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1
Shen, Yang. "A high-resolution genetic map of human chromosome 16 and localization of the MEF gene /." Title page, contents and summary only, 1994. http://web4.library.adelaide.edu.au/theses/09PH/09phs546.pdf.
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Thesis (Ph. D.)--University of Adelaide, Dept. of Paediatrics, Women's and Children's Hospital, 1994.Copies of author's previously published articles inserted. Includes bibliographical references.
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Accardo, Silvia <1976>. "Il gene plasmidico orf5 e il gene pmpD di Chlamydia trachomatis." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2010. http://amsdottorato.unibo.it/2911/1/ACCARDO_SILVIA_TESI.pdf.
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Accardo, Silvia <1976>. "Il gene plasmidico orf5 e il gene pmpD di Chlamydia trachomatis." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2010. http://amsdottorato.unibo.it/2911/.
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Knigge, Anja, Nora Klöting, Michael R. Schön, Arne Dietrich, Mathias Fasshauer, Daniel Gärtner, Tobias Lohmann, et al. "ADCY5 gene expression in adipose tissue is related to obesity in men and mice." Universitätsbibliothek Leipzig, 2015. http://nbn-resolving.de/urn:nbn:de:bsz:15-qucosa-169954.
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Genome wide association studies revealed an association of the single nucleotide polymorphism rs11708067 within the ADCY5 gene—encoding adenylate cyclase 5—with increased type 2 diabetes (T2D) risk and higher fasting glucose. However, it remains unclear whether the association between ADCY5 variants and glycemic traits may involve adipose tissue (AT) related mechanisms. We therefore tested the hypothesis that ADCY5 mRNA expression in human and mouse AT is related to obesity, fat distribution, T2D in humans and high fat diet (HFD) in mice. We measured ADCY5 mRNA expression in paired samples of visceral and subcutaneous adipose tissue from 244 individuals with a wide range of body weight and parameters of hyperglycemia, which have been genotyped for rs11708067. In addition, AT ADCY5 mRNA was assessed in C57BL/6NTac which underwent a 10 weeks standard
chow (n = 6) or high fat diet (HFD, n = 6). In humans, visceral ADCY5 expression is significantly higher in obese compared to lean individuals. ADCY5 expression correlates with BMI, body fat mass, circulating leptin, fat distribution, waist and hip circumference, but not with fasting plasma glucose and HbA1c. Adcy5 expression in mouse AT is significantly
higher after a HFD compared to chow (p<0.05). Importantly, rs11708067 is not associated with ADCY5 mRNA expression levels in either fat depot in any of the genetic models tested. Our results suggest that changes in AT ADCY5 expression are related to obesity and fat distribution, but not with impaired glucose metabolism and T2D. However, altered ADCY5 expression in AT does not seem to be the mechanism underlying the association between rs11708067 and increased T2D risk.
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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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Tourlaki, Athanasia <1973>. "The KIT gene in familial mastocytosis." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2013. http://amsdottorato.unibo.it/5590/1/Tourlaki_Athanasia_tesi.pdf.
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Familial cutaneous mastocytosis is an exceptional condition of unknown etiology. In this study we report the largest series of patients with familial cutaneous mastocytosis without other manifestations (18 affected subjects from seven unrelated families), and we investigate the role of germ-line KIT mutations in the pathogenesis of the disease. The mean age at onset was 5.4 years (range from birth to 22 years), and the clinical behavior was variable over a mean follow up period of 15.1 years (range 2-36): improvement in seven, stability in eight and worsening in the remaining three patients. The pattern of inheritance was compatible with an autosomal dominant trait with incomplete penetrance; a female preponderance (14 females vs 4 males, ratio 3.5:1) was noted; among the six women who have been pregnant at least once, three experienced important clinical changes during pregnancy. No germ-line mutation was found in the exons 10, 11, and 17 of the KIT proto-oncogene, which are the most commonly mutated exons in sporadic mastocytosis. However, in the majority of affected subjects we found the Met541Leu polymorphic variant of the KIT gene, which seems to confer a growth advantage to mast cells in vitro. This observation further suggests that the Met541Leu may be a predisposing factor of cutaneous mastocytosis, although it seems to be neither necessary nor sufficient for the development of the disease.
