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1
Sutha, Ken. "Osteoinductive material derived from differentiating embryonic stem cells". Diss., Georgia Institute of Technology, 2012. http://hdl.handle.net/1853/51722.
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The loss of regenerative capacity of bone, from fetal to adult to aged animals, has been attributed not only to a decline in the function of cells involved in bone formation but also to alterations in the bone microenvironment that occur through development and aging, including extracellular matrix (ECM) composition and growth/trophic factor content. In the development of novel treatments for bone repair, one potential therapeutic goal is the restoration of a more regenerative microenvironment, as found during embryonic development. One approach to creating such a microenvironment is through the use of stem cells. In addition to serving as a differentiated cell source, pluripotent stem cells, such as embryonic stem cells (ESCs), may possess the unique potential to modulate tissue environments via local production of ECM and growth factors. ESC-produced factors may be harnessed and delivered to promote functional tissue regeneration. Such an approach to generate a naturally derived, acelluar therapy has been employed successfully to deliver osteoinductive factors found within adult bone, in the form of demineralized bone matrix (DBM), but the development of treatments derived instead from developing, more regenerative tissues or cells remains attractive. Furthermore, the derivation of regenerative materials from an ESC source also presents the added benefit of eliminating donor to donor variability of adult, cadaveric tissue derived materials, such as DBM. Thus, the objective of this project was to examine the osteoinductive potential harbored within the embryonic microenvironment, in vitro and in vivo. The osteogenic differentiation of mouse ESCs as embryoid bodies (EBs) was evaluated in response to phosphate treatment, in vitro, including osteoinductive growth factor production. The osteoinductivity of EB-derived material (EBM) was then compared to that of adult tissue-derived DBM, in vivo. Phosphate treatment enhanced osteogenic differentiation of EBs. EBM derived from phosphate treated EBs retained bioactive, osteoinductive factors and induced new bone formation, demonstrating that the microenvironment within osteogenic EBs can be harnessed in an acellular material to yield in vivo osteoinductivity. This work not only provides new insights into the dynamic microenvironments of differentiating stem cells but also establishes an approach for the development of an ESC-derived, tissue specific therapy.
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Dashottar, Amitabh. "Posterior Shoulder Tightness Measurements: Differentiating Capsule, Muscle and Bone". The Ohio State University, 2012. http://rave.ohiolink.edu/etdc/view?acc_num=osu1337880690.
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Danilewitz, Larry Mark. "A phenomenological investigation into the psychoanalytic psychotherapist's experience of identifying, differentiating and processing the patient's transference-based and reality-oriented reactions". Thesis, Rhodes University, 1993. http://hdl.handle.net/10962/d1002469.
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The aim of this study was to describe the psychoanalytically-oriented therapist's experience of identifying, differentiating and processing the patient's transference-based and reality-oriented reactions. In order to investigate the therapist's lived experience of being receptive to the total communication of the patient in the analytic situation, the researcher adopted the empirical phenomenological method. This descriptive and intuitive method grounded the researcher in the concreteness of the everyday life-world of the therapist, and enabled him to explicate the therapist's immediate, pre-theoretical experiences of his patient. The appropriate central research question, formulated to elicit the experience of this phenomenon, emerged through the process of enquiry during the pilot study. Thirteen experienced, psychoanalytically-oriented psychotherapists were interviewed and the five protocols considered most revelatory of the phenomenon under investigation were analyzed in detail. The remaining eight protocols were used to illuminate central themes and to clarify areas of uncertainty during the phase of formal explication. The central findings revealed that the oscillating process of the therapist as he shifts from being immersed in the world of his patient to being in a position of observation and self reflection is the fulcrum around which he evaluates the nature of his patient's communications. During this ongoing process of discrimination, living in duality, the therapist comes to experience himself as a patient scrutinized by his own and his patient's confrontations. His journey of disentanglement, the endeavour to differentiate his responses from his patient's actions, is dependent on his ability to engage in honest selfreflection and to access his pre-theoretical and articulated cognitions of his patient. This allows him to acknowledge his own role in what has unfolded interpersonally and to appropriate his previously denied feelings for and attitudes towards his patient, a prerequisite for the accurate and full appraisal of the nature of his patient's communications. Forsaking fixed judgements, the therapist becomes open to the confluence between the reality-oriented responses and transference-based reactions of his patient. This salient discovery, when dialogued with the literature, reinforced the theories of Greenson and Langs that not all the interactions between the patient and the analyst/therapist are transference-based and that it is therefore imperative that the analyst/therapist reflect on his participation in the analytic situation.
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Knerr, Michael R. "Differentiation and Power in Couples Therapy". The Ohio State University, 2008. http://rave.ohiolink.edu/etdc/view?acc_num=osu1221759872.
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Queron, Brenda. "Étude du mode d'action du DV188 dans l'inhibition des propriétés souches et tumorigéniques des cellules souches cancéreuses de gliome". Electronic Thesis or Diss., Université Côte d'Azur, 2024. http://www.theses.fr/2024COAZ6050.
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Les glioblastomes se classent parmi les tumeurs cérébrales les plus agressives, caractérisées par une hétérogénéité intratumorale, une faible espérance de vie et une résistance prononcée aux traitements par radio-chimiothérapie. La complexité de ces tumeurs réside notamment dans la présence de cellules souches de gliomes (CSG), impliquées dans la prolifération clonale, l'invasivité et la récidive tumorale. Les CSG expriment divers marqueurs de cellules souches et de pluripotence tels que NANOG et SOX2. Ces facteurs de transcription doivent être transloqués dans le noyau cellulaire pour activer desprogrammes géniques qui maintiennent les propriétés souches et la tumorigénicité des CSG. Le traitement conventionnel consiste en une chirurgie, lorsque cela est possible, suivie d'une radiochimiothérapie. Malheureusement, ce traitement demeure limité, et les tumeurs récidivent en raison dela sélection et de la persistance des CSG résistantes après l'arrêt du traitement. Pour surmonter ce phénomène dû à la pression cytotoxique, nous avons cherché à élaborer une nouvelle stratégie thérapeutique visant à induire la différentiation des CSG en un phénotype moins agressif et plus sensible au traitement conventionnel. Dans ce contexte, nous avons identifié un composé chimique, le DV188, issu d'une bibliothèque de molécules synthétisée par l'Institut de Chimie de Nice. Nos résultats ont montré que ce composé est capable d'induire la différentiation des CSG dérivées de patients, d’inhiberleur capacité de prolifération clonale et de les sensibiliser à l’agent chimiothérapeutique de référence, le temozolomide (TMZ). Plus important encore, le DV188 empêche in vivo l'initiation et la progression tumorale sans affecter la survie de la souris après des mois de traitement. De plus, la combinaison de traitement du DV188 et du TMZ a démontré une efficacité deux fois supérieure à celle du TMZ seul. Au niveau moléculaire, nous avons mis en évidence un effet du DV188 sur le transport nucléaire de facteurs essentiels au maintien de l'état souche, perturbant ainsi les mécanismes favorisant l'agressivité et la tumorigénicité des CSG. Ces données soutiennent fortement l'idée que le ciblage de la différentiation des CSG représente une voie thérapeutique prometteuse contre le glioblastome. Dans la lignée de cette avancée, et compte tenu de l'efficacité démontrée du DV188 dans les modèles murins, l'exploration de sa combinaison avec l'agent chimiothérapeutique de référence s'impose comme une nouvelle stratégie synergique potentielle pour le traitement de cette maladie dévastatriceGlioblastomas are the most aggressive brain tumors, characterized by intratumoral heterogeneity, poor life expectancy, and significant resistance to radio-chemotherapy treatments. The complexity of these tumors is further exacerbated by the presence of glioma stem cells (GSC), which play a crucial role in clonal proliferation, invasiveness, and tumor recurrence. GSC express various stem cell and pluripotency markers, such as NANOG and SOX2. These transcription factors must be translocated into the nucleus to activate gene programs that maintain stemness and tumorigenic properties of GSCs. The conventional treatment involves surgical intervention, when possible, followed by radio- chemotherapy. Unfortunately, this conventional treatment is limited, and tumors relapse due to the selection and the persistence of resistant GSC upon treatment cessation. To overcome this issue resulting from cytotoxic pressure, we aimed to develop a novel therapeutic strategy based on the differentiation of GSC into a less aggressive phenotype that is more sensitive to conventional treatments. In this context, we identified a chemical compound, DV188, synthesized from a molecule library by the Institute of Chemistry in Nice. Our results indicate that this compound effectively induces differentiation of patient-derived GSC, inhibits their clonal proliferation capacity, and sensitizes them to the standard chemotherapeutic agent, temozolomide (TMZ). Importantly, DV188 prevents in vivo tumor initiation and progression without affecting mouse survival following months of treatment. Additionally, the combination of DV188 and TMZ treatment demonstrated twice the efficacy compared to TMZ alone. At the molecular level, we identified an effect of DV188 on the nuclear transport of essential factors required for maintaining stem cell properties, thereby disrupting mechanisms that drive GSC aggressiveness and tumorigenicity. Our data strongly support the idea that targeting GSC differentiation, through the inhibition of nuclear transport of transcription factors involved in maintaining stemness properties, represents a promising therapeutic avenue against glioblastoma. In line with this advancement and considering the demonstrated efficacy of DV188 in murine models, the exploration of its combination with the standard chemotherapeutic agent emerges as a potential new synergistic strategy for treating this devastating disease
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Enane, Francis Obunyakha. "HEPATOCYTE DIFFERENTIATION AND HEPATOCELLULAR CARCINOMA: RATIONALE FOR P53 INDEPENDENT THERAPY". Case Western Reserve University School of Graduate Studies / OhioLINK, 2017. http://rave.ohiolink.edu/etdc/view?acc_num=case1491570319727552.
