifique.La dynamique d'un premier type de NES appelé Vibro-Impact NonlinearEnergy Sink (VI-NES) est investiguée expérimentalement grâce à unoscillateur linéaire (OL) à un degré de liberté forcé harmoniquement auquelle VI-NES est attaché. Le pompage énergétique du OL vers le VI-NESest observé expérimentalement, ce qui a permis d'obtenir une importanteréduction du pic de résonance du système principal. Le système est étudiéanalytiquement à l'aide de la méthode Multi-Echelles et le comportementnon-linéaire observé est expliqué théoriquement. Le deuxième type de NES présenté est le Magnetic-Strung NES avec récupération d'énergie. Cette étude ajoute l'aspect lié à la récupération d'énergie au domaine de recherche des absorbeurs non-linéaires. Le système consiste en un oscillateur linéaire (OL) à un degré de liberté forcé harmoniquement auquel le MS-NES est appliqué. La force non-linéaire de rappel peut être modulée grâce à une force magnétique introduite judicieusement, ce qui permet au NES d'avoir plusieurs configurations possibles. Lesystème résultant est un système électromécanique où l'énergie vibratoire dusystème principal est absorbée par le NES et est ensuite dissipée en partiepar l'amortissement visqueux et convertie en partie en puissance électrique.Les études numérique et expérimentale analysent les performances du MSNESen tant qu'absorbeur d'énergie et en tant que récupérateur d'énergie.Finalement, les idées et les perspectives issues de cette étude sont traitéeset les directions pour les travaux futurs sont fournies. / Passive vibration control methods using linear dampers have been largelystudied and investigated, and they have nowadays a broad range of applications.However, linear dampers are efficient when tuned to the specificfrequency to control but present substantial limitations when applied to primarysystems with uncertainties on the modal parameters or to systemshaving a natural frequency that may vary with external forcing.In this thesis the vibration mitigation in mechanical systems by meansof a Nonlinear Energy Sink absorber is studied. The phenomenon governingthe physics of this kind of device is referred to as Targeted Energy Transferand it consists in an irreversible energy transfer from the primary systemto the NES where the energy is then dissipated. This energy transfer mayoccur over a broad range of frequencies with no need for the NES to betuned to a specific one.The dynamics of a first type of NES called Vibro-Impact Nonlinear EnergySink (VI-NES) is experimentally investigated via a harmonically forcedsingle-degree-of-freedom linear oscillator to which a VI-NES is attached. ATargeted Energy Transfer from the LO towards the VI-NES is experimentallyobserved and a significant reduction of the primary system's resonancepeak is obtained. The system is analytically studied by means of the MultipleScales method and the nonlinear behavior experimentally observed istheoretically explained. The second type of NES presented is the Magnetic-Strung NES withenergy harvesting. This study adds the energy harvesting aspect to the researchon nonlinear vibration absorbers. The system consists in a harmonicallyforced single-degree-of-freedom linear oscillator to which the MS-NESis applied. The type of nonlinearity used can be shaped thanks to a magneticforce aptly introduced, allowing the NES to have several possible configurations.The resulting system is an electro-mechanical system in which thevibration energy of the primary system is absorbed by the NES and subsequentlypartially dissipated by the viscous damping and partially convertedinto electrical power. The numerical and experimental studies analyze theperformances of the MS-NES both as an energy absorber and as an energyharvester.

