Dissertations / Theses: 'MEL cells'

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Clements, Andrew R. N. "The regulation of globin gene expression." Thesis, University of Oxford, 2000. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.365687.

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Moka, Nagaishwarya, Kelley cross, Marianne Brannon, Janet Lightner, Megan Dycus, William Stone, Victoria Palau, and Koyamangalath Krishnan. "Delta-tocotrienol and simvastatin induces differential cytotoxicity and synergy in BRAF wild-type SK-MEL-2 and mutant BRAF SK-MEL-28 melanoma cancer cells." Digital Commons @ East Tennessee State University, 2018. https://dc.etsu.edu/asrf/2018/schedule/215.

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Targeting the mutant BRAF and immunotherapy are new approaches to the treatment of metastatic malignant melanoma that has significantly improved survival but is associated with significant toxicity and cost. Potent and specific BRAF inhibitors like vemurafenib and dabrafenib are superior to chemotherapy in treatment of BRAF mutant melanomas which represent nearly 50% of all melanomas. A less toxic approach to treatment of malignant melanoma is hence appealing. Delta-tocotrienol (DT3), an unsaturated vitamin E isoform, and simvastatin, an HMG-CoA reductase inhibitor have been shown to have anti-neoplastic properties. We studied the effects of these chemicals in both BRAF-mutated SK-MEL-28 and BRAF-wild type SK-MEL-2 melanoma cells. MTS assays were used to analyze cytotoxicity. SK-MEL-28 and SK-MEL-2 cells were cultured in MEM media containing 10% serum and plated in 96-well culture plates for 48 hours then treated with DT3 (0-80 µM), simvastatin (0-10 µM), or a combination and dosed again at 72 hours. SK-MEL-28 and SK-MEL-2 cells were grown in 60 mm plates and treated with DT3 at concentrations of 30 µM, simvastatin at concentrations of 10 µM and combination of DT3 and simvastatin at concentrations of 10 µM and 2 µM. Cell were lysed with RIPPA buffer with protease and phosphatase inhibitor after 6 hours of treatment. Protein concentration of cell lysates was measured spectrophotometrically (GLO Max Multi+, Promega), using a BCA protein assay kit. The samples were run in SDS PAGE and blotted onto nitrocellulose membranes. Membranes were incubated with antibodies against Hsp 70 (Enzo Life Sciences, Farmingdale, NY), Hsp 90 (Santa Cruz, Dallas, TX), pS6 and pERK (Cell Signaling, Danvers, MA) and pAKT. Using MTS assay, we found that DT3 (IC50 75.2 μM) and simvastatin (IC50 8.3μM) have cytotoxic effects on melanoma cell line SK-MEL-2, but not on the SK-MEL-28 cells DT3 and simvastatin at the concentrations studied (10-80 μM DT3) and (0.625- 10 μM simvastatin). Further studies determined that simvastatin decreased expression of pS6, pERK on SK-MEL-2 and not DT3. However, these effects are different in SK-MEL-28 cells where there is only decrease in expression of pS6; treated SK-MEL-2 cells also show over-expression of Hsp70 suggestive of a rescue effect leading to lesser cytotoxic activity. The selective cytotoxicity observed in wild type BRAF melanoma cell lines by DT3 and simvastatin warrants further research into the potential therapeutic use of these drugs. A differential cytotoxicity is shown by DT3 and simvastatin in malignant melanoma cells with selective more potency in wild type BRAF melanoma compared to mutant BRAF melanoma cells. Further studies will be undertaken to dissect the mechanistic basis of this differential response.

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Garcia-Alonso, Monica. "Evaluation of the potential of murine erythroleukemia (MEL) cells as an expression system for nicotinic acetylcholine receptors." Thesis, University of Reading, 1997. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.389643.

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Passos, Débora Cristina Silva dos. "Ação biológica in vitro de tiossemicarbazonas derivadas de canfeno e limoneno em células de melanoma humano (SK-MEL-37)." Universidade Federal de Goiás, 2013. http://repositorio.bc.ufg.br/tede/handle/tede/3426.

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Melanoma is a type of cancer that arises from melanocytes and is notoriously resistant to radiation and chemotherapy. The thiosemicarbazones are synthetic compounds with marked biological properties such as antibacterial, antiviral, antiprotozoal and antitumor and previous studies have demonstrated cytotoxic activity against the human melanoma cells, so in this study, we evaluated the antiproliferative activity, the enzymatic activity of Caspases 2, 3, 6, 8, 9, the effect on the cell cycle gene expression levels of caspases 2, 3, 6, 8, 9, Apaf-1 and microscopic morphological changes in human melanoma cells (SK -MEL-37) twenty one monoterpene derived from natural thiosemicarbazone (-) - camphene: camphene, benzaldehyde, benzophenone, menthone, ethyl pyruvate, p-nitroacetophenone, pchloroacetophenone, p-methoxyacetophenone, p-methylacetophenone, p fluoracetofenona-phidroxiacetofena, furan, 3-methoxy-4-hydroxybenzaldehyde, p-fluorbenzaldehyde, 2- hydroxybenzaldehyde, cinnamic aldehyde, thiophene-2-carboxaldehyde, 1-H-imidazole-4- carboxaldehyde, tiossemicaroazida and six montoterpeno natural R-(+)-limonene: benzaldehyde, thiosemicarbazide, o-nitro, m-nitro, p-nitro, p-hydroxy and p-dimethylamino. The values found for the inhibitory concentration for 50% of cells (IC50) were between 12 μM and 55 μM. The percentage of cells in phase and in phase G0/G1 decreased SG2 / M increased after forty-eight hours of incubation with benzaldehyde thio-camphene, limonene thio-benzaldehyde, m-nitro, p-hydroxy and thiosemicarbazide increased indicating that the growth inhibitory effect might be also due to arrest of cells at S-G2/M phase. We observed increased activity of caspase 3 (m-nitro thio-limonene), 6 (camphene thio-benzaldehyde and p-hydroxy thio-limonene) and 8 (thio-benzaldehyde limonene). Late apoptotic features were detected in 62% of cells treated with benzaldehyde thio-camphene and morphological changes typical of apoptosis were visualized by fluorescence microscopy and scanning electron microscopy (SEM) after treatment with benzaldehyde thio-camphene chosen due to their low IC50 value (12 mM). It was observed gene expression of caspases 2, 3, 6, 8 and Apaf-1 in cells treated with benzaldehyde thio-camphene indicating the participation of these enzymes in the anti-proliferative effect observed. Our results indicate that the thiosemicarbazones derivatives can inhibit proliferation, regulate cell cycle, induce apoptosis of human melanoma cells (SK-MEL-37) and could be an candidate for future preclinical in vivo studies.
O melanoma é um tipo de câncer que surge nos melanócitos e é notoriamente resistente à radioterapia e quimioterapia. As tiossemicarbazonas são compostos sintéticos com marcantes propriedades biológicas tais como antibacteriana, antiviral, antiprotozoária e antitumoral e em estudos anteriores demonstraram ação citotóxica frente à celulas de melanoma humano, por isso, neste estudo, foi avaliada a atividade anti-proliferativa, a atividade enzimática das caspases 2, 3, 6, 8, 9, o efeito no ciclo celular, os níveis de expressão gênica das caspases 2, 3, 6, 8, 9, Apaf-1 e as alterações morfológicas por microscopia em células de melanoma humano (SK-MEL-37) de vinte e uma tiossemicarbazonas derivadas do monoterpeno natural (-)- canfeno: canfeno, benzaldeído, benzofenona, mentona, etil piruvato, acetofenona, pnitroacetofenona, p-cloroacetofenona, p-metoxiacetofenona, p-metilacetofenona, pfluoracetofenona, p-hidroxiacetofena, furano, 3-metóxi-4-hidroxibenzaldeído, pfluorbenzaldeído, 2-hidroxibenzaldeído, aldeído cinâmico, tiofeno-2-carboxialdeído, 1-Himidazol- 4-carboxialdeído, tiossemicaroazida, bem como seis do montoterpeno natural R-(+)- limoneno: benzaldeído, tiossemicarbazida, o-nitro, m-nitro, p-nitro, p-dimetilamino e phidróxi . Os valores encontrados para a concentração inibitória para 50% das células (IC50) situaram-se entre 12 μM e 55 μM. A porcentagem de células na fase G0/G1diminuiu e na fase SG2/M aumentou após quarenta e oito horas de incubação com o benzaldeído tio-canfeno, benzaldeído tio-limoneno, m-nitro, p-hidróxi e tiossemicarbazida, indicando que o efeito antiproliferativo observado pode ser devido a uma interrupção das células na fase SG2/M. Observou-se uma maior atividade de caspase 3 (m-nitro tio-limoneno), 6 (benzaldeído tiocanfeno e p-hidróxi tio-limoneno) e 8 (benzaldeído tio-limoneno). Características apoptóticas tardias foram detectados em 62% das células tratadas com benzaldeído tio-canfeno e as alterações morfológicas típicas de processo de apoptose foram visualizadas através da microscopia de fluorescência e de microscopia eletrônica de varredura (MEV) após tratamento com o benzaldeído tio-canfeno escolhido devido ao seu baixo valor de IC50 (12 μM). Observou-se a expressão gênica das caspases 2, 3, 6, 8 e o apaf-1 nas células tratadas com benzaldeído tio-canfeno indicando a participação dessas enzimas no efeito antiproliferativo observado. Os resultados indicam que as tiossemicarbazonas derivadas de canfeno e limoneno podem inibir a proliferação celular, regular o ciclo celular e induzir apoptose nas células de melanoma humano (SK-MEL-37), portanto, podem ser considerados candidatos para futuros ensaios pré-clínico in vivo.

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Hawke, David H. "Quantitative mass spectrometry: Proteomic analysis of differentiation of MEL cells treated with hexamethylene bisacetamide and 7-[N-(3-aminopropyl)amino] heptan-2-one." Scholarly Commons, 2004. https://scholarlycommons.pacific.edu/uop_etds/2691.

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Polyamines are small, polycationic molecules required for growth and development and found in all living cells. In this study, the effects of two polyamine analogues, hexamethylene bisacetamide (HMBA), a differentiation inducer, and 7-[N-(3-aminopropyl)amino] heptan-2-one (APAH), an inhibitor of N8-acetylspermidine deacetylase, were studied using quantitative proteomics and stable-isotopes. Two new technologies, isotope-coded affinity tags (ICAT) and quantification in fragment spectra using isobaric stable isotope reagents (iTRAQ) were employed and compared. Quantitative results of these experiments showed few changes in the type and level of proteins detected in whole-cell extracts. Proteins from three populations of cells were studied, control (untreated), HMBA-treated, and HMBA plus APAH treated cells. Some of the proteins that were differentially expressed in response to these agents include pyruvate kinase (PK), lactate dehydrogenase (LDH), mini-chromosome maintenance protein 3 (MCM3), and poly-rC binding protein. The proteins PK and LDH have been reported as possible cancer markers. Histone protein levels were significantly reduced on HMBA treatment, and substantially recovered with the addition of APAH. This finding was very convincing in the iTRAQ work, but invisible to the ICAT experiment, because of the lack of cysteine residues required for quantification in the ICAT methodology. Two proteins were elevated in the HMBA-APAH experiment compared to the other two, heterogeneous nuclear ribonuclear protein C1/C2 (HNRP C1/C2) and ubiquitin. Considering their unique functions, the up-regulation of these proteins suggests the involvement of internal ribosome entry and protein degradation in response to APAH. The results of the two technologies, ICAT and iTRAQ, were found to overlap, but were partly complementary.

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Morgado, Ana Sofia João. "Analysis of the intranuclear life of nonsense transcripts." Doctoral thesis, Faculdade de Ciências e Tecnologia, 2013. http://hdl.handle.net/10362/8798.