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Yang, Ya-Ting. "Molecularly targeted therapy for ovarian cancer". Columbus, Ohio : Ohio State University, 2006. http://rave.ohiolink.edu/etdc/view?acc%5Fnum=osu1149015359.
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EL, SAID DALYA. "Blood derived stem cells (BDSCs): neural differentiation protocols for human therapy". Doctoral thesis, Università degli Studi di Roma "Tor Vergata", 2012. http://hdl.handle.net/2108/209984.
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La tecnologia delle cellule staminali hanno causato un notevole entusiasmo tra gli interessati alla salute degli animali e degli uomini. Molte persone stanno cercando nuove cure per le malattie e la terapia i vivo. Noi sappiamo che tutti i tipi di cellule possono essere rigenerate eccetto le cellule neuronali di mammifero, sebbene il tessuto nervoso periferico si rigeneri la guaina mielinica che permette l’orientamento della fibra. L’importanza di ottenere cellule neuronali funzionali è vitale per le malattie neurodegenerative. Le malattie neurodegenerative sono per definizione delle malattie progressive corniche caratterizzate da una selettiva perdita di neuroni in aree motori simmetriche, sensoriali e cognitive appartenenti al SNC, o perdita/ disfunzione delle fibre mieliniche o non mieliniche nel SNP. Oggi è possibile deprogrammare cellule adulte in cellule staminali pluripotenti in vitro utilizzando specifici protocolli e sostanze non tossiche per l’uomo, ottenere neurosfere (corpi cellulari da dove è possibile isolare e studiare le cellule staminali neuronali). In questo lavoro di tesi ho dimostrato come è possibile utilizzando cellule deprogrammate del sangue ottenere neurosfere e differenziarle in neuroni adulti in vitro e come questi miei risultati siano in accordo con i risultati ottenuti in vivo.Stem cells technology has provoked considerable excitement among people interested in welfare of animals and humans. Many people are searching for new treatments of diseases and in vivo therapy. We know that all types of cells can be regenerated except neural cells of mammals such as don’t regenerate, although the peripheral nervous tissue regenerate if neurillematic sheath allow the orientation of fiber. the importance of obtaining functional nerve cells is vital in neurodegenerative diseases. Neurodegenerative diseases are by definition progressive chronic diseases characterized by a selective loss of neurons in areas, symmetric motor , sensory , cognitive and member ship of CNS, or loss / dysfunction of myelinated or non myelinated fibers in the PNS. It has been possible today to use BDSCs after Deprogrammation of adult peripheral blood, and using specific protocols to obtain “neurospheres” from these cells in specific medium and non toxic substances addition. Neurospheres (composed of neural stem cells) provide a method for investigating neural precursor in vitro instead of embryonic stem cells. In this study I have demonstrated how BDSCs being able to form neurospheres and differentiated into mature neurons in vitro, and how it is possible to use stem cells therapy as treatment in vivo.
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Harrison, Adrian Paul. "Regulation of porcine skeletal muscle growth and differentiation". Thesis, University of Cambridge, 1995. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.360936.
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Razvadauskaite, Giedre. "Survival and differentiation of implanted skeletal myoblasts in the native and in the cryoinjured myocardium". Link to electronic thesis, 2003. http://www.wpi.edu/Pubs/ETD/Available/etd-0106103-155714.
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Thesis (M.S.)--Worcester Polytechnic Institute.Keywords: myoblasts; dexamethasone; infarction; cryoinjury; desmin; myosin heavy chain; differentiation. Includes bibliographical references (p. 54-59).
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Hu, Zhengqing. "Investigating a cell replacement therapy in the inner ear /". Stockholm, 2004. http://diss.kib.ki.se/2005/91-7140-170-9/.
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Holmström, Niklas. "Directed differentiation of adult neural stem cells for cell therapy in the nervous system /". Stockholm, 2005. http://diss.kib.ki.se/2005/91-7140-358-2/.
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Corti, Serena. "Uncoupling tumorigenicity from dopaminergic differentiation potential of hiPSCs by acting on Glypican4 : combining in vitro differentiation studies with preclinical studies for Parkinson's disease therapy". Thesis, Aix-Marseille, 2019. http://www.theses.fr/2019AIXM0439.
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Les cellules souches pluripotentes induites humaines (hiPSC) constituent une avancée décisive pour la science biomédicale, car elles peuvent générer un nombre illimité de cellules pour la thérapie cellulaire (TC). La différentiation de neurones dopaminergiques (DA) à partir d’hiPSC est très prometteuse pour la TC dans la maladie de Parkinson (MP), mais les protocoles de différenciation ne sont pas optimisés: l'efficacité de différenciation varie et des SC non différenciés peuvent provoquer une tumeur lors d'une transplantation. J'ai étudié la possibilité d'augmenter la différenciation d’hiPSC en neurones DA, en modulant la perception de la signalisation cellulaire, par la régulation négative du héparan-sulfate protéoglycane Glypican4 (GPC4). J'ai observé que la régulation négative de GPC4 est une stratégie efficace pour augmenter la différenciation DA, comme l'indique l'augmentation des marqueurs DA in vitro et in vivo après transplantation dans le striatum de rats modèles pour la MP. De plus, dans les cellules GPC4-mut les marqueurs de cellules souches neurales étaient diminués ou identiques aux contrôles et des voies alternatives étaient dérèglées. Enfin, suite à une xénogreffe chez des souris, j’ai observé que la régulation négative de GPC4 réduit la formation de tumeurs. Cette altération du potentiel tumorigène pourrait réduire les risques de tumeurs dans les TC. L’ensemble des résultats de cette thèse fournissent de nouvelles informations sur la génération de cultures DA, ainsi que des protocoles de différenciation plus standardisés, qui peuvent être utilisés pour augmenter l’efficacité, le rendement et la qualité des produits thérapeutiques dérivés de hiPSCHuman induced pluripotent stem cells (hiPSCs) are a breakthrough for biomedical science, as they can generate an unlimited number of cells for cell replacement therapy (CRT). The derivation of dopaminergic (DA) neurons from hiPSCs is very promising for CRT in Parkinson´s disease (PD). Nevertheless, differentiation protocols are not optimal: the differentiation efficiency varies in different experiments and persisting SCs can cause tumour growth after transplantation. I investigated the possibility of increasing hiPSC differentiation into DA neurons by modulating cell signalling perception through Glypican4 (GPC4) downregulation. GPC4 is a heparan sulphate proteoglycan that is crucial for fine-tune perception of extracellular cues. I observed that GPC4 downregulation is an efficient strategy to increase specifically DA differentiation, as assessed by upregulation of DA markers in GPC4-mutant cells both in vitro and in vitro after transplantation into the striatum of PD rats. Moreover, in GPC4-mut line, neural SC markers were downregulated or equally expressed and alternative fates were disadvantaged in vitro. Finally, by performing flank xenograft in nude nice, I observed that GPC4 downregulation can also impair tumor development, thus revealing a potential relevance for hiPSC-based CRT. Generally, the results of this thesis provide new insights into generation of standardized vmDA cultures, which can be used to increase efficiency, throughput scale and quality of hiPSC-derived therapeutic products
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Salvatico, Jose. "The Expression of MKRN1, an E3 Ubiquitin Ligase for Telomerase Reverse Transcriptase, Is Induced with Differentiation Therapy in Leukemia". Master's thesis, University of Central Florida, 2009. http://digital.library.ucf.edu/cdm/ref/collection/ETD/id/3744.
