Thesis (Ph. D.)--West Virginia University, 2006. / Title from document title page. Document formatted into pages; contains xi, 214 p. : ill. (some col.). Vita. Includes abstract. Includes bibliographical references (p. 155-173).

Th.--Méd.--Nancy 1, 1984. N°: 142.

No description available.

The study explored the nature of the involvement of certain demographic (gender, age and education), cognitive (specifically executive functions) and psychological factors (personality changes and other psychosocial issues) in the disease process in a group of MS patients. A sample of 20 adults (8 male and 12 female) with a mean age of 47,65 years was employed. The assessments procedure entailed a set of neuropsychological measures, a 16 PF questionnaire and a semi-structured interview (with a significant other) from which certain qualitative themes were extracted. The MS sufferers showed deficiencies on measures of executive function, a 16 PF profile in accordance with that of individuals with physical illness and the qualitative themes revolved around issues such as mood, independence, work-status, self-confidence and cognitive difficulties. The data resulting from these assessments supported one another and served to enrich our knowledge regarding the life world of the person with MS. / Psychology / MSc. (Psychology)

L'objectif a été de formuler des émulsions multiples Hydrophile/Lipophile/Hydrophile de taille nanométrique par autoémulsification, d'évaluer leurs cytotoxicités et le taux d'encapsulation. Trois formules ont été développées. La phase lipophile est composée de : triglycéride à chaine moyenne et de deux agents de surface : formules (a) Polysorbate 85 (PS85) / Labrasol®, (b) PS 85 / Cremophor® EL, (c) Glycérol / PS85. Les tailles des globules multiples des formules sont (a) 150 nm, (b) 40 nm et (c) 200 nm. La stabilité est de (a) 24 h, (b) 6 mois et (c) 2 mois. Le système multiple a été caractérisé par microscopie électronique à transmission et calorimétrie différentielle à balayage (DSC). L'évaluation de la viabilité cellulaire a montré une cytotoxicité aigüe (a) et retardé (b) et absence de cytotoxicité (c). La stabilité de la formulation (c) a été améliorée par ajout d'un glycéride hémisynthétique (Suppocire®DM). Différentes méthodes d'évaluation du taux d'encapsulation ont été testées, des méthodes usuelles (ultracentrifugation, dialyse) et d'autres méthodes (dégradation enzymatique, intercalant de l'ADN) avec des résultats montrant une interaction. La mise en évidence de la formation de micelles de PS85 dans la phase aqueuse externe a permis d'expliquer cette interaction. L'ultrafiltration a montré que le PS85 encapsulait du principe actif dans la phase aqueuse externe. L'évaluation de l'encapsulation de DTPA-Europium par DSC a permis détecter un taux d'encapsulation de 68 %. Du DTPA-Gadolinium incorporé dans la phase aqueuse interne a montré une supériorité comme agent de contraste positif par imagerie par résonnance magnétique par rapport à la solution de référence. / The aim was to formulate nanoscale Water/Oil/Water (W/O/W) multiple emulsion (ME) byself-emulsification, to evaluate their cytotoxicity and the encapsulation rate of DTPAEuropiumsolution. Three W/O/W ME have been developed. The lipophilic phase is composed of medium chain triglyceride (MCT) and two surfactants (ratio 7/3), which are : formulations:(a) Polysorbate 85 (PS85) / Labrasol ®, (b) PS85 / Cremophor EL®, (c) Glycerol / PS85. The sizes of multiple droplets are (a) 150 nm, (b) 40 nm and (c) 200 nm. Stabilities are (a) 24 h, (b)6 months and (c) 2 months. Multiple system has been characterized by transmission electronmicroscopy and differential scanning calorimetry (DSC). Evaluation of cell viability showacute cytotoxicity (formulation a), delayed cytotoxicity (formulation b) and no cytotoxicity(formulation c). The stability of formulation (c) has been improved by adding to the oily phase a hemisynthetic glycerid (Suppocire® DM). Different methods of evaluation of encapsulation rate have been tested: conventional methods (ultracentrifugation, dialysis, conductivity) and methods used for other nanosystems (enzymatic degradation, DNA intercalating). Results have shown an interaction. The demonstration of the formation of micelles or an emulsion ofPS85 in the external aqueous phase allowed explaining this interaction. Ultrafiltration was used to characterize it and to show that the PS85 encapsulates the active subtance in the external aqueous phase. Evaluation of the encapsulation of a solution of DTPA - Europium byDSC has overcome this interaction, and detecting an encapsulation rate of about 68 %. The incorporation of a solution of DTPA - Gadolinium in the internal aqueous phase has shown superiority as a positive contrast agent in magnetic resonance imaging as compared to a reference solution.