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Tourlaki, Athanasia <1973>. "The KIT gene in familial mastocytosis." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2013. http://amsdottorato.unibo.it/5590/.
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Familial cutaneous mastocytosis is an exceptional condition of unknown etiology. In this study we report the largest series of patients with familial cutaneous mastocytosis without other manifestations (18 affected subjects from seven unrelated families), and we investigate the role of germ-line KIT mutations in the pathogenesis of the disease. The mean age at onset was 5.4 years (range from birth to 22 years), and the clinical behavior was variable over a mean follow up period of 15.1 years (range 2-36): improvement in seven, stability in eight and worsening in the remaining three patients. The pattern of inheritance was compatible with an autosomal dominant trait with incomplete penetrance; a female preponderance (14 females vs 4 males, ratio 3.5:1) was noted; among the six women who have been pregnant at least once, three experienced important clinical changes during pregnancy. No germ-line mutation was found in the exons 10, 11, and 17 of the KIT proto-oncogene, which are the most commonly mutated exons in sporadic mastocytosis. However, in the majority of affected subjects we found the Met541Leu polymorphic variant of the KIT gene, which seems to confer a growth advantage to mast cells in vitro. This observation further suggests that the Met541Leu may be a predisposing factor of cutaneous mastocytosis, although it seems to be neither necessary nor sufficient for the development of the disease.
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Aramini, Beatrice <1979>. "Role of SP-A gene polymorphism in lung transplantation." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2011. http://amsdottorato.unibo.it/3634/1/aramini_beatrice_tesi.pdf.
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Lung transplantation is a widely accepted therapeutic option for end stage lung disease. Clinical outcome is yet challenged by primary graft failure responsible for the majority of the early mortality, by chronic allograft dysfunction and chronic rejection accounting for more than 30% of deaths after the third postoperative year. Pulmonary surfactant proteins (SP) A, B, C and D are one of the first host defense mechanisms the lung can mount. SP-A in particular, produced by the type II pneumocytes, is active in the innate and adaptive immune system being an opsonin, but also regulating the macrophage and lymphocyte response. The main hypothesis for this project is that pulmonary surfactant protein A polymorphism may determine the early and long term lung allograft survival. Of note SP-A biologic activity seems to be genetically determined and SP-A polymorphisms have been associated to various lung disease. The two SP-A genes SP-A1 and SP-A2 have several polymorphisms within the coding region, SP-A1 (6A, 6A2-20), and SP-A2(1A, 1A0-13). The SP-A gene expression is regulated by cAMP, TTF-1 and glucocorticoids. In vitro studies have indicated that SP-A1 and SP-A2 gene variants may have a variable response to glucocorticoids. We proposed to determine if SP-A gene polymorphism predicts primary graft dysfunction and/or chronic lung allograft dysfunction and if SP-A may serve as a biomarker of lung allograft dysfunction. We also proposed to study the interaction between immunosuppressive drugs and SP-A expression and determine whether this is dependent on SP-A polymorphisms. This study will generate novel information improving our understanding of lung allograft dysfunction. It is conceivable that the information will stimulate the interest for a multi centre study to investigate if SP-A polymorphism may be integrated in the donor lung selection criteria and/or to implement post transplant tailored immunosuppression.
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Aramini, Beatrice <1979>. "Role of SP-A gene polymorphism in lung transplantation." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2011. http://amsdottorato.unibo.it/3634/.