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Telomeres are important structural and functional components of chromosomes, serving to provide stability and enabling full replication of the chromosomes. However, a shortening of the telomeres occurs with each cell division that can be fixed by a polymerase activity provided by telomerase, preventing this loss which would otherwise eventually lead to chromosome end-to-end fusions, senescence and cell death. The telomerase activity is present in stem cells and germ line cells, but absent or barely noticeable in adult somatic cells. However, in approximately 80-90% of transformed somatic cells the telomerase activity is recovered, resulting in a "telomerase positive phenotype". This phenotype has been a prime target in cancer research, and recently a novel mechanism for regulating telomerase levels has been uncovered. Makorin 1 RING finger protein (MKRN1) was found to be an E3 ubiquitin ligase for hTERT, the rate-limiting catalytic component of telomerase, leading to the ubiqutin-mediated 26s proteasomal degradation of hTERT and reduced telomerase activity. So, MKRN1 plays a role in telomere homeostasis. In this study we looked at the expression of MKRN1 in numerous tumor cell lines (Hela, HCT116, HL60) and the normal diploid fibroblasts (WI-38). In the latter cell line, basal levels of MKRN1 were found to increase 6-fold when the cells were serum starved and arrested in G1/G0. In contrast, the cancer cell lines expressed MKRN1 at low levels or undetectable. This would indicate that MKRN1 is up-regulated in resting or G1 arrested cells.In one cell line the promyelocytic leukemia, HL-60, showed no protein levels of MKRN1. This cell line is able to be terminally differentiated upon ATRA treatment, when cells are arrested at G1. In this model system of cellular differentiation hTERT mRNA levels and telomerase activity decrease drastically and quickly. We hypothesized that the differentiation of HL-60 induced by ATRA would be accompanied by an increase in MKRN1 levels. MKRN1 mRNA and protein levels were strongly up-regulated during the ATRA-mediated differentiation of HL-60 cells. Although, a decrease in hTERT mRNA is a contributor to telomerase inhibition during cellular differentiation; our data indicate that the up-regulation of MKRN1 ensures the effective removal of residual telomerase activity by the ubiquitin-mediated degradation pathway at the proteasome.M.S.
Department of Molecular Biology and Microbiology
Burnett College of Biomedical Sciences
Molecular and Microbiology MS
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Britschgi, Adrian. "DAPK2, EGCG and autophagy in neutrophil differentiation and retinoic acid-therapy of acute myeloid leukemic cells". [S.l.] : [s.n.], 2009. http://www.zb.unibe.ch/download/eldiss/09britschgi_a.pdf.
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Dodhy, Asad. "Population based evaluation of actin cytoskeletal morphometric descriptors as characterisation of stem cell differentiation". Thesis, Queen Mary, University of London, 2018. http://qmro.qmul.ac.uk/xmlui/handle/123456789/46030.
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Stem cells have yet to contribute to their full potential in the field of regenerative medicine and further understanding of the underlying kinetics of cell differentiation could be the step forward. Various methods have been used to characterise stem cell lineage commitment. However, most of these techniques are end-point assays and provide very little information about the changes occurring in the early stages of the differentiation process. This project aims to explore if the structural and geometrical specificity of the cytoskeletal components (actin in particular) encode information regarding cell lineage. Adipogenic and osteogenic differentiation lineages were selected, as they have been extensively studied over the past few decades. We have developed a novel approach to describe cells by defining their cytoskeletal and nuclear morphology in terms of 19 geometric measurements. This set of parameters has a range of complexity, extending from one dimensional (e.g. fibre length, fibre thickness) to compound geometrical readings (e.g. chirality and fibre alignment), while some estimate morphological and mechanical properties of the nucleus i.e. Poisson ratio and chromatin condensation. A proprietary image analysis algorithm is used to analyse fluorescent images of cells biochemically and mechanically stimulated to differentiate for a period of up to 10 days. Our analysis pipeline is currently optimised for images acquired at x20 magnification using epi-fluorescence but can be further adapted for high throughput live cell imaging. Factorial analysis of the measured features showed that some parameters change markedly in the early stages of differentiation. More interestingly we observed these changes to be non-linear and non-monotonic. This analysis, in light with previously published literature on the subject has allowed us to more intricately hypothesise probable mechanisms involved with mechanotransduction which direct the lineage commitments. As our technique quantifies the morphology of individual cells, we used our extracted feature data to characterise each cell using a multivariate predictive model (LDA).
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GAMBINI, ELISA. "Isolation, characterization and ex vivo amplification of heart-derived c-kit progenitor cells for cellular therapy". Doctoral thesis, Università degli Studi di Milano-Bicocca, 2009. http://hdl.handle.net/10281/7781.
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The myocardium contains a resident progenitor cells, named cardiac progenitor cells, that maintain the heart homeostasis by replacing lost myocytes. The c-kit tyrosine kinase receptor is one of the principal markers defining cardiac progenitors in the adult mammalian heart, including human. C-kit+ cardiac progenitors have been recently identified as a multipotent cells population able to give rise to myocardial, endothelial and smooth muscle both in vitro and in vivo. The discovery of cardiac progenitors and the devise of culture systems to amplify these cells in culture without loosing their differentiation potency has opened new perspectives for clinical applications aimed at replacing myocardium lost as a consequence of acute or chronic ischemic disease. While cardiac progenitors have been deeply investigated for their stem cell activity, susceptibility to stress conditions and aging, and their ability to repair the infarcted heart, their origin, phenotype and biological features have been disclosed only in part. In the present study, we report that c-kit+ cardiac progenitors derived from the human heart express markers in common with mesenchymal stem cells. Our in vitro analyses revealed that, compared to bone marrow-derived mesenchymal cells, c-kit+ cardiac progenitors have a reduced ability to differentiate into canonical mesenchymal cellular derivatives such as osteoblasts and adipocytes; by contrast, under appropriate differentiation conditions, these cells gave rise to tubular-like structures at the same extent as endothelial cells, expressed smooth muscle cells markers, and expressed early and late myocardial markers. These findings suggest that ex-vivo amplified c-kit+ cardiac progenitors differentiate into mesenchymal cells having a preferentially myocardial commitment.
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DiVincenzo, Lola S. "DIRECTION OF INDUCED PLURIPOTENT STEM CELL DIFFERENTIATION BY ENDOTHELIAL CELL SECRETOME". Kent State University / OhioLINK, 2015. http://rave.ohiolink.edu/etdc/view?acc_num=kent1438031276.
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Hoffmann, Sebastian Benedikt [Verfasser], e Ralf [Akademischer Betreuer] Wagner. "In vitro differentiation of genome engineered stem cells for a novel HIV therapy / Sebastian Benedikt Hoffmann ; Betreuer: Ralf Wagner". Regensburg : Universitätsbibliothek Regensburg, 2019. http://d-nb.info/1200208986/34.
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Farouz, Yohan. "Designing biomaterials for controlled cardiac stem cell differentiation and enhanced cell therapy in the treatment of congestive heart failure". Thesis, Sorbonne Paris Cité, 2015. http://www.theses.fr/2015USPCB114/document.
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La thérapie cellulaire se positionne comme une stratégie prometteuse pour inciter le cœur infarci à se régénérer. A cet effet, des études récentes placent des espoirs considérables dans l’utilisation des cellules souches embryonnaires et notre laboratoire a déjà démontré comment les différencier en progéniteurs cardiovasculaires, un type de précurseurs cellulaires qui ne peut aboutir qu’à la formation de cardiomyocytes, de cellules endothéliales ou de cellules de muscles lisses. Cet engagement précoce réduit leur capacité de prolifération anarchique et en même temps leur permet de rester suffisamment plastiques pour éventuellement s’intégrer plus facilement avec le tissue hôte. Cependant, les études précliniques et cliniques d’injection de ces cellules s’avérèrent décevantes. Malgré de légères améliorations de la fonction cardiaque, on observa une trop faible survie cellulaire ainsi qu’un taux de rétention des cellules dans le myocarde remarquablement bas. Afin d’étudier ce problème, mes travaux de thèse ont porté non seulement sur la conception de nouveaux biomatériaux pouvant servir de moyen de transport et d’intégration des cellules dans la zone infarcie, mais aussi sur la conception de biomatériaux permettant de contrôler précisément l’environnement cellulaire au cours du processus de différenciation de cellules souches pluripotentes humaines en cardiomyocytes. Grâce aux importantes interactions entre nos laboratoires de recherche fondamentale et de recherche clinique, nous avons tout d’abord développé de nouvelles techniques de fabrication et de caractérisation de patches de fibrine cellularisés qui sont récemment entrés dans un essai clinique de phase I. A partir de cette formulation clinique approuvée par les autorités de régulation, nous avons élaboré toute une gamme de matériaux composites uniquement à base de matières premières pertinentes dans ce cadre clinique, dans le but d’améliorer la maturation des progéniteurs cardiovasculaires une fois greffés sur le cœur défaillant. Dans cette optique, nous avons également développé un modèle in vitro permettant d’étudier précisément l’influence combinée de la rigidité du substrat et du confinement spatial sur la différenciation des cellules souches en cardiomyocytes. Grâce à des techniques de microfabrication sur substrat mou, il a été possible de positionner précisément les cellules souches pluripotentes dans des espaces restreints d’élasticité variable. Ainsi, nous avons pu observer que même en utilisant des protocoles chimiques éprouvés basés sur la modulation de cascades de signalisation impliquées dans le développement cardiaque, une très forte hétérogénéité pouvait apparaître en fonction de l’environnement physique des cellules. Nous avons ainsi pu extraire les caractéristiques principales permettant une différenciation cardiaque efficace, reproductible et standardisée et les avons appliquées à la fabrication d’une nouvelle génération de patches composés de matériaux cliniques et de couches multiples de bandes synchrones de cardiomyocytes. De fait, ces travaux ouvrent de nouvelles voies dans l’utilisation de biomatériaux pour la production industrielle de cardiomyocytes et pour la fabrication de patches cliniques, cellularisés ou non, dans le traitement de l’insuffisance cardiaqueCell therapy is a promising strategy to help regenerate the damaged heart. Recent studies have placed a lot of hopes in embryonic stem cells and our lab had previously found a way to differentiate them into cardiac progenitors, cells that can only differentiate into cardiomyocyte, endothelial cells or smooth muscle cells. This early commitment decreases their proliferative capabilities, yet maintains their plasticity for better integration inside the host tissue. However, clinical and pre-clinical injection studies did not really meet the expectations. Even though slight improvements in cardiac function were demonstrated, very low cell viability has been observed, as well as a very low retention of the cells inside the myocardium. To address this problem, my PhD projects not only focus on the design of new biomaterials to act as a vehicle for cell delivery and retention in the infarcted area, but also on the design of biomaterials that control the cellular environment during the differentiation of pluripotent stem cells into cardiomyocytes. Going back and forth between the labs and the clinics, we first developed new techniques for the fabrication and the characterization of a cell-laden fibrin patch that is now undergoing phase I clinical trial. From the approved clinical formulation, we then propose new blends of clinical materials that will eventually improve the maturation of the cardiac progenitors once grafted onto the failing heart. In this perspective, we developed an in vitro model to investigate the combined influence of matrix elasticity and topographical confinement on stem cell differentiation into cardiomyocytes. By using microfabrication techniques to pattern pluripotent stem cells on substrates of controlled stiffness, we demonstrate that even using a widely recognized chemical-based protocol to modulate signaling cascades during differentiation, much heterogeneity emerges depending on the cellular physical environment. We thus extracted the main features that led to controlled and reproducible cardiac differentiation and applied it to the fabrication of next generation of multi-layered anisotropic cardiac patches in compliances with clinical requirements. This work opens new routes to high-scale production of cardiomyocytes and the fabrication of cell-laden or cell-free clinical patches
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Hinze, Arnd [Verfasser], Tilmann [Akademischer Betreuer] Grune, Berit [Akademischer Betreuer] Jungnickel e Ingo [Akademischer Betreuer] Bechmann. "Microglia differentiation and cell therapie / Arnd Hinze. Gutachter: Tilmann Grune ; Berit Jungnickel ; Ingo Bechmann". Jena : Thüringer Universitäts- und Landesbibliothek Jena, 2014. http://d-nb.info/1051621399/34.