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Dissertação para obtenção do Grau de Doutor em Biologia, Especialidade de Biologia Molecular
Nonsense-mediated mRNA decay (NMD) is a quality control mechanism that detects and rapidly degrades mRNAs carrying premature translation-termination codons (PTCs). Mammalian NMD depends on both splicing and translation, and requires recognition of the premature stop codon by the cytoplasmic ribosomes. Surprisingly, some published data have suggested that nonsense codons may also affect the nuclear metabolism of the nonsense-mutated transcripts. Therefore, we hypothesized that human β-globin transcripts sensitive to NMD could have a singular subcellular localization and processing state in mammalian cells nuclei. To determine if PTCs could influence nuclear events, we have established mouse erythroleukemia (MEL) cell lines stably transfected with wild-type or PTC-containing human β-globin genes. Subsequently, we analyzed the accumulation of NMD-competent β-globin transcripts versus wild-type counterparts using two different approaches: visualization of transcripts localization by fluorescence in situ hybridization (FISH); and quantification of pre-mRNA steady-state levels by ribonuclease protection assays (RPA) and reverse transcription-coupled quantitative polymerase chain reaction (RT-qPCR). FISH analysis shows that MEL cells stably expressing PTC-containing β-globin transcripts present a marked tendency to display an abnormal speckled-like pattern of localization in the nucleus. However, in addition to the presence of the PTC, other effectors may act on the β-globin transcripts localization, as some wild-type β-globin MEL cells presented this abnormal FISH phenotype as well. On the other hand, our analyses by RPA and RT-qPCR clearly show that β- -globin pre-mRNAs carrying NMD-competent PTCs, but not those containing a NMD-resistant PTC, exhibit a significant decrease in their steady-state levels relatively to the wild-type or to a missense-mutated β-globin pre-mRNA. Conversely, in non-erythroid HeLa cells, human β-globin pre-mRNAs carrying NMD-competent PTCs accumulate at normal levels. Half-life analysis of these pre-mRNAs in MEL cells demonstrate that their low steady-state levels do not reflect significantly lower pre-mRNA stabilities when compared to the normal control. Furthermore, our results also provide evidence that the relative splicing efficiencies of intron 1 and 2 are unaffected. In conclusion, our set of data highlights potential nuclear pathways that induce a selective downregulation of PTC-containing β-globin pre-mRNA in MEL cells, albeit not affecting their stability or splicing effectiveness. These specialized nuclear pathways, which may act in concert with the general NMD mechanism, might discriminate the NMD-sensitive transcripts as abnormal in a promoter- and/or cell line-specific manner, probably to obtain optimal NMD activity.
Fundação para a Ciência e Tecnologia - (SFRH/BD/31920/2006); financial support [Centro de Investigação em Genética Molecular Humana (CIGMH) and Center for Biodiversity, Functional and Integrative Genomics (BioFIG)]

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Valente, Sabrina <1980&gt. "Vascular wall stem cells. Selection and conditioning of progenitors useful for cell therapy. A pathological case study." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2010. http://amsdottorato.unibo.it/2857/.

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The arterial wall contains MSCs with mesengenic and angiogenic abilities. These multipotent precursors have been isolated from variously-sized human adult segments, belying the notion that vessel wall is a relatively quiescent tissue. Recently, our group identified in normal human arteries a vasculogenic niche and subsequently isolated and characterized resident MSCs (VW-MSCs) with angiogenic ability and multilineage potential. To prove that VW-MSCs are involved in normal and pathological vascular remodeling, we used a long-term organ culture system; this method was of critical importance to follow spontaneous 3-D vascular remodeling without any influence of blood cells. Next we tried to identify and localize in situ the VW-MSCs and to understand their role in the vascular remodeling in failed arterial homografts. Subsequently, we isolated this cell population and tested in vitro their multilineage differentiation potential through immunohistochemical, immunofluorescence, RT-PCR and ultrastructural analysis. From 25-30cm2 of each vascular wall homograft sample, we isolated a cell population with MSCs properties; these cells expressed MSC lineage molecules (CD90, CD44, CD105, CD29, CD73), stemness (Notch-1, Oct-4, Sca-1, Stro-1) and pericyte markers (NG2) whilst were negative for hematopoietic and endothelial markers (CD34, CD133, CD45, KDR, CD146, CD31 and vWF). MSCs derived from failed homografts (H-MSCs) exhibited adipogenic, osteogenic and chondrogenic potential but scarce propensity to angiogenic and leiomyogenic differentiation. The present study demonstrates that failed homografts contain MSCs with morphological, phenotypic and functional MSCs properties; H-MSCs are long-lived in culture, highly proliferating and endowed with prompt ability to differentiate into adipocytes, osteocytes and chondrocytes; compared with VW-MSCs from normal arteries, H-MSCs show a failure in angiogenic and leiomyogenic differentiation. A switch in MSCs plasticity could be the basis of pathological remodeling and contribute to aneurysmal failure of arterial homografts. The study of VW-MSCs in a pathological setting indicate that additional mechanisms are involved in vascular diseases; their knowledge will be useful for opening new therapeutic options in cardiovascular diseases.

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Curradi, Giacomo <1977&gt. "Airway Basal Cell Vascular Endothelial Growth Factor-mediated Cross-Talk Regulates Endothelial Cell Dependent Growth Support of Human Airway Basal Cells." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2013. http://amsdottorato.unibo.it/5925/.

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The human airway epithelium is a pseudostratified heterogenous layer comprised of cili-ated, secretory, intermediate and basal cells. As the stem/progenitor population of the airway epi-thelium, airway basal cells differentiate into ciliated and secretory cells to replenish the airway epithelium during physiological turnover and repair. Transcriptome analysis of airway basal cells revealed high expression of vascular endothelial growth factor A (VEGFA), a gene not typically associated with the function of this cell type. Using cultures of primary human airway basal cells, we demonstrate that basal cells express all of the 3 major isoforms of VEGFA (121, 165 and 189) but lack functional expression of the classical VEGFA receptors VEGFR1 and VEGFR2. The VEGFA is actively secreted by basal cells and while it appears to have no direct autocrine function on basal cell growth and proliferation, it functions in a paracrine manner to activate MAPK signaling cascades in endothelium via VEGFR2 dependent signaling pathways. Using a cytokine- and serum-free co-culture system of primary human airway basal cells and human endothelial cells revealed that basal cell secreted VEGFA activated endothelium to ex-press mediators that, in turn, stimulate and support basal cell proliferation and growth. These data demonstrate novel VEGFA mediated cross-talk between airway basal cells and endothe-lium, the purpose of which is to modulate endothelial activation and in turn stimulate and sustain basal cell growth.
I risultati preliminari dello studio suggeriscono che il vascular endothelial growth factor A(VEGFA) e’ attivamente secreto dalle cellule basali dell’epitelio bronchiale e svolge una funzione paracrina nell’attivazione della cascata delle mitogen-activated protein kinases (MAPKs) nelle cellule endoteliali mediata dal VEGF receptor type 2. Utilizzando un sistema di co-coltura di cellule basali primarie delle vie aeree umane con cellule endoteliali umane, abbiamo mostrato come il VEGFA secreto dalle cellule basali sia in grado di attivare le cellule endoteliale che a loro volta, esprimono mediatori capaci di stimolare e sostenere la proliferazione delle cellule basali stesse. Questi dati dimostrano un cross-talk mediato dal rilascio di VEGFA tra le cellule basali dell’epitelio bronchiale e l’endotelio, il cui scopo è di modulare l'attivazione endoteliale e, a sua volta stimolare e sostenere la crescita delle cellule basali.

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Avanzi, Simone <1982&gt. "Cell Host-Microbe Interactions: Turning Pathogen Mechanisms Into Cell's Advantages." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2013. http://amsdottorato.unibo.it/5759/.

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Host-Pathogen Interaction is a very vast field of biological sciences, indeed every year many un- known pathogens are uncovered leading to an exponential growth of this field. The present work lyes between its boundaries, touching different aspects of host-pathogen interaction: We have evaluate the permissiveness of Mesenchimal Stem cell (FM-MSC from now on) to all known human affecting herpesvirus. Our study demonstrate that FM-MSC are full permissive to HSV1, HSV2, HCMV and VZV. On the other hand HHV6, HHV7, EBV and HHV8 are susceptible, but failed to activate a lytic infection program. FM-MSC are pluripotent stem cell and have been studied intensely in last decade. FM-MSC are employed in some clinical applications. For this reason it is important to known the degree of susceptibility to transmittable pathogens. Our atten- tion has then moved to bacterial pathogens: we have performed a proteome-wide in silico analy- sis of Chlamydiaceae family, searching for putative Nuclear localization Signal (NLS). Chlamy- diaceae are a family of obligate intracellular parasites. It’s reasonably to think that its members could delivered to nucleus effector proteins via NLS sequences: if that were the case the identifi- cation of NLS carrying proteins could open the way to therapeutic approaches. Our results strengthen this hypothesis: we have identified 72 protein bearing NLS, and verified their func- tionality with in vivo assays. Finally we have conceived a molecular scissor, creating a fusion protein between HIV-1 IN protein and FokI catalytic domain (a deoxyexonuclease domain). Our aim is to obtain chimeric enzyme (trojIN) which selectively identify IN naturally occurring target (HIV LTR sites) and cleaves subsequently LTR carrying DNA (for example integrated HIV1 DNA). Our preliminary results are promising since we have identified trojIN mutated version capable to selectively recognize LTR carrying DNA in an in vitro experiments.

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Wang, Aibo. "Phosphorylation of Nur77 by MEK-ERK-RSK cascade induces mitochondrial translocation and apoptosis in T cells." Amherst, Mass. : University of Massachusetts Amherst, 2009. http://scholarworks.umass.edu/dissertations/AAI3372283/.

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Gallerani, Giulia <1986&gt. "Circulating Tumor Cells Investigation in Esophageal Cancer." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2016. http://amsdottorato.unibo.it/7343/.

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Background: An increasing number of studies have established that circulating tumor cells (CTCs) are a heterogeneous population in which cells have different degrees of metastatic potential mainly due to epithelial-mesenchymal transition (EMT). This study aimed to assess the feasibility of circulating esophageal cancer cells identification, characterization and to evaluate their prognostic value in esophageal cancer Methods: In order to closely mimic CTC heterogeneity, MC10A and HMEL cell lines differently forced in EMT are used. Single cell analysis was conducted by DEPArray. We assigned a specific phenotypic tag to each potential CTC population: Epithelial-tag, Mesenchymal/stem-tag. We evaluated the basal cell phenotype and after EMT induction. Subsequently, the Grab all assay was performed on peripheral blood samples from patients with esophageal cancer. This feasibility study enrolled 11 patients (4 M1, 7 M0). Analyses were conducted on 3 peripheral blood samples (15/20 ml) per patients. Blood samples from non-metastatic patients were taken before and after primary neo-adjuvant therapy and after secondary treatment (surgery). Samples from metastatic patients were taken before and after first line therapy and after second line therapy. CTCs were identified and sorted singly by DEPArray system. Results: The assay was able to detect the phenotypic changes in cell lines mimicking CTC heterogeneity. Before therapy, CTCs were found in 3/4 metastatic patients. Of the 7 non-metastatic patients, 3 were positive for CTCs before therapy. Examining CTC status at different clinical time points, it was possible to suggest a correlation between the presence of CTCs and disease progression. Conclusions: Data showed that the assay is feasible, capable to analyze the phenotypic tags by DEPArray using a multiple staining without aspecific signals. Experiments carried out from both metastatic/non metastatic cancer patients showed the ability of the Grab-allassay to identify subpopulations of CTCs with different epithelial/stem/mesenchymal or hybrid phenotypes potentially related to disease progression.

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Foroni, Laura <1978&gt. "Resident angiogenic mesenchymal stem cells from multiorgan donor thoracic aortas." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2008. http://amsdottorato.unibo.it/976/.

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Stem cells are one of the most fascinating areas of biology today, and since the discover of an adult population, i.e., adult Stem Cells (aSCs), they have generated much interest especially for their application potential as a source for cell based regenerative medicine and tissue engineering. aSCs have been found in different tissues including bone marrow, skin, intestine, central nervous system, where they reside in a special microenviroment termed “niche” which regulate the homeostasis and repair of adult tissues. The arterial wall of the blood vessels is much more plastic than ever before believed. Several animal studies have demonstrated the presence of cells with stem cell characteristics within the adult vessels. Recently, it has been also hypothesized the presence of a “vasculogenic zone” in human adult arteries in which a complete hierarchy of resident stem cells and progenitors could be niched during lifetime. Accordingly, it can be speculated that in that location resident mesenchymal stem cells (MSCs) with the ability to differentiate in smooth muscle cells, surrounding pericytes and fibroblasts are present. The present research was aimed at identifying in situ and isolating MSCs from thoracic aortas of young and healthy heart-beating multiorgan donors. Immunohistochemistry performed on fresh and frozen human thoracic aortas demonstrated the presence of the vasculogenic zone between the media and the adventitial layers in which a well preserved plexus of CD34 positive cells was found. These cells expressed intensely HLA-I antigens both before and after cryopreservation and after 4 days of organ cultures remained viable. Following these preliminary results, we succeeded to isolate mesenchymal cells from multi-organ thoracic aortas using a mechanical and enzymatic combined procedure. Cells had phenotypic characteristics of MSC i.e., CD44+, CD90+, CD105+, CD166+, CD34low, CD45- and revealed a transcript expression of stem cell markers, e.g., OCT4, c-kit, BCRP-1, IL6 and BMI-1. As previously documented using bone marrow derived MSCs, resident vascular wall MSCs were able to differentiate in vitro into endothelial cells in the presence of low-serum supplemented with VEGF-A (50 ng/ml) for 7 days. Under the condition described above, cultured cells showed an increased expression of KDR and eNOS, down-regulation of the CD133 transcript, vWF expression as documented by flow cytometry, immunofluorescence, qPCR and TEM. Moreover, matrigel assay revealed that VEGF induced cells were able to form capillary-like structures within 6 hours of seeding. In summary, these findings indicate that thoracic aortas from heart-beating, multi-organ donors are highly suitable for obtaining MSCs with the ability to differentiate in vitro into endothelial cells. Even though their differentiating potential remains to be fully established, it is believed that their angiogenic ability could be a useful property for allogenic use. These cells can be expanded rapidly, providing numbers which are adequate for therapeutic neovascularization; furthermore they can be cryostored in appropriate cell banking facilities for later use.