Le travail présenté dans cette thèse est dédié à l’étude d’un absorbeur bistable continu basé sur le principe du "Nonlinear Energy Sink" (NES) et son utilisation pour l’atténuation des vibrations d’un système mécanique à plusieurs degrés de liberté sous excitation acoustique. Le modèle analytique du comportement linéaire de l’absorbeur ainsi que le modèle numérique complet ont été présentés, analysés et validés par des séries d’expériences. Le complexité du transfert énergétique ciblé ("Targeted Energy Transfer" ou TET) entre l'absorbeur et le système primaire à contrôler n’a pas permis une description analytique simple. Nous avons donc choisi de concentrer cette étude sur l’exploration expérimentale et numérique de l’absorbeur couplé à des systèmes mécaniques sous excitations harmonique et aléatoire ainsi que sur l’identification des mécanismes de transfert d’énergie. Le système couplé a montré une dynamique très riche du fait de différents régimes de TET qui ont été décrits dans la littérature pour d’autres types de NES. Ce projet a été financé par Saint-Gobain. L’absorbeur a été adapté pour l’application prévue par la direction industrielle de la thèse: contrôle des vibrations de la double paroi sous excitation acoustique afin d’améliorer l’isolation acoustique fournie par le système.Les connaissances qualitatives sur la dynamique de l’absorbeur obtenues à partir des résultats expérimentaux et numériques, ainsi que l’analogie avec les autres types de NES, ont permis la création d’un absorbeur qui répond à la problématique posée. Les moyens pour l’optimisation et le développement de l’absorbeur ont été identifiés et les simulations préliminaires ont été fournies. / The work presented in this thesis is dedicated to the study of a continuous bistable absorber based on the principle of Nonlinear Energy Sink (NES) and its use for the vibration mitigation of a many-degree-offreedom mechanical systems under acoustic excitation. The analytical model of the linear behavior of the absorber and its complete numerical model were presented, analyzed and validated by series of experiments. The complexity of the Targeted Energy Transfer (TET) between the absorber and the primary system did not allow a simple analytical description. We have chosen to concentrate this study on the experimental and numerical exploration of the absorber coupled to mechanical systems under harmonic and random excitations, as well as on the identification of the mechanisms of energy transfer. The coupled system have shown very rich dynamics as it possessed different regimes of TET, which were earlier described in literature for other types of NES. This project was funded by Saint-Gobain. The absorber was adapted for the application foreseen by the industrial supervisors of the PhD: the vibration control of partitioning double walls under acoustic excitation so that to improve the acoustic isolation provided by the system. The qualitative knowledge on the absorber dynamics obtained from the experimental and numerical results, as well as the analogy with the other types of NES, permitted the creation of an absorber which corresponds to the problematic. The ways for the further optimization and development of the absorber were identified and preliminary simulations were provided.

Lanthanide phosphate (LnPO4) and lanthanide vanadate (LnVO4) nanoparticles (NPs) are promising platforms for theranostic applications because of their chemical stability, low solubility, low toxicity, and unique luminescence and magnetic properties. Motivated by the high radiation resistance and ability to host actinides of naturally occurring lanthanide-based compounds, LnPO4 and LnVO4 NPs were studied as radionuclide carriers for targeted radionuclide therapy using in vivoα-generators, 223Ra, 225Ac, and 227Th. The implementation of these radionuclides has shown potential for the treatment of micrometastases and solid tumors as well as challenges in the retention of decay daughters at the target site to minimize unwanted radiotoxicity. LnPO4 and LnVO4 core-shell NPs doped with either 156Eu, a “cocktail” of 85, 89Sr and 156Eu, or in vivo α-generators 223Ra, 225Ac, and 227Th were synthesized in aqueous media. In vitro retention of radionuclides was investigated by dialyzing the radionuclide-doped NPs suspensions against deionized water and quantifying the activity in dialysate aliquots over time. The crystal structure, morphology, physical stability, luminescence and magnetic properties were evaluated. Partial retention of 156Eu (~70–95%) and 85, 89Sr (>80%) was evidenced in LnPO4 core NPs, while 227Th and decay daughters were quantitatively retained in LaPO4 core + 2 shells NPs (>99%). Gd0.8Eu0.2VO4 and GdVO4 core-shell NPs showed partial retention of 223Ra (~75%), 225Ac (75–95%), 227Th (>96%), and decay daughters. Radionuclide retention was influenced by the lanthanide concentration, crystal structure, and number of shells. The partial retention of radionuclides in both LnPO4 and LnVO4 core-shell NPs may enhance the treatment efficacy while minimizing unwanted toxicity. LnVO4 core and core-shell NPs have potential as carriers of short-lived radionuclides for both diagnostic and therapeutic applications. Emission intensities were higher for LnVO4 with respect to LnPO4 NPs, whereas no significant difference was observed in the magnetic susceptibilities. GdVO4 core NPs displayed enhancement of the signal intensity in T1-weighted images. This work evidences the promising application of both LnPO4 and LnVO4 NPs as platforms for targeted radionuclide therapy and multimodal molecular imaging.