Mitogen driven differentiation of mononuclear cells is a useful model of antibody synthesis and secretion in humans. We have studied Pokeweed mitogen (PWM) induced immunoglobulin secretion in vitro in both healthy individuals and multiple sclerosis patients. Within the healthy population we have identified individuals who consistently secrete low levels of IgG in response to PWM and others who secrete very high levels. The underlying mechanisms involved in low response are not well understood. We have observed that the peripheral blood mononuclear cells (PBMC) obtained from low responders differ from those obtained from high responders in each of the following: Their T-helper cell subset contains a higher ratio of T suppressor-inducer cells over T helper-inducer cells; their PBMC contain a higher level of in vivo radiation-sensitive suppression; their PBMC generate a lower autologous mixed lymphocyte response; and their B lymphocytes secrete lower amounts of IgG when mixed with heterologous high responder T helper cells. These results suggest the response involves the interactions between T helper cell subsets, T suppressor cells and B lymphocytes and that the level of response is the sum of the contribution of each subset. PWM induced immunoglobulin secretion was measured in multiple sclerosis patients during different phases of clinical disease activity. Relapsing-remitting multiple sclerosis patients in early relapse secreted less immunoglobulin than patients with prolonged relapse, suggesting that immune function varies with clinical disease activity. Testing the level of PWM induced immunoglobulin secretion in relapsing-remitting multiple sclerosis patients during the clinically stable phase suggested that those patients who secreted high levels of IgG in response to PWM were more likely to suffer a clinical relapse within 6 months than those patients who secreted a low amount. Chronic progressive multiple sclerosis patients secreted higher amounts of immunoglobulin in this assay than healthy control individuals. This group of multiple sclerosis patients also had; (i) reduced Concanavalin A (Con A) suppressor cell activity measured both by the ability to suppress a/ Con A induced proliferation and b/ PWM induced IgG secretion in heterologous cell cultures and; (ii) reduced percentages of T cells expressing T suppressor and T suppressor-inducer markers. The treatment of chronic progressive multiple sclerosis patients in vivo with lymphoblastoid interferon resulted in a dramatic reduction in level of PWM induced immunoglobulin secretion without alteration in Concanavalin A induced suppression or in the percentages of T cells expressing subset specific markers. The PWM induced IgG secretion assay is a valuable technique for investigating the regulation of humoral immunity in both health and disease. / Medicine, Faculty of / Pathology and Laboratory Medicine, Department of / Graduate