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Romano, Marco <1985&gt. "In vitro characterisation and expansion of human regulatory T cells for their in vivo application in the induction of tolerance in haematopoietic stem cell and solid organ transplantation." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2015. http://amsdottorato.unibo.it/6784/.

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Solid organ transplantation (SOT) is considered the treatment of choice for many end-stage organ diseases. Thus far, short term results are excellent, with patient survival rates greater than 90% one year post-surgery, but there are several problems with the long term acceptance and use of immunosuppressive drugs. Hematopoietic Stem Cells Transplantation (HSCT) concerns the infusion of haematopoietic stem cells to re-establish acquired and congenital disorders of the hematopoietic system. The main side effect is the Graft versus Host Disease (GvHD) where donor T cells can cause pathology involving the damage of host tissues. Patients undergoing acute or chronic GvHD receive immunosuppressive regimen that is responsible for several side effects. The use of immunosuppressive drugs in the setting of SOT and GvHD has markedly reduced the incidence of acute rejection and the tissue damage in GvHD however, the numerous adverse side effects observed boost the development of alternative strategies to improve the long-term outcome. To this effect, the use of CD4+CD25+FOXP3+ regulatory T cells (Treg) as a cellular therapy is an attractive approach for autoimmunity disease, GvHD and limiting immune responses to allograft after transplantation. Treg have a pivotal role in maintaining peripheral immunological tolerance, by preventing autoimmunity and chronic inflammation. Results of my thesis provide the characterization and cell processing of Tregs from healthy controls and patients in waiting list for liver transplantation, followed by the development of an efficient expansion-protocol and the investigation of the impact of the main immunosuppressive drugs on viability, proliferative capacity and function of expanded cells after expansion. The conclusion is that ex vivo expansion is necessary to infuse a high Treg dose and although many other factors in vivo can contribute to the success of Treg therapy, the infusion of Tregs during the administration of the highest dose of immunosuppressants should be carefully considered.

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Hodge, Jacob G. "Regulation of the MEK/ERK signaling cascade by ADAM12 in triple-negative breast cancer cells." Thesis, Kansas State University, 2016. http://hdl.handle.net/2097/35228.

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Master of Science
Biochemistry and Molecular Biophysics Interdepartmental Program
Anna Zolkiewska
Mitogen-activated protein kinase (MAPK) signaling plays an important role in the proliferation, survival, and therapy resistance of breast cancer cells. Two important protein kinases involved in the MAPK pathway are MEK and ERK. The MEK/ERK signaling cascade can be stimulated by activation of the epidermal growth factor receptor (EGFR) upon binding of EGF-like ligands, which are released from cells by ADAM proteases. EGFR is frequently overexpressed in triple-negative breast cancer (TNBC), a particularly aggressive form of breast cancer. Our analysis of clinical data revealed that high expression of ADAM12, but not other ADAMs, in TNBC is associated with poor patient survival. Thus, we hypothesized that ADAM12 plays a critical role in the progression of TNBC, possibly by stimulating MEK/ERK activity in an EGFR-dependent manner. To test this hypothesis, ADAM12 was knocked-down (KD) in SUM159PT TNBC cells, which express high levels of the endogenous ADAM12 protein. An antibody array assay indicated a significant decrease in the activation of the MAPK pathway in SUM159PT cells after ADAM12 KD. The decrease in MAPK activity was further confirmed by Western blotting using phospho-MEK and phospho-ERK specific antibodies. Additionally, conditioned media from ADAM12-deficient SUM159PT cells failed to support the survival of MCF10A cells, suggesting that ADAM12 KD reduced the release of pro-survival growth factors from SUM159PT cells. Based upon this data, we propose that ADAM12 is a novel regulator of the MAPK pathway and a potential therapeutic target in breast cancer.

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15

Capelli, Irene <1979&gt. "A step forwards in immunomodulation and immunetolerance knowledge. HLA-G expression in co-cultures of peripheral blood mononuclear cells and stem cells after in vitro NGAL stimulation." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2013. http://amsdottorato.unibo.it/5795/.

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NGAL (Neutrophil Gelatinase-associated Lipocalin ) is a protein of lipocalin superfamily. Recent literature focused on its biomarkers function in several pathological condition (acute and chronic kidney damage, autoimmune disease, malignancy). NGAL biological role is not well elucidated. Several are the demonstration of its bacteriostatic role. Recent papers have indeed highlight NGAL role in NFkB modulation. The aim of this study is to understand whether NGAL may exert a role in the activation (modulation) of T cell response through the regulation of HLA-G complex, a mediator of tolerance. From 8 healthy donors we obtained peripheral blood mononuclear cells (PBMCs) and we isolated by centrifugation on a Ficoll gradient. Cells were then treated with four concentrations of NGAL (40-320 ng/ml) with or without iron. We performed flow cytometry analysis and ELISA test. NGAL increased the HLA-G expression on CD4+ T cells, with an increasing corresponding to the dose. Iron effect is not of unique interpretation. NGAL adiction affects regulatory T cells increasing in vitro expansion of CD4+ CD25+ FoxP3+ cells. Neutralizing antibody against NGAL decreased HLA-G expression and reduced significantly CD4+ CD25+ FoxP3+ cells percentage. In conclusion, we provided in vitro evidence of NGAL involvement in cellular immunity. The potential role of NGAL as an immunomodulatory molecule has been evaluated: it has been shown that NGAL plays a pivotal role in the induction of immune tolerance up regulating HLA-G and T regulatory cells expression in healthy donors. As potential future scenario we highlight the in vivo role of NGAL in immunology and immunomodulation, and its possible relationship with immunosuppressive therapy efficacy, tolerance induction in transplant patients, and/or in other immunological disorders.
Ngal (Neutrophil Gelatinase-associated Lipocalin ) è una proteina appartenente alla famiglia delle lipocaline verso cui la recente letteratura ha mostrato una notevole attenzione, soprattutto in quanto biomarcatore in alcune condizioni patologiche (danno renale acuto e cronico, patologie autoimmuni, neoplasie). Il ruolo biologico di NGAL non è però ancora del tutto compreso. Numerose sono le dimostrazioni della sua azione batteriostatica. Recenti lavori hanno inoltre evidenziato un ruolo di NGAL nella modulazione di NFkB. Nessun lavoro ha valutato il ruolo di NGAL nell’immunità umorale. Lo scopo dello studio è quello di capire se NGAL possa esercitare un ruolo di attivazione (modulazione) della risposta T cellulare attraverso la regolazione del complesso HLA-G, un mediatore di tolleranza. Cellule mononucleate da sangue periferico (PBMCs) sono state ottenute da 8 donatori sani dopo consenso informato e isolate tramite centrifugazione (Ficoll). PBMC sono poi state trattate con 4 concentrazioni crescenti di NGAL (da 40 a 320 ng/mL), associate o meno a ferro e analizzate con tecnica fluorimetrica ed elisa.Alle analisi eseguite NGAL stimola l’espressione di HLA-G sulle cellule T CD4+ con un andamento dose dipendente. L’effetto del ferro sull’espressione di HLA-G non è di univoca interpretazione.Inoltre L’aggiunta di NGAL in vitro modifica il pattern di espressione delle cellule T, aumentando la popolazione delle cellule CD4+ CD25+ FoxP3. L’utilizzo di anticorpi anti NGAL limita l’espressione di HLA-G e diminuisce significativamente la percentuale di CD4+ CD25+ FoxP3+ . In conclusione abbiamo mostrato un coinvolgimento di NGAL nell’immunità cellulare. Valutando il ruolo di NGAL come molecola immunomodulatoria, abbiamo mostrato che NGAL gioca un ruolo chiave neell’immunotolleranza aumentando l’espressione di HLA-G e cellule T regolatorie nei donatori sani. Un possibilAs potential future scenario applicativo di tale studio riguarda l’utilizzo in vivo di NGAL nell’immunomodulazione dei pazienti sottoposti a trapianto o affetti da patologie autoimmuni.

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La, Morgia Chiara <1977&gt. "Melanopsin Retinal Ganglion Cells: relevance to circadian rhythms and sleep in neurodegeneration." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2012. http://amsdottorato.unibo.it/4796/.

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In this PhD thesis 3 projects were addressed focusing on the melanopsin retinal ganglion cells (mRGCs) system and its relevance for circadian rhythms and sleep in neurodegeneration. The first project was aimed at completing the characterization of mRGCs system in hereditary optic neuropathies (LHON and DOA). We confirmed that mRGCs are relatively spared also in post-mortem retinal specimens of a DOA case and pupillometric evaluation of LHON patients showed preservation of the pupillary light reflex, with attenuated responses compared to controls. Cell studies failed to indicate a protective role exerted by melanopsin itself. The second project was aimed at characterizing the possible occurrence of optic neuropathy and rest-activity circadian rhythm dysfunction in Alzheimer (AD) and Parkinson disease (PD), as well as, at histological level, the possible involvement of mRGCs in AD. OCT studies demonstrated a subclinical optic neuropathy in both AD and PD patients, with a different pattern involving the superior and nasal quadrants in AD and the temporal quadrant in PD. Actigraphic studies demonstrated a tendency towards an increased intradaily variability (IV) and reduced relative amplitude (RA) of rest-activity circadian rhythm in AD and a significant increased IV a reduced RA in PD. Immunohistochemical analysis of post-mortem retinal specimens and optic nerve cross-sections of neuropathologically confirmed AD cases demonstrated a significant loss of mRGCs and a nearly significant loss of axons in AD compared to controls. The mRGCs were affected in AD independently from age and magnitude of axonal loss. Overall these results suggest a role of the mRGCs system in the pathogenesis of circadian dysfunction in AD. The third project was aimed at evaluating the possible association between a single nucleotide polymorphism of the OPN4 gene and chronotype or SAD, failing to find any significant association with chronotype, but showing a non-significant increment of TT genotype in SAD.

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Yagi, Rieko. "Bcl-2 Regulates Chondrocyte Phenotype Through MEK-ERK1/2 Pathway; Relevance to Osteoarthritis and Cartilage Biology." [Kent, Ohio] : Kent State University, 2005. http://rave.ohiolink.edu/etdc/view?acc%5Fnum=kent1118329494.

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Thesis (Ph.D.)--Kent State University, 2005.
Title from PDF t.p. (viewed Sept. 5, 2006). Advisor: Walter E. Horton. Keywords: chondrocytes; osteoarthritis; Sox9; Bcl-2; MEK-ERK 1/2. Includes bibliographical references (p. 91-106).

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18

Frigault, Melanie M. (Melanie Mae) 1979. "The role of the Gab family of docking proteins in Met mediated membrane ruffle formation /." Thesis, McGill University, 2008. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=115673.

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In response to extra-cellular cues, cells activate signal transduction pathways to elicit a biological response. Cell surface growth factor receptors such as the Met receptor tyrosine kinase (RTK) activate signals that result in cellular proliferation, survival, migration, as well as epithelial morphogenesis. In order for signal transduction to occur, docking proteins are recruited to the activated RTK, become phosphorylated on tyrosine residues, which then serve as docking sites for the recruitment of other signaling proteins. Docking proteins function to diversify the signal by assembling multi-protein complexes. The Gab1 docking protein is the most tyrosine phosphorylated protein upon Met receptor activation and is required for Met mediated signaling and biology.
Gab1 belongs to a family of docking proteins including the highly related Gab2 protein. Gab1 promotes signals for epithelial morphogenesis downstream of the Met receptor, however Gab2 is unable to do so. Insertion of the Gab1 Met binding Motif (MBM) which confers direct binding to the Met receptor, as well as membrane targeting of Gab2 is sufficient to switch the capacity of Gab2 to activate the morphogenic program, cell scatter and lamellipodia formation. This is achieved via activation of sustained signaling pathways, and redistribution of the Gab protein, and associated molecules to sites of lamellipodia formation at the peripheral edge of the cell.
Activation of the Met RTK, promotes the formation of dorsal ruffles on the apical surface of epithelial cells. The Met receptor, Gab1 and Gab1 associated molecules Shp2, Crk, and p8S subunit of PI3K, are localized to these structures, however only the Gab1erk complex is required to drive dorsal ruffle formation. Gab1 is required for Met induced dorsal ruffles as well as downstream the PDGF and EGF RTKs. These are a signaling micro-environment which results in enhanced receptor degradation. Inhibition or enhancement of Met mediated dorsal ruffle formation correlates with receptor stability.
Dorsal ruffle formation downstream of Met requires the enzymatic activity of PI3K and PLCgamma, both enzymes that metabolize PIP2, and form complexes with Gab1 downstream of Met. PLCgamma and the PIP3 lipid product of PI3K are co-localized with Gab1 in dorsal ruffles. Gab1 engages with elements of the cytoskeleton, actin and cortactin, providing a link between growth factor signaling and remodeling of the actin cytoskeleton. Gab1 is localized to membrane protrusions of the basal surface in organoid cultures and is required for actin protrusions of the basal surface of breast cancer cells.