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Lung transplantation is a widely accepted therapeutic option for end stage lung disease. Clinical outcome is yet challenged by primary graft failure responsible for the majority of the early mortality, by chronic allograft dysfunction and chronic rejection accounting for more than 30% of deaths after the third postoperative year. Pulmonary surfactant proteins (SP) A, B, C and D are one of the first host defense mechanisms the lung can mount. SP-A in particular, produced by the type II pneumocytes, is active in the innate and adaptive immune system being an opsonin, but also regulating the macrophage and lymphocyte response. The main hypothesis for this project is that pulmonary surfactant protein A polymorphism may determine the early and long term lung allograft survival. Of note SP-A biologic activity seems to be genetically determined and SP-A polymorphisms have been associated to various lung disease. The two SP-A genes SP-A1 and SP-A2 have several polymorphisms within the coding region, SP-A1 (6A, 6A2-20), and SP-A2(1A, 1A0-13). The SP-A gene expression is regulated by cAMP, TTF-1 and glucocorticoids. In vitro studies have indicated that SP-A1 and SP-A2 gene variants may have a variable response to glucocorticoids. We proposed to determine if SP-A gene polymorphism predicts primary graft dysfunction and/or chronic lung allograft dysfunction and if SP-A may serve as a biomarker of lung allograft dysfunction. We also proposed to study the interaction between immunosuppressive drugs and SP-A expression and determine whether this is dependent on SP-A polymorphisms. This study will generate novel information improving our understanding of lung allograft dysfunction. It is conceivable that the information will stimulate the interest for a multi centre study to investigate if SP-A polymorphism may be integrated in the donor lung selection criteria and/or to implement post transplant tailored immunosuppression.
10
SATTA, STEFANIA. "Studio del gene AHSP nelle β-Talassemie." Doctoral thesis, Università degli Studi di Cagliari, 2007. http://hdl.handle.net/11584/265981.
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Objective: To establish whether AHSP might have a role in modifying the clinical severity of beta-thalassemia.
Background: The severity of beta-thalassemia reflects the degree of globin chain imbalance and the excess of free alpha-globin chains that precipitate and cause oxidative damage in red cell precursors, inducing their premature destruction (ineffective erythropoiesis). AHSP is an abundant erythroid-specific protein that binds specifically to free alpha-globin and prevents its precipitation in vitro. It has been demonstrated in the animal model that AHSP is required for normal haemoglobin production and that it acts as a chaperone, to prevent the harmful aggregation of alpha-globin during normal erythroid development and in diseases with globin chain imbalance.
Patients and methods: AHSP gene (~1.4 Kb) and its promoter region was directly sequenced from PCR amplified DNA on ABI PRISM 3100, in 19 thalassemia intermedia and 14 thalassemia major patients with similar genetic characteristics (beta39/beta39, 3 or 4 alpha-globin genes and I-II or II-II haplotype according to Orkin et al).
Results: No nucleotide mutation that might alter the structure and function of AHSP was identified. We found in the large majority of the cases the same haplotypes with the same frequencies described in S/E Asia (Viprakasit et al., Blood 2004). No difference has been found in the haplotype frequencies between the beta-thalassemia major and intermedia patients.
Conclusion: We did not find mutations or specific association between haplotype of AHSP and disease severity in these patients, suggesting that AHSP is not a disease modifier in Sardinian beta-thalassemia patients.
11
Zattra, Edoardo. "Analisi di polimorfismi genetici del gene TP53 e del gene EGF in pazienti con nevi melanocitici multipli." Doctoral thesis, Università degli studi di Padova, 2011. http://hdl.handle.net/11577/3422737.
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Background: p53 have been extensively reported in the literature to be able to modify the activity of melanocytes, particularly in controlling the proliferation of these cells. P53 is a transcriptional activator of genes encoding for proteins that influence the proliferation of melanocytes and in this way the onset and progression of malignant melanoma. Epidermal Growth Factor (EGF) is a growth factor member of the EGF superfamily. It has been shown that it activates cell proliferation and stimulates mitogenesis in epidermal tissue, enhancing tumor growth. The presence of more than 100 nevi has been demonstrated to be a major risk factor for developing malignant melanoma.