Le myélome multiple (MM) est une néoplasie B caractérisée par l'accumulation d'un clone plasmocytaire dans la moelle osseuse. Cette pathologie demeure incurable d'où la nécessité d'identifier de nouvelles cibles thérapeutiques. L'utilisation des puces à ADN a permis d'identifier, de nombreux gènes dont l'expression par les cellules de MM est associée à un mauvais ou bon pronostic. La plupart des gènes pronostics identifiés dans le MM codent pour des protéines impliquées dans les processus de réplication, de réparation et de recombinaison de l'ADN. Nous avons voulu aller plus loin dans l'identification et la fonction biologique de ces gènes pronostics. D'une part, nous avons recherché les gènes présentant des pics d'expression très élevés ‘gènes spikés' chez une fraction des patients. Ces gènes sont généralement associés à des évènements oncogéniques. D'autre part, nous avons identifié des gènes pronostics non associés à la machinerie du cycle cellulaire et qui sont fortement exprimés dans des cellules souches pluripotentes ou adultes. L'identification de ces gènes nous a permis de construire un score pronostic très puissant, éliminant les scores pronostics publiés à présent. Un autre aspect majeur est l'élucidation des mécanismes biologiques impliquant ces gènes et qui sont responsables de la résistance aux traitements et/ou de la courte durée de survie des patients, afin de pouvoir les reverser. Nous avons donc évalué le rôle biologique de DEPDC1A un gène fortement exprimée dans les cellules de MM en association avec un mauvais pronostic. Le knockdown conditionnel de DEPDC1A par l'utilisation d'un shRNA, inhibe la croissance des lignées de myélome avec une accumulation des cellules en phase G2/M du cycle cellulaire. Cette accumulation est associée à la phosphosphorylation et à la stabilisation de P53, et à l'accumulation de P21/WAF1. Le knockdown de DEPDC1A résulte également en l'expression de marqueurs de cellules plasmocytaires matures dans les lignées de MM : CD31, CD38, CD138, IL6R, CXCR4, CD9, VLA6. DEPDC1A contrôle donc le cycle cellulaire des plasmocytes tumoraux en interférant avec la voie P53 et bloque leur différenciation. Ces travaux montrent que DEPDC1A pourrait jouer un rôle essentiel dans la croissance des cellules de myélome et pourrait être une cible moléculaire prometteuse pour de nouvelles drogues ou de peptides-vaccins dans le MM. / Multiple myeloma (MM) is a B neoplasia characterized by the accumulation of a plasma cell clone in the bone marrow. This disease remains incurable, hence the need to identify new therapeutic targets. The use of DNA microarrays has identified many genes whose expression in MM cells is associated with poor or good prognosis. Most of the genes identified in the MM predictions encode proteins involved in DNA replication, repair and recombinaison processes. We wanted to go further in the identification and biological function of these prognostic genes.First, we looked for genes that have a spike expression, i.e. they are highly expressed in MMCs of a fraction of patients. These genes are generally associated with oncogenic events.On the other hand, we have identified pluripotent and adult stem cell genes unrelated to cell cycle and aberrantly expressed by human multiple myeloma cells in association with poor prognosis. The identification of these genes has allowed us to build a powerful prognostic score, stonger than already published scores.Another major aspect is the elucidation of biological mechanisms involving these genes that are responsible for drug resistance and/or short-term survival of patients, to revert them. We evaluated the biological role of DEPDC1A gene which are highly expressed in MM cells in association with a poor prognosis. The conditional knockdown of DEPDC1A by using an shRNA, inhibits the growth of myeloma cell lines with an accumulation of cells in G2/M phase of cell cycle. This accumulation is associated with phosphosphorylation and stabilization of p53, and accumulation of P21/WAF1. The knockdown of DEPDC1A also results in the expression of mature plasma cell in MM cell lines: CD31, CD38, CD138, IL6R, CXCR4, CD9, VLA6. DEPDC1A therefore controls the cell cycle of plasma cells by interfering with the p53 pathway and blocks their differentiation. This work shows that DEPDC1A could play a role in the growth of myeloma cells and could be a promising molecular target for new drugs or vaccine peptides in MM.