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19

Tucker, Catherine Amanda. "Targeted therapies in mantle cell lymphoma." Thesis, University of British Columbia, 2008. http://hdl.handle.net/2429/922.

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Mantle cell lymphoma (MCL) is characterized by the presence of the t(11 ;14)(g13 ;g32) translocation which results in cyclin Dl over-expression. MCL is one of the most difficult lymphoproliferative disorders to manage with a median survival rate of 43 months from diagnosis. The poor prognosis associated with MCL is due in large part to its late classification as a separate clinical entity leading to a dearth in available pre-clinical models. The specific objectives of the research described in this thesis were (1) to establish MCL preclinical models of disease and (2) to evaluate deregulated cell signaling pathways in MCL that can impact treatment response. Pre-clinical models of MCL were established from pre-existing cell lines containing the t(11 ;14)(g13 ;g32). These cell lines were previously misclassified because they were developed prior to the classification of MCL as a distinct lymphoma subtype. With the establishment of MCL models, deregulated cell signaling pathways in MCL and response to different treatment strategies were investigated. These included an investigation of the cell signaling pathways activated in bcl-2 over-expressing MCL cells that were treated with oblimersen; a molecular gene silencing strategy that effectively suppresses bcl-2 in vitro and in vivo. Silencing bcl-2 provided insight into which pathways were influenced by bcl-2 over-expression in MCL. More specifically loss of cyclin D1, NF-KB, p53, bax and p27 were observed following bcl-2 silencing. Additional studies investigated how abnormal expression of CD40/CD40L and Fas/FasL along with bcl-2 family members contributes to B cell clonal expansion and influences Rituximab-mediated cell death in MCL models. Rituximab is a chimeric monoclonal antibody targeted against B cells and both Rituximab-sensitive and insensitive MCL models were defined. An abnormally high expression of bcl-2, bcl-x L, mcl-1, CD40/CD40L and Fas were observed in all MCL cells, as well as high levels of soluble FasL, capable of blocking Fas-mediated apoptosis. These deregulated pathways were associated with response to Rituximab treatment in a sensitive MCL model. These studies demonstrated some of the key pathways associated with treatment response in MCL, and the establishment of well characterized MCL models enables us to continue to explore new treatment strategies currently being studied in other lymphomas.

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20

Hamilton, William. "Functional and biochemical analysis of ERK2 in mouse embryonic stem cells." Thesis, University of Edinburgh, 2011. http://hdl.handle.net/1842/5774.

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The ERK-MAPK pathway is a dynamic signaling module, conserved across Eukarya, and capable of processing a myriad of environmental and cellular signals. It has been implicated in controlling important cell fate decisions in many cell types and species. In mES cells, growth factor activation of the ERK-MAPK pathway is involved in the earliest stages of lineage segregation, however very little is currently known about the mechanism by which this is accomplished. Taking a loss-of-function gene targeting approach I have reexamined the relative contribution of ERK2 activity to FGF-ERK signaling. Although ERK2 depletion results in an attenuation of the combined ERK1/2 activity, this is compensated for by the hyperactivation of the remaining ERK1 isozyme. Normal ERK1/2 function can be restored to ERK2 deficient cells by transgenic expression of either ERK1 or ERK2, indicating a degree of functional redundancy between both isoforms. When subjected to the appropriate cues, lineage commitment proceeded normally in ERK2 deficient cells, however increased self-renewal was observed under standard culture conditions. Several attempts were made to further probe ERK1/2 function by siRNA depletion, and dominant negative inhibition of ERK1 in Erk2 knockout cells, however both approaches failed to provide further insight. Furthermore, taking a candidate approach, the role of Srf, a canonical target of ERK1/2 signaling, was examined. Initial experiments indicated a role for SRK in neural differentiation, however due to issues of culture adaptation and instability in several cell lines it was not possible to conclude this line of research within the time frame of this thesis. IP-MS/MS analysis identified several proteins known to interact with ERK2 and indicated an involvement in nuclear pore function through TPR as well as transcriptional and translational regulation through RSK proteins. Moreover, this study identified DUSP6 and DUSP9 as the primary induced dual specificity phosphatases that regulate ERK2 activity in mES cells. To further probe the functional significance of the ERK:p90RSK interaction I examined a mES cell line genetically depleted for PDK1, a crucial regulator of p90RSK function. This cell line exhibits no detectable p90RSK activity, however in contrast to studies in other cell lines, p90RSK activity is dispensable for mitogen-induced cFos expression in mES cells. Subsequent experiments demonstrated a requirement for PDK1 activity in either the specification or maintenance of mES cell derived neurons. Further analysis indicated that p90RSK may be involved in a negative feedback loop regulating ERK1/2 activity, and if so may represent a point whereby ERK1/2 activity can be manipulated. To examine this I determined the effect pharmacological inhibition of p90RSK has on ERK1/2 activity and self-renewal using a novel p90Rsk inhibitor, BI-D1870. Although treatment with BI-D1870 correlated with enhanced ERK1/2 phosphorylation, the offtarget effects this molecule exhibits made it impossible to draw any firm conclusions from these experiments. Overall this study has demonstrated a degree of redundancy between ERK1/2 isozymes in mES cells. It has highlighted the complex nature of ERK1/2 regulation as well as the robustness of this pathway to perturbations in ERK dose. Furthermore, it has underscored some of the common pitfalls encountered when studying differentiation phenotypes in mES cells. Although this study failed to highlight anything more than a coincidental relationship between ERK1/2 activity and self-renewal capacity of mES cells, it has helped to highlight some important behavioral characteristics of the FGF-MAPK pathway in mES cells and provide a platform for further study.

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Chiereghin, Angela <1982&gt. "Epstein-Barr Virus-Related B Cell Lymphoproliferative Disorder After Hematopoietic Stem Cell Transplantation." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2016. http://amsdottorato.unibo.it/7611/.

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In 51 (33 adult-18 paediatric) allogeneic hematopoietic stem cell transplant recipients we aimed to evaluate: i) the incidence of EBV infection and potential risk factors; ii) the suitability of whole blood (WB) as clinical specimen to monitor the risk of patients to develop EBV-related post-transplant lymphoproliferative disorder (EBV-PTLD); iii) the clinical utility of combined virological-immunological monitoring; iv) the management of infection and the incidence of EBV-PTLD. Quantitative real-time PCR assay was performed on WB samples for all patients. EBV-DNA quantification also in peripheral blood mononuclear cells (PBMCs) samples was adopted for actively EBV infected patients. Immunological monitoring of infection was performed by Enzyme-linked ImmunoSPOT assay evaluating the EBV-specific cell-mediated immunity (CMI). The incidence of EBV infection was 51% and the frequency of EBV-PTLD was 3.9%. Reduced-intensity conditioning (RIC) in combination with in vivo T-cell depletion was associated with higher frequencies of infection (P=0.036). A significant correlation (P<0.001) between EBV-DNA levels in WB and PBMC samples was obtained in adult (r=0.787) and paediatric (r=0.976) patients. A similar kinetics of EBV-DNA in blood compartments was observed. Clinically, both specimen types appeared to be equally informative. The lack of EBV-specific CMI during/after active infection was associated with a higher median peak level of EBV-DNA in WB (P=0.013) and a greater severity of infection. The 54.5% of the patients without EBV-specific CMI needed anti-CD20 therapy and the 27.2% developed EBV-related complications, including a lethal PTLD. All patients with EBV-specific CMI controlled EBV replication and were asymptomatic. WB proved to be a suitable clinical specimen to monitor the risk of patients to develop EBV-related complications. RIC combined with in vivo T-cell depletion is a risk factor for the development of infection. EBV-specific CMI is a critical determinant in controlling the infection and consequently the EBV-related complications. Combined virological-immunological monitoring could improve the management of infection.

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Pacilli, Annalisa <1983&gt. "Characterization of vascular wall progenitor cells and their role in therapeutic angiogenesis and Monckeberg's sclerosis." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2011. http://amsdottorato.unibo.it/3511/.

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Recently, the existence of a capillary-rich vasculogenic zone has been identified in adult human arteries between the tunica media and adventitia; in this area it has been postulated that Mesenchymal Stem Cells (MSCs) may be present amidst the endothelial progenitors and hematopoietic stem cells. This hypothesis is supported by several studies claiming to have found the in vivo reservoir of MSCs in post-natal vessels and by the presence of ectopic tissues in the pathological artery wall. We demonstrated that the existence of multipotent progenitors is not restricted to microvasculature; vascular wall resident MSCs (VW-MSCs) have been isolated from multidistrict human large and middle size vessels (aortic arch, thoracic aorta and femoral artery) harvested from healthy multiorgan donors. Each VW-MSC population shows characteristics of embryonic-like stem cells and exhibits angiogenic, adipogenic, chondrogenic and leiomyogenic potential but less propensity to osteogenic ifferentiation. Human vascular progenitor cells are also able to engraft, differentiate into mature endothelial cells and support muscle function when injected in a murine model of hind limb ischemia. Conversely, VW-MSCs isolated from calcified femoral arteries display a good response to osteogenic commitment letting us to suppose that VW-MSCs could have an important role in the onset of vascular pathologies such as Mönckeberg sclerosis. Taken together these results show two opposite roles of vascular progenitor cells and underline the importance of establishing their in vivo pathological and regenerative potential to better understand pathological events and promote different therapeutic strategies in cardiovascular research and clinical applications.

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Olivi, Elena <1982&gt. "Adipose-derived stem cells and tissue revascularization: enhancing islet survival and performance for diabetes care." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2013. http://amsdottorato.unibo.it/5603/.

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Pancreatic islet transplantation represents a fascinating procedure that, at the moment, can be considered as alternative to standard insulin treatment or pancreas transplantation only for selected categories of patients with type 1 diabetes mellitus. Among the factors responsible for leading to poor islet engraftment, hypoxia plays an important role. Mesenchymal stem cells (MSCs) were recently used in animal models of islet transplantation not only to reduce allograft rejection, but also to promote revascularization. Currently adipose tissue represents a novel and good source of MSCs. Moreover, the capability of adipose-derived stem cells (ASCs) to improve islet graft revascularization was recently reported after hybrid transplantation in mice. Within this context, we have previously shown that hyaluronan esters of butyric and retinoic acids can significantly enhance the rescuing potential of human MSCs. Here we evaluated whether ex vivo preconditioning of human ASCs (hASCs) with a mixture of hyaluronic (HA), butyric (BU), and retinoic (RA) acids may result in optimization of graft revascularization after islet/stem cell intrahepatic cotransplantation in syngeneic diabetic rats. We demonstrated that hASCs exposed to the mixture of molecules are able to increase the secretion of vascular endothelial growth factor (VEGF), as well as the transcription of angiogenic genes, including VEGF, KDR (kinase insert domain receptor), and hepatocyte growth factor (HGF). Rats transplanted with islets cocultured with preconditioned hASCs exhibited a better glycemic control than rats transplanted with an equal volume of islets and control hASCs. Cotransplantation with preconditioned hASCs was also associated with enhanced islet revascularization in vivo, as highlighted by graft morphological analysis. The observed increase in islet graft revascularization and function suggests that our method of stem cell preconditioning may represent a novel strategy to remarkably improve the efficacy of islets-hMSCs cotransplantation.

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Jin, Min. "Activation of Ca²⁺-activated K⁺ Channels and Cell Migration by Hepatocyte Growth Factor/Scatter Factor in Madin-Darby Canine Kidney Cells." [Johnson City, Tenn. : East Tennessee State University], 2002. http://etd-submit.etsu.edu/etd/theses/available/etd-0903102-100659/unrestricted/JinM091302a.pdf.

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25

Pedrazzi, Manuela <1981&gt. "Pathogenesis of cell toxicity by plant toxins." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2013. http://amsdottorato.unibo.it/5247/.