The United States increasingly has resorted to the use of Unmanned Aerial Vehicles (UAVs) for targeted killings of terrorists as a counterterrorism strategy. More states and terrorist organizations also are acquiring UAVs and this development can lead to indiscriminate and unregulated use of UAVs. Previous researchers have indicated the surveillance ability and precise weapon delivery capacity of UAVs make them a weapon of choice for U.S. counterterrorism efforts. Although the U.S. government estimated the collateral damage involved in the use of UAVs at 3-5%, nongovernmental sources put it at 25-40%. A gap exists in the current literature regarding public perception of the use of UAVs as a counterterrorism measure and how international humanitarian law (IHL) may interpret employment of UAVs. The purpose of this quantitative, cross-sectional study is to determine if a relationship exists among public support of the use of UAVs for targeted killing, attitudes towards counterterrorism, and public perceptions of IHL. An online survey was used to collect data from 104 adult participants using the convenience sampling method. Logistic regression, ANOVA, and correlational analyses helped to determine the relationships. The outcomes contributed to the existing literature by providing important data related to public perception of the use of UAVs with the potential to enhance global peace and security. The results contributed to social change initiatives through the potential to facilitate the establishment of international and domestic legal frameworks to regulate the future employment of UAVs for targeted killing.

La prise en charge contemporaine du cancer repose de plus en plus sur le concept de médecine dite personnalisée qui permet, après identification des caractéristiques propres de la tumeur, d'envisager une thérapeutique adaptée à chaque patient. La meilleure connaissance de la biologie des cancers et la mise en évidence de nouvelles cibles thérapeutiques ont ainsi abouti au développement des thérapies ciblées qui vont agir sur une cible cellulaire, généralement surexprimée au niveau d'une tumeur, leur conférant une plus grande spécificité d'action tumorale et un profil d'effets indésirables moins marqué par rapport à la thérapie cytotoxique conventionnelle. Malgré les données d'efficacité clinique, rapportées sur les stratégies thérapeutiques intégrant des thérapies ciblées, on ne dispose pas, malheureusement, des études pharmaco-économiques en cancérologie surtout que ces médicaments se caractérisent par leurs coûts très élevés (exemple: bevacizumab 1000 euros/400 mg ; cetuximab 964 euros/500 mg) et doivent donc être également appréhendés dans le cadre d'études dites en condition de vie réelle. L'objectif de ce projet de recherche est donc de proposer une étude pharmaco- économique sur l'utilisation des thérapies ciblées en cancérologie. Compte tenu du large champ d'investigation, deux affections tumorales distinctes seront considérées : le cancer colorectal métastatique et le glioblastome.Le cancer colorectal fera l'objet d'une étude conduite spécifiquement au Liban et le glioblastome d'une étude en France. L'originalité de cette thèse, conduite dans le cadre d'une coopération franco-libanaise, sera également d'évaluer les impacts médico-économiques de ces nouvelles stratégies selon des perspectives d'organismes payeurs très différentes, ce qui permettra d'appréhender les éventuelles problématiques d'accès aux soins / Although cancer incidence and prevalence are increasing at an alarming rate, progress in the treatment has been slow and treatment benefits are measured in weeks to months. Following the progress in the diagnostic tools and early detection biomarkers, a number of cancer types can be detected before pathological symptoms develops and this is of great significance because individual specific treatment regimens can be designed based on the presence and stage of cancer. In the past decade, there have been considerable improvements in the way that tumors are characterized and knowledge of cancer, at the molecular level has therefore increased greatly and has shift towards the use of targeted cancer therapies that display greater sensitivity and specificity for tumor cells by blocking the activity of the molecule in the host microenvironment that supports tumor growth or inhibiting the activity of protein tyrosine kinases or signaling pathways. Unfortunately, many pharmaco-economic studies are lacking today in the oncologic field. In addition, targeted therapies are very expensive (bevacizumab costs 100 euros/400mg and cetuximab 964 euros/500 mg) and should, therefore, be analyzed in real life conditions. That’s why the main objective of this research-project is to find a pharmaco-economic method on the use of the targeted therapies in oncology. Given the wide scope of investigations, 2 different tumor pathologies will be discussed: metastatic colorectal cancer and glioblastoma. The choice between these 2 cancer types takes place in the growing role of the targeted therapies in the effectiveness and survival of patients as well as their relative costs, especially that they have in common “bevacizumab” targeting the tumor vasculature