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Penton, Christopher M., Vasudeo Badarinarayana, Joy Prisco, Elaine Powers, Mark Pincus, Ronald E. Allen e Paul R. August. "Laminin 521 maintains differentiation potential of mouse and human satellite cell-derived myoblasts during long-term culture expansion". BIOMED CENTRAL LTD, 2016. http://hdl.handle.net/10150/622727.
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Background: Large-scale expansion of myogenic progenitors is necessary to support the development of high-throughput cellular assays in vitro and to advance genetic engineering approaches necessary to develop cellular therapies for rare muscle diseases. However, optimization has not been performed in order to maintain the differentiation capacity of myogenic cells undergoing long-term cell culture. Multiple extracellular matrices have been utilized for myogenic cell studies, but it remains unclear how different matrices influence long-term myogenic activity in culture. To address this challenge, we have evaluated multiple extracellular matrices in myogenic studies over long-term expansion. Methods: We evaluated the consequence of propagating mouse and human myogenic stem cell progenitors on various extracellular matrices to determine if they could enhance long-term myogenic potential. For the first time reported, we comprehensively examine the effect of physiologically relevant laminins, laminin 211 and laminin 521, compared to traditionally utilized ECMs (e.g., laminin 111, gelatin, and Matrigel) to assess their capacity to preserve myogenic differentiation potential. Results: Laminin 521 supported increased proliferation in early phases of expansion and was the only substrate facilitating high-level fusion following eight passages in mouse myoblast cell cultures. In human myoblast cell cultures, laminin 521 supported increased proliferation during expansion and superior differentiation with myotube hypertrophy. Counterintuitively however, laminin 211, the native laminin isoform in resting skeletal muscle, resulted in low proliferation and poor differentiation in mouse and human cultures. Matrigel performed excellent in short-term mouse studies but showed high amounts of variability following long-term expansion. Conclusions: These results demonstrate laminin 521 is a superior substrate for both short-term and long-term myogenic cell culture applications compared to other commonly utilized substrates. Since Matrigel cannot be used for clinical applications, we propose that laminin 521 could possibly be employed in the future to provide myoblasts for cellular therapy directed clinical studies.
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Ullah, Mujib [Verfasser]. "Molecular characterization of human mesenchymal stem cell differentiation to identify biomarkers for quality assurance in stem cell therapy / Mujib Ullah". Berlin : Medizinische Fakultät Charité - Universitätsmedizin Berlin, 2014. http://d-nb.info/1047579197/34.
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Espinha, Nuno Miguel Moura. "Bioprocess engineering of induced pluripotent stem cells for application in cell therapy and pre-clinical research". Master's thesis, Faculdade de Ciências e Tecnologia, 2014. http://hdl.handle.net/10362/11551.
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DeSmet, Sara. "Exploring Therapeutic Outcomes Through Picture Books, Other Stories, and Art Therapy". Digital Commons at Loyola Marymount University and Loyola Law School, 2012. https://digitalcommons.lmu.edu/etd/106.
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Therapeutic outcomes are explored in a series of case studies where art therapy and storytelling interventions are used with clients. Stories utilized in the study include picture book stories, fairytales, and self-generated narratives. Additionally, the study’s participants created art responses that took such forms as illustrations and altered books. Research questions that were investigated were: When children receiving art therapy engage with stories created by others or the author, how do they respond?; When children receiving art therapy create their own stories, how do they respond?; and Is there archetypal or other psychologically meaningful content in the author’s picture book? Main subjects of the study were clients ages 9 to 12 receiving individual and group therapy services from the author at The Whole Child in Whittier, California. The author was also a subject in the study. She studied her picture book for significant content. A case study approach was used to highlight themes of psychological or therapeutic relevance for all participants. Biographical data as well as responses to interventions were recorded in assessment and progress notes. Additionally, the therapist shared a piece of her own creative writing for each case study in order to understand clients through the storytelling process. Then the biographical data, story, art responses, and creative writing pieces were studied to look for any connections and to draw conclusions. Based on these results, it appears that sharing pre-existing stories with clients or asking clients to create their own stories has therapeutic value. Not only did these interventions appear to aid clients’ expressions, but they also helped the therapist gain important understandings about clients. Similar analysis of the author’s picture book brought to light themes of psychological importance that increased her self-understanding.
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Holowacz, Eugene holowacz. "Understanding Differentiation of Self Through an Analysis of Individuality and Togetherness". The Ohio State University, 2016. http://rave.ohiolink.edu/etdc/view?acc_num=osu1470938547.
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Deiser, Katrin. "Impact of IL-7 signaling on adoptive T cell therapy". Doctoral thesis, Humboldt-Universität zu Berlin, Lebenswissenschaftliche Fakultät, 2016. http://dx.doi.org/10.18452/17419.