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Ribosome inactivating proteins (RIPs) are a family of plant proteins that depurinate the major rRNA, inhibiting the protein synthesis. RIPs are divided into type 1, single chain proteins with enzymatic activity, and type 2 RIPs (toxic and non-toxic), with the enzymatic chain linked to a binding chain. RIPs have been used alone or as toxic component of immunotoxins for experimental therapy of many diseases. The knowledge of cell death pathway(s) induced by RIPs could be useful for clarifying the mechanisms induced by RIPs and for designing specific immunotherapy. The topic of the current study was (i) the determination of the amino acid sequence of the type 2 RIP stenodactylin. The comparison with other RIPs showed that the A chain is related to other toxic type 2 RIPs. whereas the B chain is more related to the non-toxic type 2 RIPs. This latter result is surprising because stenodactylin is actually the most toxic type 2 RIP known; (ii) the study of the cell death mechanisms induced by stenodactylin in human neuroblastoma cells (NB100). High doses of stenodactylin can activate the effector caspases (perhaps through the DNA damage and/or intrinsic/extrinsic pathways) and also cause ROS generation. Low doses cause a caspase-dependent apoptosis, mainly via extrinsic pathway. Moreover, the activation of caspases precedes the inhibition of protein synthesis; (iii) the investigation of the cell death pathway induced by the non-toxic type 2 RIPs ebulin l and nigrin b. These RIPs demonstrated high enzymatic activity in a cell-free system, but they lack high cytotoxicity. These preliminary studies demonstrate that the cell death mechanism induced by the two non-toxic RIPs is partially caspase-dependent apoptosis, but other mechanisms seem to be involved

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26

Richmond, Sarah Jane. "A study of in situ outer hair cells from the adult mammalian cochlea." Thesis, University of London, 1999. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.313691.

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27

Gould, Fiona Kathryn. "Cloning a novel MAL protein from pancreatic Î²-cells." Thesis, University of Aberdeen, 2004. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.430381.

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Asmall gene family may be involved in sorting proteins to rafts and hence transport to intracellular organelles and the plasma membrane. MAL is found to localise to lipid rafts and is involved in the sorting of apical proteins in polarised epithelial cell types. BENE and Plasmolipin have also been associated with lipid rafts in either endothelial or glial cells. However, MAL, BENE and Plasmolipin are not expressed in endocrine tissues. Recently, a human EST of partial sequence was proposed to be the fourth member of this family. During the current study, the complete coding sequence of this member, termed MAL2, was identified in rat b-cells of the pancreas. Furthermore, through library screening and 5’ RACE, two isoforms of MAL2 were identified - rMAL2-A and rMAL2-B. The two isoforms differ at their amino termini and rMAL2-B contains a long 5’ UTR with upstream AUG (uAUG) codons and two open reading frames. Analysis of these uAUG codons found they were capable of decreasing expression of a reporter gene construct and likely to contribute to the observed low expression of rMAL2-B. Both isoforms were found to be glycosylated on the second extracellular/intralumenal loop. rMAL2-A and rMAL2-B appeared not to display raft association within b-cells, although further work is required to refine the extraction procedure. Subcellular analysis of the two isoforms found that they mainly localised to the Golgi, overlapping in distribution with TGN38, an endogenous TGN marker.

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Parisi, Candida <1979&gt. "Bone-implant interface. Evaluation of osteoblastic cells behavior on nanopatterned titanium surfaces: an in vitro analysis." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2015. http://amsdottorato.unibo.it/7017/.

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Protein-adsorption occurs immediately following implantation of biomaterials. It is unknown at which extent protein-adsorption impacts the cellular events at bone-implant interface. To investigate this question, we compared the in-vitro outcome of osteoblastic cells grown onto titanium substrates and glass as control, by modulating the exposure to serum-derived proteins. Substrates consisted of 1) polished titanium disks; 2) polished disks nanotextured with H2SO4/H2O2; 3) glass. In the pre-adsorption phase, substrates were treated for 1h with αMEM alone (M-noFBS) or supplemented with 10%-foetal-bovine-serum (M-FBS). MC3T3-osteoblastic-cells were cultured on the pre-treated substrates for 3h and 24h, in M-noFBS and M-FBS. Subsequently, the culture medium was replaced with M-FBS and cultures maintained for 3 and 7days. Cell-number was evaluated by: Alamar-Blue and MTT assay. Mitotic- and osteogenic-activities were evaluated through fluorescence-optical-microscope by immunolabeling for Ki-67 nuclear-protein and Osteopontin. Cellular morphology was evaluated by SEM-imaging. Data were statistically analyzed using ANOVA-test, (p<0.05). At day3 and day7, the presence or absence of serum-derived proteins during the pre-adsorption phase had not significant effect on cell-number. Only the absence of FBS during 24h of culture significantly affected cell-number (p<0.0001). Titanium surfaces performed better than glass, (p<0.01). The growth rate of cells between day3 and 7 was not affected by the initial absence of FBS. Immunolabeling for Ki-67 and Osteopontin showed that the mitotic- and osteogenic- activity were ongoing at 72h. SEM-analysis revealed that the absence of FBS had no major influence on cell-shape. • Physico-chemical interactions without mediation by proteins are sufficient to sustain the initial phase of culture and guide osteogenic-cells toward differentiation. • The challenge is avoiding adsorption of ‘undesirables’ molecules that negatively impact on the cueing cells receive from surface. This may not be a problem in healthy patients, but may have an important role in medically-compromised-individuals in whom the composition of tissue-fluids is altered.

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Aamodt, Tor Ingve. "Characterization of ZnS:Cr films for Intermediate Band Solar Cells." Thesis, Norges teknisk-naturvitenskapelige universitet, Institutt for fysikk, 2011. http://urn.kb.se/resolve?urn=urn:nbn:no:ntnu:diva-12828.

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In this thesis samples of ZnS:Cr thin films have been made through physicalvapor deposition (PVD) with resistive heating and characterized by severalmethods. The thin films are of various thicknesses and contain differentconcentration of Cr. ZnS:Cr is a candidate to a bulk material for intermediateband solar cell, IBSC, and was therefore selected for this thesis. IBSC is aconcept for solar cells with a much higher effiency than todays common solarcell.The goal for the project was to put together the deposition system, make thesamples, measure the thicknesses and concentrations and characterize them.Methods like X-Ray Diffraction (XRD), Auger electron spectroscopy (AES),profilometer, four point measurement, Hall-measurement and optical transmissionmeasurements have been used to determine structure, concentrations,thicknesses, resistivity, number of carriers and transmission respectively.The samples and the characterization performed with XRD, AES, profilometer,Hall-measurement and four point measurement were done at NTNU. Thespectrum measurements were done at NTNU and the University in Vienna.Samples with thicknesses in the range of 200 Å to 3340 Å have been investigated.Some were deposited in room temperature, some were annealedafterwards for 2 hours at 4500C and some were deposited on substrates witha temperature of 3000C. The thicknesses were measured with a contact profilometer.The theoretical concentration of the samples were in the range of4% to 31%. There was impossible to measure samples with a lower concentrationthan 7%, even though these were made.Transmission spectra were made in five overlapping specrophotometerss. Twowere used in the visible region (UV-VIS), two in the near infra-red (NIR)region, and one Fourier Transform Infra-Red spectroscopy (FTIR) for thelong wavelength region. The range was from 250 nm up to 25000 nm. NoCr2+ absorption lines were found. This can indicate a cluster form of the Crand not a substitution of Cr2+ with Zn2+ ions. The band gap found fromthe transmission spectra were in the range of >4.5 eV to 2.95 eV. The highband gap imply insulating samples and inactive Cr states in cluster form.No peaks from lattice configuration were found in the XRD spectrum. This indicates an amorphous structure for both the samples deposited in roomtemperature, the annealed samples and the sample deposited on hot substrate.The lattice configuration of the sapphire substrate was found for 3samples. This was the [300] reflection for sapphire at 68.2 degrees.Four samples were investigated in AES. A 3340 Å 7% sample, a 800 Å <5%sample, a 370 Å 31% sample and a 830 Å >4% deposited on hot substrate.Less Cr than expected was found for the sample of 800 Å and 830 Å. However,all samples contained O and C. The sample deposited on hot substrate shouldcontain 80 Å Cr theoretically, but no traces were found. The same appliesfor the 800 Å sample with less than 5%. Both charged badly which implynon-conductive samples. The 31% and the 7% sample did not show the samecharging which implies conductivity. The samples showed a great amount ofCr, but did also contain O and C. However, from the Hall-measurement andfour probe measurement none of the samples showed conductive properties.This emphasize the suspicion of incorporation of Cr clusters in inactive statesmaking the samples insulating.

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Laterza, Claudio <1980&gt. "Circulating Endothelial Progenitor Cells: isolation and biological characterization of EPCs from healthy subjects and nephropatic patients." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2012. http://amsdottorato.unibo.it/4745/.

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Nel 1997 venne isolata una popolazione cellulare con caratteristiche appartenenti a cellule endoteliali mature e a cellule progenitrici ; le cellule appartenenti a queste popolazione furono denominate EPCs (cellule endoteliali progenitrici circolanti) e fu messa in evidenza la loro capacità di dare origine a vasculogenesi postnatale. Lo scopo dello studio è stata la caratterizzazione di tale popolazione cellulare in termini biologici e la valutazione delle differenze delle EPCs in soggetti sani e nefropatici in emodialisi. È stata infine valutata l’eventuale capacità della Vitamina D di influenzare le capacità delle Late EPCs in termini di formazione di colonie in vitro e di attività anticalcifica in soggetti in insufficienza renale cronica.

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31

Guadagnuolo, Viviana <1982&gt. "SMO inhibitor specifically targets the Hedgehog Pathway and reverts the drug-resistance of Leukemic Stem Cells." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2013. http://amsdottorato.unibo.it/5726/.

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Abnormal Hedgehog signaling is associated with human malignancies. Smo, a key player of that signaling, is the most suitable target to inhibit this pathway. To this aim several molecules, antagonists of Smo, have been synthesized, and some of them have started the phase I in clinical trials. Our hospital participated to one of these studies which investigated the oral administration of a new selective inhibitor of Smo (SMOi). To evaluate ex vivo SMOi efficacy and to identify new potential clinical biomarkers of responsiveness, we separated bone marrow CD34+ cells from 5 acute myeloid leukemia (AML), 1 myelofibrosis (MF), 2 blastic phases chronic myeloid leukemia (CML) patients treated with SMOi by immunomagnetic separation, and we analysed their gene expression profile using Affimetrix HG-U133 Plus 2.0 platform. This analysis, showed differential expression after 28 days start of therapy (p-value ≤ 0.05) of 1,197 genes in CML patients and 589 genes in AML patients. This differential expression is related to Hedgehog pathway with a p-value = 0.003 in CML patients and with a p-value = 0.0002 in AML patients, suggesting that SMOi targets specifically this pathway. Among the genes differentially expressed we observed strong up-regulation of Gas1 and Kif27 genes, which may work as biomarkers of responsiveness of SMOi treatment in CML CD34+ cells whereas Hedgehog target genes (such as Smo, Gli1, Gli2, Gli3), Bcl2 and Abca2 were down-regulated, in both AML and CML CD34+ cells. It has been reported that Bcl-2 expression could be correlated with cancer therapy resistance and that Hedgehog signaling modulate ATP-binding (ABC) cassette transporters, whose expression has been correlated with chemoresistance. Moreover we confirmed that in vitro SMOi treatment targets Hedgehog pathway, down-regulate ABC transporters, Abcg2 and Abcb1 genes, and in combination with tyrosine kinase inhibitors (TKIs) could revert the chemoresistance mechanism in K562 TKIs-resistant cell line.

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Ciavarella, Carmen <1986&gt. "Contribution of vascular resident mesenchymal stromal cells to abdominal aortic aneurysm pathogenesis: increased MMP-9 expression and ineffective immunomodulation." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2015. http://amsdottorato.unibo.it/7152/.

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Background. Ageing and inflammation are critical for the occurrence of aortic diseases. Extensive inflammatory infiltrate and excessive ECM proteloysis, mediated by MMPs, are typical features of abdominal aortic aneurysm (AAA). Mesenchymal Stromal Cells (MSCs) have been detected within the vascular wall and represent attractive candidates for regenerative medicine, in virtue of mesodermal lineage differentiation and immunomodulatory activity. Meanwhile, many works have underlined an impaired MSC behaviour under pathological conditions. This study was aimed to define a potential role of vascular MSCs to AAA development. Methods. Aortic tissues were collected from AAA patients and healthy donors. Our analysis was organized on three levels: 1) histology of AAA wall; 2) detection of MSCs and evaluation of MMP-9 expression on AAA tissue; 3) MSC isolation from AAA wall and characterization for mesenchymal/stemness markers, MMP-2, MMP-9, TIMP-1, TIMP-2 and EMMPRIN. AAA-MSCs were tested for immunomodulation, when cultured together with activated peripheral blood mononuclear cells (PBMCs). In addition, a co-colture of both healthy and AAA MSCs was assessed and afterwards MMP-2/9 mRNA levels were analyzed. Results. AAA-MSCs showed basic mesenchymal properties: fibroblastic shape, MSC antigens, stemness genes. MMP-9 mRNA, protein and enzymatic activity were significantly increased in AAA-MSCs. Moreover, AAA-MSCs displayed a weak immunosuppressive activity, as shown by PBMC ongoing along cell cycle. MMP-9 was shown to be modulated at the transcriptional level through the direct contact as well as the paracrine action of healthy MSCs. Discussion. Vascular injury did not affect the MSC basic phenotype, but altered their function, a increased MMP-9 expression and ineffective immunmodulation. These data suggest that vascular MSCs can contribute to aortic disease. In this view, the study of key processes to restore MSC immunomodulation could be relevant to find a pharmacological approach for monitoring the aneurysm progression.