Background and purpose. This thesis documents a program of five studies concerned with the assessment of software measurement. The goal of this program is to assist the software industry to improve the information support for managers, analysts and software engineers by providing evidence of where opportunities for improving measurement and analysis exist. Methods. The first study examined the assessment of software measurement frameworks using models of best practice based on performance/success factors. The software measurement frameworks of thirteen organisations were surveyed. The association between a factor and the outcome experienced with the organisations' frameworks was then evaluated. The subsequent studies were more info-centric and investigated using models of information quality to assess the support provided for software processes. For these studies, information quality models targeting specific software processes were developed using practitioner focus groups. The models were instantiated in survey instruments and the responses were analysed to identify opportunities to improve the information support provided. The final study compared the use of two different information quality models for the assessing and improving information support. Assessments of the same quantum of information were made using a targeted model and a generic model. The assessments were then evaluated by an expert panel in order to identify which information quality model was more effective for improvement purposes. Results. The study of performance factors for software measurement frameworks confirmed the association of some factors with success and quantified that association. In particular, it demonstrated the importance of evaluating contextual factors. The conclusion is that factor-based models may be appropriately used for risk analysis and for identifying constraints on measurement performance. Note, however, that a follow-up study showed that some initially successful frameworks subsequently failed. This implied an instability in the dependent variable, success, that could reduce the value of factor-based models for predicting success. The studies of targeted information quality models demonstrated the effectiveness of targeted assessments for identifying improvement opportunities and suggest that they are likely to be more effective for improvement purposes than using generic information quality models. The studies also showed the effectiveness of importance-performance analysis for prioritizing improvement opportunities.

Expressive writing has been linked to positive psychological and health outcomes in general and medical populations, but research examining this intervention in HIV is limited. Higher levels of emotional expression (EE) and depth processing (DP) during writing have been linked to better health status in HIV. Expressive writing has been shown to improve health-related quality of life (HRQoL) in other populations, but has not been examined in HIV. HRQoL is often compromised in HIV+ individuals and therefore improvements in this area are an appropriate goal of psychosocial interventions. This longitudinal study used HLM analyses to examine the relationship between levels of EE and DP during trauma writing and the rate of change in HRQoL over six months in an ethnically diverse sample of 106 HIV+ men and women. Three subscales of the HIV/AIDS-targeted Quality of Life measure were examined: Overall Healthy Functioing (HRQoL-Overall), Without Health Worries (HRQoL-Health), and Life Satisfaction (HRQoL-Life). All longitudinal analyses controlled for demographic (age, gender, race/ethnicity, education), medical (CD4 and VL) and psychological (stressful life events) factors. No significant effects were found for EE/DP to predict changes in HRQoL over time for the full sample. When men and women were examined separately, there was a non-significant tendency for men to decrease in HRQoL over time and for women to increase over time, and a number of EE/DP variables were significant predictors of rate of change in HRQoL. As hypothesized, for women (n = 44) higher level of Experiential Involvement DP predicted greater increase in HRQoL-Overall and HRQoL-Life, and higher negative EE also predicted greater increase in HRQoL-Life over time. Opposite of the direction hypothesized, higher Self Esteem DP predicted a lower level of increase in HRQoL-Life for women. For men (n = 62), findings appeared to be in the opposite direction of women, with greater Self Esteem DP working as a buffer to decreases in HRQoL-Life and HRQoL-Health over time. Furthermore, higher Experiential Involvement and negative EE appeared detrimental for men as both predicted greater decreases in HRQoL-Life over time and Experiential Involvement also predicted greater decreases in HRQoL-Health. Results should be interpreted with caution, as the overall slopes did not show significant change in HRQoL over time. The reasons for observed gender differences are not known. This is the first study to examine the impact of EE and DP in expressive trauma writing on HRQoL in HIV+ individuals. Implications and limitations are discussed.