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Das Zytokin Interleukin-7 (IL-7) ist für die Entstehung und das Überleben reifer T Zellen von zentraler Bedeutung. Die Gabe von IL-7 führt sowohl in der Maus als auch im Menschen zu erhöhten T Zellzahlen und einem veränderten T Zellphänotyp. Folglich könnte sich die therapeutische Gabe von IL-7 bei Patienten mit geschwächtem Immunsystem positiv auswirken. Diese Hypothese wird derzeit in mehreren klinischen Studien untersucht. Bisher wurde allerdings nur die Wirkung von IL-7 auf T-Zellen studiert. Zu dessen Wirkung auf andere Immun- oder Stromazellen sowie deren IL-7-abhängigen Beitrag zur Regulation der T-Zellhomöostase ist nur wenig bekannt. Daher war es Ziel der Arbeit, den Einfluss einer therapeutischen Gabe von IL-7 auf adoptiv-transferierte T-Zellen in IL-7-Rezeptor (IL-7R)-kompetenten und defizienten lymphopenischen Mäusen zu studieren. Die Untersuchungen bestätigen, dass die Gabe von IL-7 T-Zellantworten unterstützt, zeigen jedoch auch, daß viele dieser Effekte von IL-7R-exprimierenden Wirtszellen abhängig sind. Dies weist darauf hin, dass IL-7R-vermittelte Signale in Wirtszellen indirekt T-Zellantworten beeinflussen. Zudem zeigte sich, dass effiziente anti-Tumor-T Zellantworten von IL 7R-vermittelten Signalen in Wirtszellen abhängen. Vor allem nicht-hämatopoetische Wirtszellen fungieren hier als Regulatoren der IL-7-Therapie-vermittelten T Zelldifferenzierung. Unsere Ergebnisse bestätigen außerdem, dass Stromazellen in verschiedenen Organen il-7 exprimieren und zeigen darüber hinaus, dass diese Zellen durch die Gabe von IL-7 beeinflusst werden. Wir folgern daraus, dass die Effekte der IL-7-Therapie auf T Zellhomöostase teilweise indirekt über il-7-exprimierende Stromazellen vermittelt werden. Um diese Zellen genauer identifizieren und untersuchen zu können, haben wir ein neues transgenes Mausmodell charakterisiert, was es erleichtern wird, die beteiligten molekularen Signalwege zu analysieren und den Erfolg der adoptiven T Zelltherapie zu verbessern.Interleukin-7 (IL-7) is an essential cytokine required for the development and maintenance of mature T cell. Its availability is limited under normal conditions, but rises during lymphopenia, leading to increased T cell proliferation. The administration of recombinant IL-7 to normal or lymphopenic mice and humans results in increased T cell numbers and altered T cell phenotype. Hence, IL-7 administration could mediate therapeutic benefits in immunocompromised patients and is currently tested in several clinical trials. However, besides its well-studied effects on T cells little is known about the effect of IL-7 on other immune and non-immune cells and their influence on T cell homeostasis. Therefore, we evaluated the effect of IL-7 therapy on adoptively transferred T cells in IL-7 receptor (IL-7R)-competent and IL-7R-deficient lymphopenic mice. We confirm the benefits of IL-7 therapy on T cell responses but additionally show that many of these effects are dependent on IL-7R expression by host cells, indicating that IL-7R signaling in host cells modulates T cell responses. We show that efficient T cell responses against cancer are dependent on host IL-7R signaling. Based on studies in bone-marrow chimeric mice, we identify non-hematopoietic host cells as main regulators of IL-7 therapy-modulated T cell differentiation. We conclude from these data that IL-7 therapy affects non-hematopoietic stromal cells that modulate the success of adoptive T cell therapy. Our results confirm that stromal cells in various organs express il-7 and show that these cells are targeted by IL-7 therapy in vivo. Hence, we propose that il-7-expressing cells regulate IL-7 therapy-modulated T cell homeostasis. To identify and study these il-7 expressing stromal cells in more detail, we characterized a new transgenic mouse model that will facilitate determining the molecular pathways to improve the success of adoptive T cell therapy.
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Smidova, Eva. "Transgender Parent Differentiation: A Heuristic Phenomenological Study". Diss., NSUWorks, 2019. https://nsuworks.nova.edu/shss_dft_etd/50.
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Transgender individuals go through their intrapersonal differentiation between covert “I” (expressed gender) and overt “I” (assigned gender), often unnoticed by family members before their coming out. Consequently, their coming out rockets anxiety in the family system and the process of differentiation of transgender parents seem to go through its unique path to search for equilibrium. Recent social and clinical studies about transgender parents have paid attention to the experience and challenges of the gender transition process, social pressure, acceptance of transgender individuals in a parenting role, and readiness of families to cope with the transition of a parent (Bischof, Warnaar, Barajas, & Dhaliwal, 2011; Chung, 2016; Di Ceglie, 1998; Freedman, Tasker, & Di Ceglie, 2002; Haines, Ajayi, & Boyd, 2014; Hines, 2006; Theron & Collier, 2013; Veldorale-Griffin, 2014; White & Ettner, 2004, 2007). No research study has attempted to explore the essence of transgender parenting and the related self-differentiation process (Bowen, 1978; Kerr & Bowen, 1988). In this research, I intended to address this gap in knowledge by utilizing a heuristic phenomenological research design to explore the essence of parenting and self-differentiation of transgender parents. I used interviews with ten transgender parents, both females, and males, to embrace the elements of the lived experienced. The first conducted heuristic analysis revealed five emerging themes: Selfish Unselfishness: Becoming Me; Relationship with My Close Family: It is About Respect; Battle of Emotions: Do the Right Thing; Competence, Confidence, and Legacy: This Is How We Do It. Or Not; and Life Satisfaction: Welcome to My World. The second, qualitative data analysis, brought evidence of these qualities of self-differentiation: Balancing Individuality and Togetherness; Balancing Thoughts and Feelings; and Self-differentiation in the Expressed Gender.
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Li, Xiaofei. "Improving Stem Cell Survival and Differentiation in Ischemic and Inflammatory Tissues". The Ohio State University, 2016. http://rave.ohiolink.edu/etdc/view?acc_num=osu1469174124.
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Glade, Aaron C. "Differentiation, marital satisfaction, and depressive symptoms: an application of Bowen Theory". The Ohio State University, 2005. http://rave.ohiolink.edu/etdc/view?acc_num=osu1123257865.
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MATHOR, MONICA B. "Estudos da expressao genica mediante utilizacao de queratinocitos humanos normais transduzidos com o gene do hormonio de crescimento humano .Possivel utilizacao em terapia genica". reponame:Repositório Institucional do IPEN, 1994. http://repositorio.ipen.br:8080/xmlui/handle/123456789/10410.
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Made available in DSpace on 2014-10-09T12:38:22Z (GMT). No. of bitstreams: 0Made available in DSpace on 2014-10-09T14:05:27Z (GMT). No. of bitstreams: 1 05680.pdf: 8681194 bytes, checksum: c738dd10c1a17a2b018e74273b788729 (MD5)
Tese (Doutoramento)
IPEN/T
Instituto de Pesquisas Energeticas e Nucleares - IPEN/CNEN-SP
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Yoo, Hana. "Couple Intimacy and Relationship Satisfaction: A Comparison Study between Clinical and Community Couples". The Ohio State University, 2013. http://rave.ohiolink.edu/etdc/view?acc_num=osu1374180064.
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Mekhail, Mina. "Injectable purine-based sponges to promote oligodendrocyte progenitor cells' attachment and differentiation: a potential therapy to target remyelination post-spinal cord injuries". Thesis, McGill University, 2014. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=123217.
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Demyelination is one of the adverse events that persist during both the acute and chronic stages of spinal cord injury (SCI). Intact axons that survive the initial trauma but lose their myelin undergo degeneration if myelin is not re-instated. Incomplete remyelination takes place by an endogenous population of progenitor cells called Oligodendrocyte Progenitor Cells (OPCs). The consensus in the literature is that abnormal myelin is a result of the inability of OPCs to undergo complete differentiation at the site of injury. Therefore, providing an environment conducive of OPC differentiation after a SCI can improve the remyelination process and restore some neurological functions. Biomaterials have been widely developed for promoting neural regeneration post-SCI; however, there are very few reports on biomaterials developed specifically to enhance remyelination post-SCI. Therefore, the main overarching objective of this thesis was to develop a biomaterial-based approach to enhance OPCs' differentiation. A novel, injectable, rapidly-gelling, Guanosine 5'-diphosphate (GDP)-crosslinked chitosan sponge was developed to specifically target OPC survival and differentiation. GDP has never been investigated as an anionic crosslinker, and was explored in this thesis because it contains two key components that serve two functions simultaneously. The phosphate groups ionically crosslink the amine groups in chitosan and induce rapid gelation (tgel< 1.6 seconds). On the other hand, previous studies have shown that guanosine improved remyelination in situ post-SCI, and therefore its presence in the chitosan sponge composition was hypothesized to promote OPC survival, attachment and differentiation. The physicochemical properties of the chitosan sponges were characterized and their cytocompatibility was assessed in vitro. The sponges are highly porous, can retain water up to 10 times their own weight, have moduli of elasticity close to that of the spinal cord, and are cytocompatible with a variety of cell types including OPCs. These chitosan sponges are also able to encapsulate growth factors (NT-3 and BDNF) with high efficiencies (> 70%) and provide a controlled release for more than 2 weeks. Rat-derived OPCs adhere on the sponges and acquire physiological phenotypes. Moreover, differentiation on the sponges is more significant than controls even in the presence of mitogens. OPCs cultured on the sponges for 8 days express myelin basic protein (MBP), a marker for mature oligodendrocytes. Human fetal OPCs also adhere and differentiate on the sponges. Finally, a double-barrel, double-lumen injection system has been designed to administer the chitosan sponge in situ and was tested successfully. Our findings suggest that the injectable chitosan sponges presented in this thesis are a clinically-relevant therapeutic modality for improving remyelination post-SCI.La démyélinisation est une conséquence perpétuelle des phases aigües et chroniques d'une lésion de la moelle épinière (ou LME). La perte de myéline entourant les axones intacts est inévitable si la remyélinisation n'a pas lieu. Cette remyélinisation est suscitée par une population endogène de cellules progénitrices appelées précurseurs d'oligodendrocyte (ou CPO). Cependant, ce processus de remyélinisation est incomplet et engendre la formation anormale de la gaine de myéline. Selon la littérature scientifique existante, cette myéline anormale est due à l'incapacité de différenciation des OPC dans la zone atteinte. Il faut donc fournir aux OPC un environnement adapté qui peut induire leur différenciation à la suite d'une LME pour ainsi améliorer le processus de remyélinisation et rétablir certaines fonctions neurologiques.Bien que l'étude de biomatériaux dans la régénération neuronale laisse entrevoir des résultats prometteurs, il n'y a présentement pas de publication sur les biomatériaux ciblant précisément la démyélinisation. L'objectif principal de cette thèse est donc de développer un biomatériau promouvant la survie et la différenciation des OPC. Une nouvelle éponge injectable et à gélification rapide a été mise au point en effectuant la réticulation de chitosane et de la guanosine 5'- diphosphate (GDP). La GDP, qui n'a jamais été étudiée comme agent de réticulation anionique auparavant, a été analysée pour ses deux éléments clés visant à remplir deux fonctions combinées. Les groupes de phosphate réticulent ioniquement les groupes aminés se trouvant dans le chitosane permettant ainsi une gélification rapide (Tgel < 1,6 secondes). D'autre part, des études antérieures ont démontré que la GDP améliore la remyélinisation in situ suivant une LME. On a donc émis l'hypothèse que la présence GDP dans cette éponge de chitosane servirait à promouvoir la survie, l'adhésion ainsi que la différenciation des OPC.Les propriétés physico-chimiques de cette éponge à base de chitosane ont été caractérisées et sa biocompatibilité a été évaluée in vitro. L'éponge s'est révélée être d'une grande porosité, pouvant retenir jusqu'à dix fois son poids en eau. De plus, le module d'élasticité de l'éponge se rapproche de celle de la moelle épinière. Des analyses ont démontré que l'éponge est cytocompatible avec toute une variété de lignées cellulaires, y compris les OPC. Cette éponge de chitosane a également été utilisée pour encapsuler des facteurs de croissance tel que NT-3 et BDNF, démontrant de hauts rendements d'encapsulation (> 70%) et un profil de libération contrôlée sur une période allant au-delà de 14 jours. Des OPC provenant de rats ont été cultivées sur des éponges de chitosane; elles ont démontré une adhérence aux éponges et ont éventuellement acquis des phénotypes physiologiques favorables. De plus, la différenciation des OPC était plus significative sur les éponges que sur les témoins, même en présence de mitogènes suppresseurs de différenciation. Des OPC cultivées sur les éponges pendant 8 jours ont exprimées MBP, un marqueur d'oligodendrocytes matures. Des OPC fœtales humaines cultivés sur les éponges ont également démontré une adhésion et différenciation cellulaire. Enfin, un système d'injection double muni de deux canaux séparés a été conçu pour administrer l'éponge in situ, et a été testé avec succès. Dans l'ensemble, nos résultats démontrent que l'éponge de chitosane injectable présentée dans cette thèse est un modèle de thérapie cliniquement pertinent pour améliorer la remyélinisation suite à une LME.