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Bertoni, Sara <1982&gt. "The balance between rRNA and ribosomal protein synthesis up-and down- regulates the tumour suppressor p53 in mammalian cells." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2011. http://amsdottorato.unibo.it/3378/.

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34

Tse, Laam Angela. "Membrane Electrode Assembly (MEA) Design for Power Density Enhancement of Direct Methanol Fuel Cells (DMFCs)." Diss., Georgia Institute of Technology, 2006. http://hdl.handle.net/1853/11522.

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Micro-direct methanol fuel cells (micro-DMFC) can be the power supply solution for the next generation of handheld devices. The applications of the micro-DMFCs require them to have high compactness, high performance, light weight, and long life. The major goal of this research project is to enhance the volumetric power density of direct methanol fuel cells (DMFCs). A performance roadmap has been formulated and showed that patterning the planar membrane electrode assembly (MEA) to 2-D and 3-D corrugated manifolds can greatly increase the power generation with very modest overall volume increases. In this project, different manufacturing processes for patterning MEAs with corrugations have been investigated. A folding process was selected to form 2D triangular corrugations on MEAs for experimental validations of the performance prediction. The experimental results show that the volumetric power densities of the corrugated MEAs have improved by about 25% compared to the planar MEAs, which is lower than the expected performance enhancement. ABAQUS software was used to simulate the manufacturing process and identify the causes of deformations during manufacture. Experimental analysis methods like impedance analysis and 4 point-probes were used to quantify the performance loss and microstructure alteration during the forming process. A model was proposed to relate the expected performance of corrugated MEAs to manufacturing process variables. Finally, different stacking configurations and issues related to cell stacking for corrugated MEAs are also investigated.

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35

Martello, Marina <1983&gt. "The search for Multiple Myeloma Stem cells: molecular characterization and self-renewal mechanisms involved in the disease persistence." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2014. http://amsdottorato.unibo.it/6199/.

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The existence of Multiple Myeloma Stem cells (MMSCs)is supposed to be one of the major causes of MM drug-resistance. However, very little is known about the molecular characteristics of MMSCs, even if some studies suggested that these cells resembles the memory B cells. In order to molecularly characterize MMSCs, we isolated the 138+138- population. For each cell fraction we performed a VDJ rearrangement analysis. The complete set of aberrations were performed by SNP Array 6.0 and HG-U133 Plus 2.0 microarray analyses (Affymetrix). The VDJ rearrangement analyses confirmed the clonal relationship between the 138+ clone and the immature clone. Both BM and PBL 138+ clones showed exactly the same genomic macroalterations. In the BM and PBL 138-19+27+ cell fractions several micro-alterations (range: 1-350 Kb) unique of the memory B cells clone were highlighted. Any micro-alterations detected were located out of any genomic variants region and are presumably associated to the MM pathogenesis, as confirmed by the presence of KRAS, WWOX and XIAP genes among the amplified regions. To get insight into the biology of the clonotypic B cell population, we compared the gene expression profile of 8 MM B cells samples 5 donor B cells vs, thus showing a differential expression of 11480 probes (p-value: <0,05). Among the self-renewal mechanisms, we observed the down-regulation of Hedgehog pathway and the iperactivation of Notch and Wnt signaling. Moreover, these immature cells showed a particular phenotype correlated to resistance to proteasome inhibitors (IRE1α-XBP1: -18.0; -19.96. P<0,05). Data suggested that the MM 138+ clone might resume the end of the complex process of myelomagenesis, whereas the memory B cells have some intriguing micro-alterations and a specific transcriptional program, supporting the idea that these post germinal center cells might be involved in the transforming event that originate and sustain the neoplastic clone.

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Zhang, Xiao. "Preparation and characterization of proton exchange membranes for direct methanol fuel cells." Doctoral thesis, Universitat Rovira i Virgili, 2005. http://hdl.handle.net/10803/8525.

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Due to the petroleum crisis and its consequent emission problems, fuel cells gain an important place in the application of alternative energy. They are a kind of electrochemical device that converts chemical energy directly into electrical energy. The Direct Methanol Fuel Cells (DMFC) use polymer membranes as the electrolyte; the polymer membranes are capable of conducting hydrogen protons. The fuel cell system is still expensive and the proton exchange membrane has contributed significantly the high cost. At present, perfluorosulfonic acid membranes (PFSA) (e.g. Nafion®, by DuPont) have been widely investigated. However they showed high methanol crossover and high swelling that lead low cell efficiency.
The main goal of the thesis is to prepare novel proton exchange membranes to apply in the DMFC. PEG and PA membranes compuestas fueron preparadas. Derivados del ácido fosfórico y lignosulfonados (LS) fueron incluidos en la estructura de la PA para actuar como agentes transportadores de protones. El mecanismo de la conductividad de protón es "hopping". Ellos mostraron el más baja del transporte de metanol.
Se obtuvieron también membranas híbridas de LS, preparadas mediante la mezcla de los dos polímeros, LS y PSU, siguiendo el método de precipitación en inmersión. Las propiedades electroquímicas de las membranas de LS fueron caracterizadas. Las membranas de LS alcanzaron conductividades de protón aceptables (10-20 mS/cm) con capacidad de intercambio iónico muy baja (IEC) (60 veces más baja que Nafion). "Membrane electrode assemblies" (MEAs) fueron preparadas y sus rendimientos de celda fueron medidos en una celda individual directa de metanol (DMFC).
LS membrana is the highlight point of this thesis. It demonstrated the first that LS is a good proton exchange material although it is a waste from the paper industry. It also proved that porous membrane can be used in the DMFC with acceptable proton conductivity and low methanol permeability, which is a totally new way from the existing literatures.
The results have been published on international journals and have been presented on international conferences:

1. X. Zhang, A. Glüsen, R. Garcia-Valls, Porous Lignosulfonate membrane for direct methanol fuel cells, accepted by Journal of Membrane Science, 2005
2. X. Zhang, J. Benavente, R. Garcia Valls, Lignin-based Membranes for Electrolyte Transference, Journal of Power Sources, 145 (2005) 292
3. X. Zhang, L. Pitol Filho, C. Torras, R. Garcia Valls, Experimental and Computational Study of Proton and Methanol Permeability through Composite Membranes, Journal of Power Sources, 145 (2005) 223
4. J. Benavente, X. Zhang, R. Garcia Valls, Modification of Polysulfone Membranes with Polyethylene Glycol and Lignosulfate: Electrical Characterization by Impedance Spectroscopy Measurements, Journal of Colloid and Interface Science, 285 (2005) 273-280
5. X. Zhang, R. Garcia-Valls, Proton transport membrane containing lignin compound for direct methanol fuel cells (Poster), 5th Ibero American Congress on Membrane Science and Technology, 2005, Valencia- Spain
6. X. Zhang, J. Benavente and R. Garcia-Valls, Lignin-based membranes for electrolyte transference (Oral presentation), Fuel Cell Science & Technology, Oct. 2004, Munich- Germany.
7. X. Zhang, R. Garcia-Valls, New membranes for Proton Transport in DMFC (Poster), Euromembrane Sep. 2004, ISBN: 3-930400-65-0, p. 64, Hamburg- Germany,
8. X. Zhang, R. Garcia-Valls, Lignosulfonate Application in Proton Transport Membrane (Oral presentation), 2nd World Conference and Technology Exhibition on Biomass for Energy, Industry and Climate Protection, May. 2004, Rome- Italy
9. X. Zhang, R. Garcia-Valls, Proton Selective Composite Membrane for Direct Methanol Fuel Cell (Oral presentation), 5th NYM (Network Young Membrains) Oct. 2003, ISBN: 84-688-3132-8, p. 199, Barcelona, Spain
10. X. Zhang, R. Garcia-Valls, A. Jiménez-López, E. Rodríguez-Castellón and J. Benavente, Electrical and Chemical Surface Characterization of Lignosulfate/Polysulfone Membranes for Fuel Cells Application, International Conference on "New Proton Conducting Membranes and Electrodes for PEM FCs", Oct. 2005, Assisi, Italy.
Debido a la crisis de petróleo y a los problemas de emisión, las pilas de combustible adquieren un lugar importante en la aplicación de la energía alternativa. Son una clase de dispositivo electroquímico que convierte la energía química directamente en energía eléctrica. Las celdas de combustible de metanol (DMFC) usan membranas de polímero como el electrolito; las membranas de polímero son capaces de transportar protones de hidrógeno. El sistema de la celda de combustible todavía es costoso y las membranas de intercambio de protón han contribuido significativamente para el costo elevado.
Actualmente, las membranas de ácido perfluorosulfonico (PFSA) (por ejemplo, Nafion ®, de DuPont) ten sido investigadas extensamente. Sin embargo mostraron alto paso de metanol e alto "swelling" lo que lleva a una eficiencia de celda baja.
El objetivo principal de la tesis es preparar membranas de intercambio de protón nuevas para la aplicación en DMFC. Membranas compuestas de PEG y de PA fueron preparadas. Derivados del ácido fosfórico y lignosulfonados (LS) fueron incluidos en la estructura de la PA para actuar como agentes transportadores de protones. El mecanismo de conductividad de protón es "hopping". Ellos mostraron el transporte de metanol más bajo.
Se obtuvieron también membranas híbridas de LS, preparadas mediante la mezcla de los dos polímeros, LS y PSU, siguiendo el método de precipitación en inmersión. Las propiedades electroquímicas de las membranas de LS fueron determinadas. Las membranas de LS alcanzaron conductividades de protón aceptables (10-20 mS/cm) con capacidad de intercambio iónico muy baja (IEC) (60 veces más baja que Nafion). "Membrane electrode assemblies" (MEAs) fueron preparadas y sus rendimientos de celda fueron medidos en una celda individual directa de metanol (DMFC).
Las membranas de LS son el punto principal de esta tesis. Primero se demostró que LS es un material de intercambio de protón muy bueno aunque sea un residuo de la industria de papel. También se probó que membranas porosas pueden ser usadas en DMFC con una conductancia de protón aceptable y baja permeabilidad de metanol, lo que es una manera totalmente nueva comparada a la literatura existente.
Los resultados han sido divulgados en revistas internacionales y han sido presentados en conferencias internacionales:
1. X. Zhang, A. Glüsen, R. Garcia-Valls, Porous Lignosulfonate membrane for direct methanol fuel cells, accepted by Journal of Membrane Science, 2005
2. X. Zhang, J. Benavente, R. Garcia Valls, Lignin-based Membranes for Electrolyte Transference, Journal of Power Sources, 145 (2005) 292
3. X. Zhang, L. Pitol Filho, C. Torras, R. Garcia Valls, Experimental and Computational Study of Proton and Methanol Permeability through Composite Membranes, Journal of Power Sources, 145 (2005) 223
4. J. Benavente, X. Zhang, R. Garcia Valls, Modification of Polysulfone Membranes with Polyethylene Glycol and Lignosulfate: Electrical Characterization by Impedance Spectroscopy Measurements, Journal of Colloid and Interface Science, 285 (2005) 273-280
5. X. Zhang, R. Garcia-Valls, Proton transport membrane containing lignin compound for direct methanol fuel cells (Poster), 5th Ibero American Congress on Membrane Science and Technology, 2005, Valencia- Spain
6. X. Zhang, J. Benavente and R. Garcia-Valls, Lignin-based membranes for electrolyte transference (Oral presentation), Fuel Cell Science & Technology, Oct. 2004, Munich- Germany.
7. X. Zhang, R. Garcia-Valls, New membranes for Proton Transport in DMFC (Poster), Euromembrane Sep. 2004, ISBN: 3-930400-65-0, p. 64, Hamburg- Germany,
8. X. Zhang, R. Garcia-Valls, Lignosulfonate Application in Proton Transport Membrane (Oral presentation), 2nd World Conference and Technology Exhibition on Biomass for Energy, Industry and Climate Protection, May. 2004, Rome- Italy
9. X. Zhang, R. Garcia-Valls, Proton Selective Composite Membrane for Direct Methanol Fuel Cell (Oral presentation), 5th NYM (Network Young Membrains) Oct. 2003, ISBN: 84-688-3132-8, p. 199, Barcelona, Spain
10. X. Zhang, R. Garcia-Valls, A. Jiménez-López, E. Rodríguez-Castellón and J. Benavente, Electrical and Chemical Surface Characterization of Lignosulfate/Polysulfone Membranes for Fuel Cells Application, International Conference on "New Proton Conducting Membranes and Electrodes for PEM FCs", Oct. 2005, Assisi, Italy La tesis tuvo la cooperación del Forschungszentrum Jülich, Alemania y la doctoranda esta solicitando el titulo de Doctorado Europeo.