Quantitative real-time PCR (QPCR) technology has been very useful for diagnosis of viral diseases. QPCR has recently reached a level of sensitivity, simplicity, and reproducibility which allows a large number of samples to be screened rapidly, make it a suitable tool for the clinical virology diagnostics. In this thesis, broadly targeted and degenerated quantitative QPCR assays were used. A somewhat novel single-tube real-time reverse transcription-polymerase chain reaction (QRT-PCR), with takes advantage of ability of rTth DNA polymerase to reverse transcribe RNA in the presence of Mn2+ at elevated temperatures and includes protection against amplimer contamination by using thermolabile UNG, was developed. A new technique for diagnostic of recent viral infection by detection of viral immunoglobulin M (IgM) was also developed. In the first paper, a sensitive single-tube QRT-PCR for detection of enteroviral RNA in patients with aseptic meningitis was presented. In the second paper, a single-serum-dilution real-time PCR-based PIA (PCR-enhanced immunoassay), called quantitative PIA (QPIA), to detect enterovirus IgM for diagnosis of EV infection in patients with aseptic meningitis, was also developed. In the third paper, a broadly targeted, simple, single tube degenerated quantitative QPCR technique for detection of JCV, BKV and SV40 DNA was developed. A conserved region of the VP2 gene of JCV, BKV and SV40 was targeted. A false positive result due to contamination with commonly used SV40 T-antigen plasmids was therefore avoided. In manuscript four, the QPIA assay provide a rational strategy for detection of EV IgM, allows the use of viral antigens isolate from newly diagnosed Type 1 diabetes patients (T1D-EV-QPIA) to measured IgM against diabetogenic viruses in serum from newly diagnosed T1D children, siblings, and healthy children. To conclude, novel broadly targeted real-time PCR methods for diagnosis of entero- and polyoma viral infections were developed.

Severe Traumatic Brain Injury (sTBI) is a major cause of mortality and morbidity. At the Department of Neurosurgery Umeå University Hospital subjects with sTBI are treated with an intracranial pressure (ICP) guided therapy based on physiological principles, aiming to optimise the microcirculation of the brain so avoiding secondary brain injuries. The investigations in this thesis are unique in the sense that all patients with sTBI were treated according to the guidelines of an ICP targeted therapy based on the “Lund concept”. As the treatment is based on normalisation of the ICP, the accuracy and reliability of the measuring device is of outmost importance. Therefore the accuracy, drift, and complications related to the measuring device was prospectively studied (n=128). The drift was 0,9 ± 0,2 mmHg during a mean of 7,2 ± 0,4 days and the accuracy high. No clinical significant complications were noted. In 1997 uni- or bilateral decompressive hemi-craniectomy (DC) was introduced into the treatment guidelines. The effect of DC on the ICP and outcome was retrospectively analysed for subjects with sTBI treated 1998-2001. In the subjects who underwent DC the ICP was 36,4 mmHg immediately before and 12,6 mmHg immediately after the DC. The ICP then levelled out at just above 20 mmHg. The ICP was significant lower during the 72 hours following DC. The outcome did not differ between subjects who had undergone DC or not. Subclinical electroencephalographic seizures and status epilepticus have been reported to be common in subjects treated for traumatic brain injury (TBI). This can negatively influence the outcome giving rise to secondary brain injuries. The occurrence of seizures in subjects treated for TBI using continuous EEG monitoring was therefore prospectively studied. During 7334 hours of EEG recording in 47 patients no electroencephalographic seizures were observed. Theoretically, and based on animal studies, prostacyclin (PGI2) can improve the microcirculation of the brain, decreasing the risk for secondary ischaemic brain injury. PGI2 was introduced to the treatment in a prospective randomised double blinded study (epoprostenol 0,5 ng/kg/min). The effect of PGI1 pkt was analysed using the lactate/pyruvate ratio (L/P) measured by cerebral microdialysis in order to study the energy metabolism in the brain. The outcome was measured as Glasgow Outcome Scale (GOS) at 3 months follow-up. Forty-eight subjects were included. The L/P was pathological high during the first day, thereafter decreasing. There was no significant difference in L/P or outcome between the treated and non-treated group. At 3 months the mortality was 12,5% (95,8% was discharged alive from the ICU), and favourable outcome (GOS 4-5) was 52%. In the same study the brain injury biomarkers S-100B and NSE were followed twice a day for five days to evaluate brain injury and investigate the possible use of these biomarkers for outcome prediction. Initially the biomarkers were elevated to pathological levels which decreased over time. The biomarkers were significant elevated in subjects with Glasgow Coma Scale 3 (GCS) and GOS 1 compared with subjects with GCS 4-8 and GOS 2–5, respectively. A correlation to outcome was found but this correlation could not be used to predict clinical outcome. It is concluded that the ICP measurements are valid and the treatment protocol is a safe and solid protocol, yielding among the best reported results in the world, in regard to favourable outcome as well as in regard to mortality. Epoprostenol in the given dose was not shown to have any effects on the microdialysis parameters nor the clinical outcome. In sTBI L/P and brain injury biomarkers can not be used to predict the final outcome.