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Martinez, Ruben G. "Sticking to the recipe: How do adherence and differentiation to a CBT protocol affect client outcomes in youths with anxiety?" VCU Scholars Compass, 2017. http://scholarscompass.vcu.edu/etd/4728.
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Objective: Understanding the pathways through which treatments work to change symptom and diagnostic outcomes is important to the development and delivery of evidence-based treatments. This study assessed the extent to which adherence (therapist’s delivery of prescribed therapeutic interventions) and differentiation (therapist’s delivery of non-prescribed therapeutic interventions) to Coping Cat, a CBT program, affected client symptom and diagnostic outcomes. Method: The Therapy Process Observational Coding System for Child Psychotherapy – Revised Strategies Scale (McLeod et al., 2015) was used to characterize therapeutic interventions delivered within and outside of the Coping Cat program with youths aged 7-15 receiving treatment in one efficacy (n = 51; 41% female; 84% Caucasian, M age = 10.37) and one effectiveness (n = 17; 56% female, 39% Caucasian, M age = 10.90) trial. Youth- and parent-report symptom checklists and diagnostic interviews were used to assess symptom and diagnostic remission. Multiple hierarchical regression analyses and hierarchical binomial logistic regression were used to investigate the relation between adherence and differentiation and symptom change and remission of principal diagnosis. Results: Neither adherence nor differentiation were significantly related to symptom or diagnostic outcomes. No clear trend emerged, and results were inconsistent across parent and youth report, outcome type, and setting. Conclusion: These results are consistent with past literature. Two interpretations exist: (1) that there is no relation between treatment delivery and outcomes, and (2) that methodological and analytic flaws undercut the ability of the analyses to identify a relation.
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Tincho, Marius Belmondo. "In-silico optimization and molecular validation of putative anti-HIV antimicrobial peptides for therapeutic purpose". University of the Western cape, 2016. http://hdl.handle.net/11394/5656.
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Philosophiae Doctor - PhDAIDS is considered a pandemic causing millions of deaths worldwide and a cure for this disease is still not available. Failure to implement early treatments due to the poor diagnostic methods and ineffective therapeutic regimens to treat HIV patients to achieve complete viral eradication from the human body has encouraged the escalation of this disease at an exponential rate. Though the current treatment regimens (High Active Antiretroviral Therapy) have aided in increasing the lifespan of HIV patients, it still suffers from some shortcomings such as adverse side effects and non-eradication of the virus. Thus, there is a need for a non-toxic therapeutic regimen to stop further infection of HIV-infected patients. Antimicrobial Peptides (AMPs) are naturally occurring peptides which are components of the first line of defence of many organisms against infections and have been proven to be promising therapeutic agents against HIV. The use of AMPs as anti-microbial agents is due to the fact that most AMPs have a net positive charge and are mostly hydrophobic molecules. These features allow AMPs to be site directed electro-statistically to the mostly negatively charged pathogens. In a previous study, a number of novel anti-HIV AMPs was identified using a predictive algorithm Profile Hidden Markov Models (HMMER). The AMP's threedimensional structures were predicted using an in-silico modelling tool I-TASSER and an insilico protein-peptide interaction study of the AMPs to HIV protein gp120 was performed using PatchDock. Five AMPs were identified to bind gp120, at the site where gp120 interacts with CD4 to prevent HIV invasion and HIV replication. Therefore, the aims of this research were to perform in-silico site-directed mutation on the parental anti-HIV AMPs to increase their binding affinity to the gp120 protein, validate the anti-HIV activity of these peptides and confirm the exclusivity of this activity by testing possible anti-bacterial and anti-cancer activities of the AMPs. Firstly, the five parental anti-HIV AMPs were used to generate mutated AMPs through insilico site-directed mutagenesis. The AMPs 3-D structures were determined using I-TASSER and the modelled AMPs were docked against the HIV protein gp120 using PatchDock. Secondly, an "in house" Lateral Flow Device (LFD) tool developed by our industrial partner, Medical Diagnostech (Pty) Ltd, was utilised to confirm the in-silico docking results. Furthermore, the ability of these AMPs to inhibit HIV-1 replication was demonstrated and additional biological activities of the peptides were shown on bacteria and cancer cell lines. In an effort to identify AMPs with increased binding affinity, the in-silico results showed that two mutated AMPs Molecule 1.1 and Molecule 8.1 bind gp120 with high affinity, at the point where gp120 bind with CD4. The molecular binding however showed that only Molecule 3 and Molecule 7 could prevent the interaction of gp120 protein and CD4 surface protein of human cells, in a competitive binding assay. Additionally, the testing of the anti-HIV activity of the AMPs showed that Molecule 7, Molecule 8 and Molecule 8.1 could inhibit HIV-1 NL4-3 with maximal effective concentration (EC₅₀) values of 37.5 μg/ml and 93.75 μg/ml respectively. The EC₅₀ of Molecule 8.1 was determined to be around 12.5 μg/ml. This result looks promising since 150 μg/ml of the AMPs could not achieve 80% toxicity of the human T cells, thus high Therapeutics Index (TI) might be obtained if 50% cytotoxic concentration (CC₅₀) is established. Further biological activity demonstrates that Molecule 3 and Molecule 7 inhibited P. aeruginosa completely after 24 hours treatment with peptide concentrations ranging from 0.5 mg/ml to 0.03125 mg/ml. Nevertheless, moderate inhibition was observed when CHO, HeLa, MCF-7 and HT-29 were treated with these peptides at peptides concentration of 100 μg/ml. The ability of these AMPs to block the entrance of HIV via the binding to CD4 of the host cells is a good concept since they pave the way for the design of anti-HIV peptide-based drugs Entry Inhibitors (FIs) or can be exploited in the production microbicide gels/films to suppress the propagation of the virus.
DST-NIC/Mintek
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Waddoups, Samuel Dean. "Evaluation of critical parameters of low level laser irradiation on human osteoblast cell proliferation and differentiation". Thesis, NSUWorks, 2012. https://nsuworks.nova.edu/hpd_cdm_stuetd/60.