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Wallace, Jamie Stuart. "Development of a Carbon Dioxide Continuous Scrubber (CDOCS) System for Alkaline Fuel Cells." Thesis, University of Canterbury. Mechanical Engineering, 2006. http://hdl.handle.net/10092/1077.

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Alkaline fuel cells (AFC's) using renewable fuels are a developing technology capable of meeting market niches in standby, standalone and distributed power generation. AFC's generate electricity, heat and water using hydrogen and oxygen as fuels. While AFC's have been known and the principles demonstrated for over sixty years, their use has been restricted primarily to space applications. Recent technological developments have seen the cost of AFC stacks fall considerably; this together with several other advantages over competing fuel cell technology, has rekindled interest in commercial systems. The main deterrent to wide spread commercialisation of AFC systems is susceptibility to carbon dioxide (CO2) in atmospheric air used as the oxygen supply. AFC's require a low cost, low energy, continuous scrubbing device to reduce CO2 in air from approximately 380 parts per million (ppm) atmospheric concentration to below 50 ppm. Current technology to overcome this problem, a solid expendable absorbent called soda lime, is not viable for commercial systems. The project scope included concept generation of a device to remove CO2 from air, the development of a CO2 measurement technique, investigation of chemistry and flow phenomena to determine design relations, and product design and embodiment. The scrubber system conceived specifically for AFC systems uses the temperature swing chemistry of a liquid chemical absorbent, monoethanolamine, and a packed bubble column apparatus to provide intimate gas-liquid interaction. Prototype development proved the Carbon Dioxide Continuous Scrubber (CDOCS) concept and a Patent Cooperation Treaty (PCT) patent was granted, followed by a full American patent. A gas chromatographic measurement technique was developed to measure low ppm concentration CO2 in air, enabling regular monitoring of scrubbed gas. Carbon dioxide was separated from a small sample of scrubbed air by chromatographic columns, and the gases analysed with a thermal conductivity detector. The GC system was capable of measuring to 10 ppm with good resolution and accuracy. Experimental studies were carried out to characterise the flow dynamics and absorption phenomena in the packed bubble column absorber. The relationship between absorption performance and gas-liquid contact time, an important operating parameter for use with AFC's, was theoretically determined and later confirmed by experiment. The regeneration process was studied and the optimal regenerator design determined to be second, smaller packed bubble column. Experiments were conducted to establish design relations for regeneration temperature, flush gas flow rate and the effect of multiple regeneration cycles. A prototype CDOCS system was built to enable experimental characterisation of scrubbing performance as a function of primary design and operating parameters including liquid depth, regenerator operating temperature and solution composition. This resulted in a good understanding of the system, and an optimised experimental run was performed for cost and performance comparison to existing scrubbing technology. The CDOCS was capable of reducing CO2 in air from 380 to 80 ppm for thirty days, providing low cost, low maintenance scrubbing compared to soda lime. The capital cost of the CDOCS is considerably more than for soda lime scrubbers, and the penalty for extended operation is parasitic power consumption by the CDOCS system totalling less than 7% of fuel cell output. It is suggested that a combination of the two technologies be used initially to provide effective, low cost scrubbing for AFC and CDOCS co-development. Future work on the CDOCS project should include reduction of chemical vapour carry over to the fuel cell, followed by integration with an AFC system. This would allow further development, refinement and design for production to reduce capital cost.

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Brighenti, Elisa <1981&gt. "Relevance of cell cycle regulators on chemotherapy response in breast cancer." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2011. http://amsdottorato.unibo.it/3454/.

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39

Timme, Cindy R. "Drug Resistance Mechanisms to Gamma-secretase Inhibitors in Human Colon Cancer Cells." Scholar Commons, 2013. http://scholarcommons.usf.edu/etd/4954.

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Colorectal cancer is the third leading cause of cancer-related mortality. Much progress has been achieved in combating this disease with surgical resection and chemotherapy in combination with targeted drugs. However, most metastatic patients develop drug resistance so new modalities of treatment are needed. Notch signaling plays a vital role in intestinal homeostasis, self-renewal, and cell fate decisions during post-development and is activated in colorectal adenocarcinomas. Under debate is its role in carcinomas and metastatic disease. In theory, blocking Notch activation using gamma-secretase inhibitors (GSIs) may show efficacy alone or in combination with chemotherapy in the treatment of colon cancer. In Chapter Three, we tested the capacity for GSIs to synergize with oxaliplatin in colon cancer cell lines and evaluated the underlying molecular mechanisms. GSI alone had no effect on colon cancer cell lines. Surprisingly, we show that GSIs blocked oxaliplatin-induced apoptosis through increased protein levels of the anti-apoptotic Bcl-2 proteins Mcl-1 and/or Bcl-xL. Restoration of apoptosis was achieved by blocking Mcl-1 and/or Bcl-xL with obatoclax (an anti-apoptotic Bcl-2 agonist) or siRNA. An unexpected result was the induction of cell death with the combination of GSI and obatoclax. In Chapter Four, we examined the mechanism of GSI + obatoclax-mediated cell death. We found that apoptosis played a minimal role. Rather, we identified blockage of cytoprotective autophagy played a causative role. Interestingly, we also saw autophagy induction in GSI-treated cells, which could explain the insensitivity of colon cancer cells to GSI. When autophagy was blocked in GSI-treated cells, cells became sensitive to GSI and cell death was elicited. The mechanism by which induction of autophagy occurs in GSI- treated cells is an area for further research. Overall, our work questions the validity of the use of GSIs in the treatment of colorectal cancers. We show that GSIs may block apoptosis and induce cytoprotective autophagy simultaneously, resulting in increased drug resistance and cellular survival. Whether these two cellular survival processes occurs in patients needs to be examined before GSIs can be utilized in a clinical setting. If so, these two hurdles must be overcome.

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40

Onofrillo, Carmine <1984&gt. "Ribosome Biogenesis and cell cycle regulation: Effect of RNA Polymerase III inhibition." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2013. http://amsdottorato.unibo.it/5422/.

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In cycling cells positive stimuli like nutrient, growth factors and mitogens increase ribosome biogenesis rate and protein synthesis to ensure both growth and proliferation. In contrast, under stress situation, proliferating cells negatively modulate ribosome production to reduce protein synthesis and block cell cycle progression. The main strategy used by cycling cell to coordinate cell proliferation and ribosome biogenesis is to share regulatory elements, which participate directly in ribosome production and in cell cycle regulation. In fact, there is evidence that stimulation or inhibition of cell proliferation exerts direct effect on activity of the RNA polymerases controlling the ribosome biogenesis, while several alterations in normal ribosome biogenesis cause changes of the expression and the activity of the tumor suppressor p53, the main effector of cell cycle progression inhibition. The available data on the cross-talk between ribosome biogenesis and cell proliferation have been until now obtained in experimental model in which changes in ribosome biogenesis were obtained either by reducing the activity of the RNA polymerase I or by down-regulating the expression of the ribosomal proteins. The molecular pathways involved in the relationship between the effect of the inhibition of RNA polymerase III (Pol III) activity and cell cycle progression have been not yet investigated. In eukaryotes, RNA Polymerase III is responsible for transcription of factors involved both in ribosome assembly (5S rRNA) and rRNA processing (RNAse P and MRP).Thus, the aim of this study is characterize the effects of the down-regulation of RNA Polymerase III activity, or the specific depletion of 5S rRNA. The results that will be obtained might lead to a deeper understanding of the molecular pathway that controls the coordination between ribosome biogenesis and cell cycle, and might give useful information about the possibility to target RNA Polymerase III for cancer treatment.

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D’Anello, Laura <1980&gt. "Epigenetic control of the basal-like gene expression profile via Interleukin-6 in breast cancer cells." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2011. http://amsdottorato.unibo.it/3368/.

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42

Chatgilialoglu, Alexandros <1981&gt. "Membrane lipidomics: the reorganization of fatty acids as a biomarker of cell condition." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2009. http://amsdottorato.unibo.it/2145/.

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43

Julião, Paulo Sérgio Barros. "Electrolytic cells for plastic waste recycling." Master's thesis, Universidade de Aveiro, 2014. http://hdl.handle.net/10773/15804.

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Mestrado em Sistemas Energéticos Sustentáveis
The current project assesses potential molten alloy anodes for Solid Oxide Fuel Cells (SOFC) running on solid waste. A detailed phase diagram study was performed to locate probable anode systems. The molten metal oxide system PbO-Sb2O3 was selected as a possible molten alloy anode for this application. A detailed vapour pressure study of this system was performed. Several cells were fabricated to experimentally assess the electrochemical properties of this system. The work reveals several unexpected limiting features such as the incompatibility between the platinum and the chosen alloy. A second cell was built, this time using rhenium wires instead, preventing such reaction. However, the rhenium wire sublimes under oxidizing conditions (air) and the sealing glass and the chosen alloy system react with each other under long term use. Considering all these issues, a third cell design was conceived, surpassing some obstacles and providing some initial information regarding the electrochemical behaviour. The current project shows that many parameters need to be taken into account to ensure materials compatibility. For the PbOSb2O3 system, the high volatility of Sb2O3 was a serious limitation that can only be addressed through the application of new contact wires or sealing materials and conditions. Nonetheless, the project highlights several other potential systems that can be considered, such as Pb11Ge3O17, Pb3GeO5, Pb5Ge3O11, Bi2CuO4, Bi2PdO4, Bi12GeO20.
Este estudo incidiu sobre potenciais ânodos líquidos de ligas metálicas para células electrolíticas (do tipo SOFC) alimentadas por resíduos sólidos. Alguns sistemas de ânodos possíveis foram identificados através de um estudo detalhado de diagramas de fase. O sistema de óxidos metálicos PbO-Sb2O3 foi selecionado como uma possível liga metálica para esta aplicação. Este sistema foi sujeito a um estudo detalhado de pressão de vapor. Algumas células foram fabricadas para avaliar experimentalmente as propriedades electro-químicas deste sistema. Este trabalho revela imensas características que inesperadamente limitaram este estudo, tal como a incompatibilidade entre platina e a liga metálica escolhida. Uma segunda célula foi construída, desta vez usando um fio de rénio, prevenindo tal reacção. No entanto, o fio de rénio sublima sobre condições oxidantes (ar) e, perante um uso prolongado, o vidro selante e a liga metálica reagem entre si. Considerando todas estas incompatibilidades, um terceiro modelo de célula foi criado, ultrapassando alguns obstáculos e fornecendo alguma informação inicial relativa ao comportamento electro-químico. O presente trabalho mostra que vários parâmetros precisam precisam de ser abordados de modo a assegurar a compatibilidade dos materiais. Relativamente ao sistema PbO-Sb2O3, a elevada volatilidade de Sb2O3 foi uma grave limitação que só pode ser contornada através da aplicação de novos fios conectores, materiais e condições de selamento. No entanto, este projecto destaca outros potenciais sistemas que podem ser estudados, como Pb11Ge3O17, Pb3GeO5, Pb5Ge3O11, Bi2CuO4, Bi2PdO4, Bi12GeO20.

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Santos, Antonio Rodolfo dos. "Análise por impedância eletroquímica \"on-line\" de conjuntos eletrodo/membrana (MEA) de células a combustível a membrana polimérica (PEMFC)." Universidade de São Paulo, 2007. http://www.teses.usp.br/teses/disponiveis/85/85134/tde-16052012-093213/.