Le myélome multiple (MM) est une néoplasie lymphocytaire B qui se caractérise par l'accumulation d'un clone de cellules plasmocytaires tumorales dans la moelle osseuse interagissant de manière étroite avec le microenvironnement. L'étude du profil d'expression génique à l'aide des puces à ADN a permis de préciser l'hétérogénéité de cette pathologie et de découvrir de nouveaux acteurs susceptibles d'avoir un rôle importent dans sa physiopathologie. La surexpression de Oct-3/4, Sox2, c-Myc et KLF4 dans des cellules adultes les reprogramme à l'état de cellules souches. J'ai montré une surexpression significative de 3 de ces 4 gènes – KLF4, MYC, SOX2 - dans le MM. Ces gènes sont également fréquemment surexprimés dans 18 types de cancers sur 40 étudiés. Par ailleurs, leur expression peut y être associé à un mauvais pronostique ou un signe de progression tumorale, dus peut-être à leur capacité à induire des caractéristiques de cellules souches cancéreuses. Nous avons par la suite débuté l'étude de la fonction des protéines codées par ces gènes dans le MM, en commençant par KLF4, facteur de transcription activateur ou répresseur. KLF4 est exprimé dans les plasmocytes (PC) normaux, mais son expression est perdue dans les PC tumoraux (cellules myélomateuses, MMC) de 2 patients sur 3 atteints de MM au diagnostic. Parmi les patients pour lesquels les MMC expriment KLF4, se trouve un groupe de patients à haut risque, le groupe MMSET portant la translocation t(4 ;14). L'utilisation d'un modèle inductible d'expression de KLF4 dans des lignées de MM avec transduction lentivirale, a mis en évidence un arrêt du cycle associé à l'expression de P27/KIP1 pour les lignées avec des mutations inactivatrices de P53, mais également de P21/WAF1 pour des lignées avec P53 sauvage. Cette sortie du cycle due à l'expression de KLF4 pourrait protéger les plasmocytes tumoraux de l'apoptose induite par certaines drogues ciblant le cycle, comme nous l'observons in vitro avec le Melphalan.L'objectif majeur de notre équipe est de comprendre le fonctionnement du PC normal et celui du PC tumoral. Afin d'atteindre cet objectif, il est nécessaire d'obtenir un système efficace permettant d'introduire un gène dans un PC. Nous avons montré que des lentivirus pseudotypés avec des glycoprotéines tronquées (hemagglutinine et fusion) issus du virus de la rougeole, transduisent de façon stable et efficace des PC normaux et tumoraux. / Multiple myeloma (MM) is a B-cell neoplasia characterized by the accumulation of a clone of malignant plasma cells in bone marrow closely interacting with its microenvironment.Gene expression profiling using DNA microarrays has clarified the heterogeneity of this disease and has allowed the finding of new actors that may have an important function in MM pathophysiology.Overexpression of Oct-3/4, Sox2, c-Myc and KLF4 in adult cells causes their return to the state of stem cells, commonly called induced pluripotent stem cells (iPS). Our team has shown a significant overexpression of at least one of these four factors in 18 out of 40 cancers studied. Moreover, their expression may be associated with poor prognosis or may be a sign of tumor progression, perhaps due to their ability to induce characteristics of cancer stem cells.We therefore began the study of the function of these genes in MM, starting with KLF4, which can either be an activator or a repressor of transcription, depending on the promoter. KLF4 is expressed in normal plasma cells (PC), but its expression is lost in 2 out of 3 patients with MM at diagnosis. Among patients for whom the PCs express KLF4, is a group of high-risk patients, the MMSET group, bearing the t(4;14) translocation.An inducible model of KLF4's expression in MM cell lines was obtained using lentiviral transduction. Our model revealed a KLF4 induced cycle arrest, associated with the expression of P27/KIP1 when P53 is mutated, but also P21/WAF1 in case of wild type P53. This cell cycle blockade due to the expression of KLF4 could protect malignant plasma cells from the apoptosis induced by certain drugs targeting the cell cycle, as shown by our in vitro observations using melphalan.The main goal of our team is to understand the normal function of PCs and the PC tumor. To achieve this, it is necessary to obtain an effective system for introducing a gene in a given PC. We have shown that lentiviruses pseudotyped with truncated glycoproteins (Hemagglutinin and Fusion) from measles virus, can a stably and efficiently transduce normal and malignant PCs.

An in vitro IgG secretion assay was developed to investigate the regulation of the humoral immune response in humans. Pokeweed mitogen (PWM), a plant lectin derived from Phytolacca americana stimulates human peripheral blood mononuclear cells (PBMNC) to divide and resting B-lymphocytes to differentiate into immunoglobulin secreting cells (ISC). This differentiation requires that both monocytes and T-lymphocytes be present in the culture system. The amount of IgG secreted by these differentiated B-lymphocytes in response to PWM appears to be the net result of a balance between the functional activity of the regulatory T-helper and T-suppressor cells. Alterations, qualitative or quantitative in any of these leukocyte subsets could conceivably alter the amount of IgG secreted by the B-lymphocyte subpopulation. We have employed this assay to investigate the immune status in a group of chronic progressive multiple sclerosis (MS) patients and to assess the immunoregulatory effects of interferon (Wellferon R, INF) administered in vivo to this selected group. Their mononuclear cells (MNC) were studied in this PWM induced IgG secretion assay before INF treatment and again after 7 days of daily sub-cutaneous injections (5 X 10⁶ u/day). Twenty patients received the interferon (INF) preparation and eighteen received normal saline. The study was carried out in a double blind manner and the code was broken only after individual results had been calculated. / Medicine, Faculty of / Pathology and Laboratory Medicine, Department of / Graduate

A high level of “environmental awareness” in the participating countries will likely raise the success of the Kyoto Protocol negotiated in Paris in 2015. In this context it is of interest to investigate the diffusion of environmental awareness, and also the (economic) factors, on which this diffusion depends. This paper addresses these questions for the regions of the Russian Federation, which are sufficiently diverse regarding cultural and economic issues. From a formal point of view, a “Multiple-Indicator-Multiple- Causes” (MIMIC) approach, based on a variety of “indicators” for environmental awareness, derived from search entries in c Yandex, and a variety of “causes”, economic and socio-economic factors, is applied. The empirical results point first to a strong dependence of environmental awareness on the level of GRP per capita. Moreover, the diffusion seems to spread from the eastern part of Russia towards the western regions.