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Abstract
Orthodontic tooth movement is a biological response to a mechanical force. One of the challenges in orthodontics is obtaining desired tooth movement during treatment. Accelerating tooth movement and decreasing demands on anchorage can reduce treatment times and overall satisfaction for patient and doctor. Low-level laser therapy (LLLT) is emerging as a technology that may decrease orthodontic treatment time. Many in vitro and in vivo studies have reported the effects of low level lasers at random time points and energy densities. None of the studies have optimized the dose required for osteoblast proliferation and differentiation. The purpose of this study was to find the optimum stimulatory dose of low level laser irradiation (LLLI) on human osteoblast cell proliferation and differentiation and to analyze our findings with reference to the Arndt-Shultz Law of applied energy. In this in vitro study a GaAlAs laser at 830nm, 20 mW with continuous exposure at various doses were used on a human osteoblast cell line. According to the Arndt-Shulz Law weak stimuli initiate vital activity, moderate stimuli enhance the cellular activity with subsequent peak stimulation and greater stimuli (beyond a threshold value) may not have any influence or inhibit the vital activity. The implications of LLLI on human osteoblasts and influencing tooth movement in orthodontics were discussed. Human osteoblasts were cultured in minimum essential medium (MEM) complete medium consisting 10% fetal bovine serum and 1% antibiotics. Cells grown in complete medium were plated onto 96 well plate, allowed to adhere for 4-5 hours and were exposed to GaAlAs lasers at 6 , 12, 18, 24, 30, 36, 45, 60, 75, and 90 seconds. The cells treated with xiii LLLI were assessed for cell proliferation at 24, 48 and 72 hour intervals. A calorimetric cell proliferation assay (WST-1) assay was performed according to manufacture's instructions. The results indicated that at 24 hours the 6 and 12 seconds doses significantly inhibited proliferation compared to the control. At 48 hours the 30 seconds exposure significantly increased proliferation. At 72 hours time interval, cell proliferation was observed in a dose dependent pattern with a minimum at 6 seconds with peak proliferation at 18 seconds. A gradual decrease in cell viability was observed in the cells treated beyond this dose with a maximum inhibition seen at 60 seconds. At 75 and 90 seconds no difference was observed between the control and experimental group. To establish efficient acquisition of adequate quantities of alkaline phosphatase, cells were grown in 12 well plates in complete medium or osteogenic medium. These cells were exposed to LLLI for 18, 48, and 60 seconds. The activity of early osteogenic differentiation marker alkaline phosphatase (ALP) was investigated 10 days post exposure. Our results demonstrated that alkaline phosphatase activity at 2.4 - 7.3 J/cm2 with 48 - 60 seconds of exposure, and an incident power ranging from 85-269mw significantly increased. The findings suggest that these irradiated cells obeyed the Arndt Shulz Law governing cellular response to applied energy. Further this research indicates the possible role of LLLT to accelerate tooth movement in orthodontics. Complete disclosure of low level laser parameters is essential in order to accurately compare findings of researchers.
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Bischof, Ashley Gibbs. "Extracellular Matrix as a Key Mediator of Mammary Tumor Cell Normalization". Thesis, Harvard University, 2013. http://dissertations.umi.com/gsas.harvard:10780.
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Some epithelial cancers can be induced to revert to quiescent differentiated tissues when combined with embryonic mesenchyme; however, the mechanism of this induction is unknown. This dissertation is based on the hypothesis that because extracellular matrix (ECM) plays a critical role during organ development in the embryo, it also may mediate the differentiation-inducing effects of embryonic mesenchyme on cancer cells. To test this hypothesis, I first optimized methods to isolate ECMs from whole tissues or cultured cells, and to repopulate them with cultured cells, using embryonic tooth as a model system. In Chapter 2, I describe these studies and use them to demonstrate that embryonic ECM is sufficient to regulate odontogenic signaling, cell fate decisions and histodifferentiation during normal tooth development. In Chapter 3, I adapt these methods to show that culture of breast cancer cells with ECM derived from embryonic mammary mesenchyme decreases tumor cell proliferation, and stimulates differentiation, including formation of hollow acini and ducts as well as enhanced expression of estrogen receptor-alpha and decreased migration. Further, when the inductive ECMs were injected into fast-growing breast tumors in mice, they significantly inhibited cancer expansion. Critically, the differentiation observed with ECM was the same as that observed in co-culture with mammary mesenchyme cells, showing that ECM is playing a dominant role in tumor cell normalization. In Chapter 4, I then set out to determine the mechanism by which embryonic ECM normalizes tumor cells, I analyzed the contributions of bound cytokines, ECM composition and mechanics. Western blot analysis revealed several bound growth factors, which remained following decellularization; however, removal of these growth factors using high salt washes had no effect on ECM-mediated normalization of tumors. Further, using proteomics analysis I identified eleven ECM proteins present only within inductive ECMs and by testing these proteins in 3D culture, I found three proteins -- collagen III, biglycan and SPARC -- that increased lumen formation to a similar extent as embryonic ECM. These data confirm that mesenchyme-induced tumor cell normalization is mediated by the insoluble ECM, and reveal the identity of some of the inductive molecules responsible for these effects.
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Rollakanti, Kishore Reddy. "Protoporphyrin IX Fluorescence for Enhanced Photodynamic Diagnosis and Photodynamic Therapy in Murine Models of Skin and Breast Cancer". Cleveland State University / OhioLINK, 2015. http://rave.ohiolink.edu/etdc/view?acc_num=csu1431466604.
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Cochonneau, Stéphanie. "Modulating hematopoietic progenitor cell engraftment and T cell differentiation : role of conditioning and route of administration". Thesis, Montpellier 2, 2012. http://www.theses.fr/2012MON20226.
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Les déficits lymphocytaires T peuvent être corrigés par l'administration en intraveineuse (IV) de cellules souches hématopoiétiques (CSH) provenant d'un donneur. Dans un modèle d'immunodéficience lié à l'absence de la protéine kinase ZAP-70, notre équipe avait précédemment montré que l'injection intrathymique (IT) de CSH histocompatibles conduit à une reconstitution du compartiment T plus robuste et plus rapide que dans le cas où les CSH sont administrées par voie IV. Au cours de ma thèse, je me suis intéressée à l'approche IT dans un contexte non-histocompatible, où j'ai montré que l'injection de CSH semi-allogéniques directement dans le thymus permet le développement d'une thymopoièse à long-terme, même en absence de conditionnement. De plus, j'ai également montré la persistence de progéniteurs thymiques précoces (ETP) provenant du donneur dans le thymus des souris transplantées. De façon remarquable, ces ETP retiennent un potentiel de différenciation plus divers que ceux rencontrés dans le thymus d'une souris sauvage, et leur fréquence est significativement élévée après IT, ce dernier suggérant une disponibilité accrue des niches thymiques. De façon intéressante, j'ai également montré que les progéniteurs déficients en ZAP-70 pouvaient se différencier de façon importante vers le lignage CD8 lors d'une activation constante de la voie de signalisation Notch couplée à la présence d'interleukine 7 (IL-7). Après la greffe de CSH par voie IV de souris ZAP-70-/-, en absence de conditionnemt, j'ai également identifié l'accumulation d'une population de CSH présentant un phénotype particulier (Lin- Sca 1+ c-kit-), nommée LSAPT. Ces cellules LSAPT présentent un biais de différenciation vers le lignage T γδ ainsi qu'une production élevée d'IL-17, ce qui suggère que les fonctions effectrices d'une cellule T γδ sont dépendantes de leur origine progénitrices. L'ensemble de mes résultats apporte à la fois de nouveaux éléments concernant l'identification de progéniteurs T et démontrent de l'influence/coopération entre voies de signalisation et facteurs environnementaux dans la modulation de la différenciation T et de leur fonctions effectricesT cell deficiencies can be corrected by the intravenous (IV) injection of donor hematopoietic stem cells (HSCs). Using a murine model of ZAP-70-/- deficiency, our group previously showed that the intrathymic (IT) administration of histocompatible HSCs leads to a more robust and long-term thymopoiesis as compared to that achieved by the classical IV route. During my PhD, I found that the direct IT administration of semiallogeneic HSCs results in a sustained donor-derived thymopoiesis, overcoming histocompatibility barriers, even in the absence of conditioning. Furthermore, I found that donor-derived early thymic progenitors (ETPs) persist in the thymi of ZAP-70-/- transplanted mice, and present increased multi-lineage potential as compared to wild-type ETPs. Importantly, the frequency of donor-derived ETPs was augmented following IT transplantation, indicative of an increased progenitor niche. Interestingly, ZAP-70-deficient HSC could themselves be driven to a CD8 lineage fate in an environment where IL-7 potentiates continuous activation of the Notch pathway. Following IV transplantation of donor HSC into non-conditioned ZAP-70-/- mice, I determined that there is an accumulation of lineage-/Sca1+ donor progenitors lacking expression of the stem cell marker c-kit, termed LSAPT. These LSAPT show a biased differentiation towards the γδ T cell lineage with high IL-17-producing effector function, suggesting that progenitor origin regulates γδ T cell fate. The ensemble of my experiments provide new insights into the identity of T lineage progenitors and demonstrate how signaling pathways as well as environmental factors modulate T cell differentiation and effector function
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Tsuji, Hideaki. "Multicenter Prospective Study of the Efficacy and Safety of Combined Immunosuppressive Therapy With High-Dose Glucocorticoid, Tacrolimus, and Cyclophosphamide in Interstitial Lung Diseases Accompanied by Anti-Melanoma Differentiation-Associated Gene 5-Positive Dermatomyositis". Kyoto University, 2021. http://hdl.handle.net/2433/261610.