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Este trabalho apresenta resultados de estudos e caracterizações de Conjuntos Eletrodo/Membrana (MEAs) de Células a Combustível a Membrana Polimérica (PEMFC). Algumas condições de operação de células e diferentes processos de produção de MEA foram investigados. A técnica de Espectroscopia de Impedância Eletroquímica (EIE) (em situ - 0 a 16 A) foi usada \"on-line\" como uma ferramenta de diagnóstico, relativa ao desempenho de célula. As medidas de EIE foram feitas através do Sistema de EIE para células a combustível FC350 (GAMRY), junto a um PC4 Potentiostato/Galvanostato e conectado à carga dinâmica (TDI) para experimentos de EIE \"on-line\" (100 mHz - 10 kHz, dU = 5 mV). MEAs com 25 cm2 de área ativa, usando eletrocatalisadores PtM/C 20 % (M = Ru, Sn ou Ni) fabricados usando o Método de Redução por Álcool (MRA). A tinta catalítica foi diretamente aplicada no Tecido de Carbono (GDL) e este prensado na membrana de Nafion® (105). MEAs usando eletrocatalisadores Pt/C e PtRu/C 20 % da E-TEK foram fabricados para comparação. Todos os cátodos foram confeccionados com Pt/C 20% da E-TEK. Foram fixadas as concentrações de metal nobre em 0,4 mg Pt.cm-2 no anodo e 0,6 mg Pt.cm-2 no catodo (E-TEK). Diagramas de Nyquist dos MEAs com Pt/C e PtRu/C da E-TEK ou PtM/C MRA apresentaram as mesmas resistências de ôhmicas para os MEAs. Este fato pode ser explicado por supressão de aglomerados durante o processo de preparação do MEA ou pela homogeneidade do eletrocatalisador ancorado ao carbono. Também pôde ser observado, a baixas densidades atuais que há uma diferença de desempenho significante entre o eletrocatalisadores da ETEK e os preparados pelo MRA. Os resultados das curvas de polarização confirmaram que PtM/C MRA apresentara um aumento de atividade para as células alimentadas com metanol e etanol. A técnica de EIE se mostrou eficiente para a avaliação do método de preparação dos MEAs e do desempenho da célula, os resultados de EIE mostraram uma coerência na escolha do modelo do circuito elétrico para os MEAs utilizando hidrogênio, metanol e etanol. Esta coerência indica que outras resistências não consideradas no modelo não são relevantes na resistência total dos MEAs.
This work reports results of studies and characterization on Membrane Electrode Assemblies (MEAs) for Proton Exchange Membrane Fuel Cell (PEMFC). Some cell operation conditions and different processes of MEA production were investigated. The Electrochemical Impedance Spectroscopy Technique (EIS) (in situ - 0 to 16 A) was used \"on-line\" as a tool for diagnosis, concerning the cell performance. The EIS measurements were carried out with a FC350 Fuel Cell EIS System (GAMRY), coupled to a PC4 Potentiostat/Galvanostat and connected to the electronic load (TDI) for \"on-line\" EIS experiments (100 mHz - 10 kHz, dU = 5 mV). MEAs with 25 cm2 surface area, using PtM/C 20% (M = Ru, Sn or Ni) electrocatalysts were manufactured using the Alcohol Reduction Process (ARP). The catalytic ink was applied directly into the Carbon Cloth (GDL) and pressed in the NafionR membrane (105). MEAs using Pt/C and PtRu/C 20% from E-TEK electrocatalysts were manufactured by comparison. All the cathodes were sprayed with Pt/C 20% from E-TEK. The noble metal concentrations used were set to 0.4 mg Pt.cm-2 at the anode and 0.6 mg Pt.cm-2 at the cathode (E-TEK). Nyquist diagrams of the MEAs with Pt/C and PtRu/C from E-TEK or PtM/C (M = Ru, Sn or Ni) ARP showed essentially the same ohmic resistances for the MEAs. This fact can be explained by suppression of agglomerates during the MEA preparation process or by the homogeneity of the anchored electrocatalysts at the carbon surface. It could also be observed, at low current densities, that there was a significant performance difference between the electrocatalysts from E-TEK and those prepared with the Alcohol Reduction Process. The polarization curves results confirmed that the PtM/C (M = Ru, Sn or Ni) ARP showed an activity increase for the methanol and ethanol fed cells. The technique of EIE was shown efficient for the evaluation of the method preparation of MEAs and the acting of the cell, the results of EIE showed coherence in the choice of the model the electric circuit for MEAs using hydrogen, methanol and ethanol. This coherence indicates that other resistances no considered in the model are not relevant in the total resistance of MEAs.

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Chladil, Ladislav. "Optimalizace MEA struktury pro nízkoteplotní palivové články." Master's thesis, Vysoké učení technické v Brně. Fakulta elektrotechniky a komunikačních technologií, 2010. http://www.nusl.cz/ntk/nusl-218680.

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This master’s thesis focuses on optimization of electrode configuration in combination with a polymer membrane (MEA - Membrane Electrode Assembly) in terms of material and technology. The main goal was to create a functional measurement of MEA structures with three different types of carbon materials. The theoretical part focuses on the physical and chemical properties of low-temperature fuel cells with polymer membrane. The experimental section describes the manufacture of catalytic materials with different types of carbon and various contents platinum. Produced by electrode materials were investigated by cyclic voltammetry. The next step was to manufacture MEA structures and characteristics of VA measurement using a digitally controlled load in the experimental fuel cell Quintech.

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Xiao, Kang. "Investigating the functional roles of Mcl-1 in apoptosis in mammalian cells /." View abstract or full-text, 2009. http://library.ust.hk/cgi/db/thesis.pl?BIOL%202009%20XIAO.

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Pollutri, Daniela <1982&gt. "Drugs down-regulating E2F-1 expression hinders cell proliferation through a p53-indipendent mechanism." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2014. http://amsdottorato.unibo.it/6575/.

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E2F-1 is a transcription factor that plays a key role in cell-cycle control at G1/S check-point level by regulating the timely expression of many target genes whose products are required for S phase entry and progression. In mammalian cells, E2F-1 is negatively regulated by hypo-phosphorylated Retinoblastoma protein (pRb) whereas it is protected against degradation by its binding to Mouse Double Minute 2 protein (MDM2). In this study we experimented a drug combination in order to obtain a strong down-regulation of E2F-1 by acting on two different mechanisms of E2F-1 regulation mentioned above. This was achieved by combining drugs inhibiting the phosphorylation of pRb with drugs inactivating the MDM2 binding capability. The mechanism of action of these drugs in down-regulating E2F-1 level and activity is p53 independent. As expected, when combined, these drugs strongly inhibits E2F-1 and hinder cell proliferation in p53-/- and p53-mutated cells by blocking them in G1 phase of cell cycle, suggesting that E2F-1 down-regulation may represent a valid chemotherapeutic approach to inhibit proliferation in tumors independently of p53 status.

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Sharma, Manu. "Host cell death modulation by Chlamydia trachomatis." Doctoral thesis, Humboldt-Universität zu Berlin, Mathematisch-Naturwissenschaftliche Fakultät I, 2010. http://dx.doi.org/10.18452/16253.

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Chlamydien durch die Modulation spezifischer Wirtszellproteine verschiedene Wege der Apoptose verhindern können. Mcl-1 und cIAP-2 erwiesen sich als bedeutende Faktoren, die durch die Infektion hochreguliert und absolut notwendig für die Inhibierung der Apoptose durch Chlamydien waren. Hochregulation der Mcl-1 Expression führte zu einem Block im apoptotischen Weg oberhalb der Mitochondrien. cIAP-2 zusammen mit anderen Inhibitor of Apoptosis Proteins (IAP) verhinderten die Aktivierung von Caspase-3, denfinalen Schritt in der apoptotischen Kaskade. Weiterhin wurde beobachtet, dass die Aktivierung des MAPKinase-Signalweges durch die Infektion wichtig war für die Hochregulierung von Mcl-1 und cIAP-2. Ein Hochdurchsatz-Screen wurde durchgeführt, um andere Wirtszellfaktoren, die für die Apoptoseinhibierung durch die Chlamydien verantwortlich sind, zu identifizieren. Neben Mcl-1 waren die identifizierten Faktoren hauptsächlich Mitglieder des MAPKinase-Signalweges. Dabei wurde deren Rolle für die Apoptoseresistenz bestätigt. Eine weiterführende Analyse der im Screen ermittelten Faktoren identifizierte eine Funktion von HIF-1a bei der Modulation der Expression anti-apoptotischer Faktoren während der Infektion. Es wurde beobachtet, dass HIF-1a stabilisiert und zum Nukleus transloziert wird. Es ist bekannt, dass HIF-1a HIF-1a im Nukleus binden kann, um den funktionalen Transkriptionsfaktor HIF zu bilden. Dieser reguliert die Expression verschiedener Überlebensfaktoren, unter anderem Mcl-1. HIF-1a Knockdown inhibierte die Chlamydien-induzierte Hochregulation von Mcl-1 mRNA-Expression.
chlamydial infection blocked the apoptotic pathway at multiple levels by modulation of specific host cell proteins. Mcl-1 and cIAP-2 were two most prominent factors that were up-regulated during the infection, and absolutely required for apoptosis inhibition. Increased expression of Mcl-1 led to a block in the apoptotic pathway upstream of the mitochondria. cIAP-2, together with other inhibitor of apoptosis proteins (IAPs), blocked the activation of caspase-3 at the final step of the apoptosis cascade. Further, it was observed that the activation of the MAPK pathways during infection was needed for the up-regulation of Mcl-1 and cIAP-2. A high throughput RNAi screen was performed to identify other host factors required for the apoptosis resistance during the infection. Besides Mcl-1, the targets from the screen prominently included members of the MAPK pathways, confirming their role in the apoptosis resistance. Pathway analysis of the targets identified the role of HIF-1a in modulating the expression of the anti-apoptotic factors during infection. It was observed that during infection, HIF-1a gets stabilized and translocates to the nucleus. It is known that HIF-1a can bind to HIF-1a in the nucleus to form the functional transcription factor HIF, which can regulate the expression of survival factors like Mcl-1. This was seen to be the case, because knock down of HIF-1a abrogated the infection induced up-regulation of Mcl-1 at the mRNA levels.

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Etebari, Maryam <1983&gt. "Toward a Molecular Classification of Peripheral T-Cell Lymphomas: The Role of Gene Expression Profiling." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2016. http://amsdottorato.unibo.it/7434/.

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Peripheral T-cell lymphomas-not otherwise specified (PTCL/NOS) are the most common T-cell neoplasms. This study sought to reshape the PTCL/NOS sub-classification (including its two main morphological variants, Lennert lymphoma, LL, and Follicular variant, F-PTCL) based on the correspondence between their molecular features and those of different functional T-cell subsets, also assessing the clinical impact of such an approach. We found that PTCLs/NOS could be divided into groups corresponding to T-cell subsets differently reliant on transcription regulators including mTOR and FOXP3, and identified minimal gene sets discriminating among these groups. Notably, by grouping tumors according to their dependency on master regulators of T-lymphocyte fate, we identified three groups (T-cytotoxic, Treg/TFH, and other-T-helper) characterized by specific genetic patterns and significantly different clinical outcomes. Immunohistochemistry partially substituted for the molecular analysis by consistently recognizing only Treg and TFH cases. Finally, targeted inhibition of MTOR in T-helper cases (that were characterized by genetic lesions targeting the pathway) was proved to be effective ex vivo. We conclude that PTCL/NOS can be divided into subgroups corresponding to different cellular counterparts, characterized by different genetic patterns and possibly sensitivity to specific therapeutic approaches. Furthermore, we identified different gene and microRNA signatures for LL capable of differentiating it from other PTCL/NOS and enriched in cytotoxic function. Moreover, PI3K/Akt/mTOR pathway emerged as novel therapeutic targets for LL. Additionally, LL showed some differences with other PTCL/NOS in terms of clinical features, all supporting its recognition as a distinct entity. Besides, we found that F-PTCL has a distinct molecular signature more similar to PTCL/NOS rather than AITL, and therefore cannot be included among AITLs at least based on GEP, although this necessities more genetic studies. Overall, these results may impact on PTCL classification as well as on future studies aimed to define the more appropriate therapeutic strategy for each identified subgroup/entity.

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Papayannidis, Cristina <1980&gt. "Pre-clinical and clinical development of leukemia stem cell inhibitors in acute leukemias." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2013. http://amsdottorato.unibo.it/5596/.

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In Leukemias, recent developments have demonstrated that the Hedgehog pathway plays a key-role in the peculiar ability of self renewal of leukemia stem cells. The aim of this research activity was to investigate, through a first in man, Phase I, open label, clinical trial, the role and the impact, mainly in terms of safety profile, adverse events and pharmacokinetics, of a Sonic Hedgehog inhibitor compound on a population of heavely pretreated patients affected by AML, CML, MF, or MDS, resistant or refractory to standard chemotherapy. Thirty-five patients have been enrolled. The drug was administered orally, in 28 days cycles, without rest periods. The compound showed a good safety profile. The half life was of 17-35 hours, justifying the daily administration. Significant signs of activity, in terms of reduction of bone marrow blast cell amount were seen in most of the patients enrolled. Interestingly, correlative biological studies demonstrated that, comparing the gene expression profyiling signature of separated CD34+ cells before and after one cycle of treatment, the most variably expressed genes were involved in the Hh pathway. Moreover, we observed that many genes involved in MDR (multidrug resistance)were significantly down regulated after treatment. These data might lead to future clinical trials based on combinatory approaches, including, for instance, Hh inhibitors and conventional chemotherapy.

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Dissertations / Theses on the topic 'MEL cells'

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