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Kim, Sun Wook. "Modulation of Stem Cell Fate by Electrical Stimulation". University of Cincinnati / OhioLINK, 2013. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1383812480.
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Mercado, Alexis. "Addiction and the Family: Substance Use as a Symptom of the Larger Emotional System". Diss., NSUWorks, 2019. https://nsuworks.nova.edu/shss_dft_etd/49.
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Traditional family therapy in the field of addiction primarily focuses on relapse prevention and psychoeducation. The lack of systems thinking in residential treatment facilities led to my desire to apply Bowen Family Therapy to a focus group in a residential treatment center. I used the following Bowen concepts: anxiety, differentiation of self, emotional cutoffs, and triangulation as a means to explore how addiction is a symptom of the larger emotional system of the family. I, co-facilitated a three hour group therapy session over 7 weeks with individuals in a treatment center. I addressed the following questions: RQ 1: What impact, if any did this program have on their life? RQ 2: What were the long-term effects of being in the program? RQ 3: Did participating in the group help to better understand resiliency? RQ 4: How does education on the family system impact an individual's recovery process and relationships in life? Through interviews, I followed up with clients three years later to look at the long-term effects of being in the 7-week program. This Applied Clinical Project focused on understanding resiliency and long-term effects on sobriety through a Bowenian lens. The themes that emerged focused on communication, boundaries, resiliency, relationships, and anxiety. The findings demonstrated that a multigenerational element in the study helped participants develop a way to maintain the Family Dynamics curriculum in their day to day life. The overarching theme is that healthy relationships with open communication lead to better anxiety management, resiliency, and boundaries which shows a foundation of which new approaches to substance abuse treatment can be found.
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Cully, Laura Marie. "Predictors of Intimate Partner Violence among Women Seeking Treatment for a Substance Use Disorder". The Ohio State University, 2018. http://rave.ohiolink.edu/etdc/view?acc_num=osu152267976367848.
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Cleland, Nicole Rae Cleland. "Differentiation of Self and Effortful Control: Predictors of Non-Traditional Students' Adjustment to Community College". University of Akron / OhioLINK, 2017. http://rave.ohiolink.edu/etdc/view?acc_num=akron1509913708613883.
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Palmer, Elizabeth Northup Palmer. "Using distance regulation for the study of sibling relationship quality, romantic relationships, and interpersonal and intrapersonal factors". The Ohio State University, 2017. http://rave.ohiolink.edu/etdc/view?acc_num=osu1500469586490535.
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Montacir, Houda [Verfasser]. "Glycan-based biomarkers for the quality assurance in stem cell therapy : Cell surface N-glycosylation of human stem cells and its changes during the process of differentiation / Houda Montacir". Berlin : Freie Universität Berlin, 2014. http://d-nb.info/1053959605/34.
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Ignatz-Hoover, James J. "TLR8 and Nuclear GSK3ß are Novel Therapeutic Targets in AML". Case Western Reserve University School of Graduate Studies / OhioLINK, 2017. http://rave.ohiolink.edu/etdc/view?acc_num=case1480509044211523.
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Zhou, Yinghong. "Interactions between undifferentiated and osteogenically differentiated mesenchymal stromal cells during osteogenesis". Thesis, Queensland University of Technology, 2013. https://eprints.qut.edu.au/63002/1/Yinghong_Zhou_Thesis.pdf.
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The body of work presented in this dissertation has demonstrated that the interactions between donor cells and host cells are critical for bone repair and regeneration. The donor cells secrete VEGF which activates the downstream PI3K/Akt signaling pathway, ultimately leading to host cell recruitment and robust bone regeneration. The findings from this dissertation may provide a scientific rationale for the development of novel therapeutic strategies in the treatment and management of bone defects.
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Caxaria, Sara. "Induced pluripotent stem cells (iPSCs) for research and therapy : induction of hepatic differentiation in iPSCs and evaluation of their quality as a model of in vivo development in the context of coagulation". Thesis, University College London (University of London), 2016. http://discovery.ucl.ac.uk/1474575/.
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Stem cells hold great promise for regenerative medicine as they have the potential to repair almost any tissue. The use of induced pluripotent stem cells (iPSC) offers several advantages over human embryonic stem cells (hESC). Nevertheless, an issue that has slowed the use of iPSC (as well as hESC) in the clinic is safety. The pluripotent capacity that gives these cells their regenerative potential also gives them tumorigenic abilities. Moreover, the reprogramming procedure of iPSC can also affect the quality and safety of the final population. In the search for safer iPSC, we optimized an integration free method of reprogramming that is GMP compliant, which represents a step closer to their clinical use. Hepatocyte-like cells derived from iPSC have proven useful in research, as in disease modelling and toxicity screening, and in the context of cellular therapy could represent a breakthrough for the treatment of liver disorders. We worked on an optimized method that allows quick and efficient hepatocyte differentiation from iPSC. We used this as a model to tap into embryonic development in the context of coagulation, and into the regulators of coagulation, with the goal to better understand coagulation. Similar studies can then be used to study other pathways besides coagulation and help increase the knowledge on the liver and its many functions.
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Prudon, Nicolas. "Integrative study, from the cell to the animal model, of the development of a cell therapy for Parkinson's disease". Electronic Thesis or Diss., Bordeaux, 2024. http://www.theses.fr/2024BORD0071.
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Un ensemble d'études précliniques soutient désormais le développement de thérapies de remplacement cellulaire dérivées de cellules souches pluripotentes pour soulager les symptômes moteurs chez les patients parkinsoniens. Le remplacement de la principale population cellulaire dysfonctionnelle au sein de la maladie, les neurones dopaminergiques A9, est l'objectif principal de ces thérapies. Pour y parvenir, la plupart des approches thérapeutiques impliquent la greffe de suspensions cellulaires de progéniteurs dopaminergiques. Cependant, une quantité considérable de cellules meurent pendant le processus de transplantation, les cellules étant confrontées à l'anoïkis. Une potentielle solution consiste à greffer des préparations solides, c'est-à-dire adopter un format cellulaire 3D. La cryopréservation de ce format reste un obstacle majeur et n'est pas exempte de causer des retards dans le délai avant l'apparition des effets, comme observé avec l'utilisation de suspensions unicellulaires de progéniteurs dopaminergiques cryopréservés. Le travail de cette thèse se concentre sur le développement de microtissus neuraux 3D en tant que thérapie cellulaire pour la maladie de Parkinson. L'utilisation d'une technologie d'encapsulation cellulaire à haut débit associée à des bioréacteurs pour fournir un environnement de culture 3D a permis la différenciation dirigée des hiPSC en microtissus neuraux. La différenciation correcte des microtissus neuraux vers une identité mésencéphalique a été confirmée à l'aide de méthodes orthogonales, en utilisant notamment la qRT-PCR, le RNAseq, la cytométrie en flux et la microscopie fluorescente. L'efficacité des microtissus neuraux a été démontrée de manière dose-dépendante dans des études précliniques, en utilisant le modèle de rat hémiparkinsonien lésé par la 6-OHDA. Les greffes ont été caractérisées par une analyse histologique post-mortem, démontrant la présence de neurones dopaminergiques humains projetant dans le striatum hôte. Le travail présenté ici est la première bioproduction d'une thérapie cellulaire pour la maladie de Parkinson dans un bioréacteur mis à échelle, conduisant à une récupération fonctionnelle complète des animaux 16 semaines après la transplantation d’un format cellulaire 3D cryopréservéA breadth of preclinical studies is now supporting the rationale of pluripotent stem cell-derived cell replacement therapies to alleviate motor symptoms in Parkinsonian patients. Replacement of the primary dysfunctional cell population in the disease, i.e. the A9 dopaminergic neurons, is the major focus of these therapies. To achieve this, most therapeutical approaches involve grafting single-cell suspensions of DA progenitors. However, a considerable number of cells die during the transplantation process, as cells face anoïkis. One potential solution to address this challenge is to graft solid preparations, i.e. adopting a 3D format. Cryopreserving such format remains a major hurdle and is not exempt from causing delays in the time to effect, as observed with the use of cryopreserved single-cell DA progenitors. The work of this thesis focus on the development of 3D neural microtissues as a cell therapy for PD. The use of a high-throughput cell-encapsulation technology coupled with bioreactors to provide a 3D culture environment enabled the directed differentiation of hiPSCs into neural microtissues. The proper patterning of these neural microtissues into a midbrain identity was confirmed using orthogonal methods including qPCR, RNAseq, flow cytometry and immunofluorescent microscopy. The efficacy of the neural microtissues was demonstrated in a dose-dependent manner in non-clinical studies, using the 6-OHDA-lesioned hemiparkinsonian rat model. The grafts were characterized by post-mortem histological analysis, demonstrating the presence of human dopaminergic neurons projecting into the host striatum. The work reported here is the first bioproduction of a cell therapy for Parkinson’s disease in a scalable bioreactor, leading to a full behavioural recovery 16 weeks in the animal model after transplantation using cryopreserved 3D cell format