Aim of the study: Several experimental data indicate that the two genetic polymorphism of EGF and P53 (IVS6 +62 G> A in intron 6 of TP53 and EGF +61A>G) could play a role in the onset and progression of several neoplasms. We studied the above-mentioned gene polymorphisms in a population of patients with more than 100 melanocytic nevi. Patients were carefully selected on the basis of an accurate clinical and dermatoscopic examination.
Materials and methods: As regards the IVS6 +62 G> A in intron 6 of TP53 polymorphism 98 patients and 117 controls were investigated while for the EGF +61A>G polymorphism we included in the study 128 patients and 127 controls. Patients were all aged between 21 and 60 years and had an high number of melanocytic nevi (>100). Controls were patients with a low number of nevi (< 10) of the same sex and age. For the analysis of the polymorphisms DNA was extracted from peripheral blood and genotyped by High Resolution Melt Analysis (HRM).
Results:
For p53, the genotype (A/A) was present in 4.2% of the patients against a rate of 1.9% in controls. Genotype G/A was significantly increased in patients (32.5%) compared to controls (23.6%). With regard to allele frequencies, G allele was present in 79.6% of patients and 86.3% of controls whereas A allele was present in 20,4% of patients and 13.7% of controls. For EGF, genotype AA was present in 35% of patients and 38% of controls. Genotype A/G was present in 48% of patients and 44% of controls while genotype G/G was present in 17% of patients and 18% of controls. With regard to allele frequency, A was present in 59% of patients and in 60% of controls, whereas G was observed in 41% of patients and 40% of controls.
Conclusions: The polymorphism IVS6 +62 G> A of the TP53 gene was associated with the presence of more than 100 melanocytic nevi indicating that alterations in p53 expression, even if minimal as in this polymorphism, could cause an altered control of melanocytic proliferation that clinically reflects in an high number of melanocytic nevi. There was no significative correlation between the EGF +61A>G polymorphism and the presence of a high number of melanocytic nevi.Background: p53 è in grado di modificare l’attività dei melanociti, in particolare la proliferazione di queste cellule. P53 è un attivatore della trascrizione di geni che codificano per proteine con azione stimolatoria sulla proliferazione dei melanociti, pertanto in questo modo p53 può favorire l’insorgenza e la progressione del melanoma maligno. L'Epidermal Growth Factor (EGF) è un fattore di crescita membro della superfamiglia EGF. E’ stato dimostrato che l'EGF attiva la proliferazione cellulare e stimola la mitogenesi nel tessuto cutaneo, stimolando la crescita tumorale. E’ stato dimostrato che la presenza di più di 100 nevi melanocitici è un fattore di rischio importante per la comparsa del melanoma maligno. Scopo dello studio: I polimorfismi genetici presi in esami in questo studio (IVS6 +62 G> A all’introne 6 di TP53 ed EGF +61A>G e) secondo numerosi studi potrebbero giocare un ruolo nell’insorgenza e nella progressione di diverse neoplasie. Abbiamo analizzato i suddetti polimorfismi in una popolazione di pazienti con più di 100 nevi melanocitici mettendo tali dati a confronto con una popolazione di controllo con meno di 10 nevi melanocitici. Tali pazienti sono stati accuratamente selezionati attraverso esami clinici e dermatoscopici. Materiali e Metodi: Per lo studio di p53 abbiamo incluso 98 pazienti e 117 controlli, mentre per lo studio dell' EGF abbiamo incluso 128 pazienti e 127 controlli tutti di età compresa tra i 21 e i 60 anni. I pazienti presentavano più di 100 nevi melanocitici di dimensioni superiori a 3 millimetri di diametro. I controlli erano soggetti con meno di 10 nevi melanocitici. Per l’analisi del polimorfismo il DNA è stato estratto da sangue periferico e il genotipo è stato studiato mediante High Resolution Melt Analysis (HRM). Risultati: Il genotipo A/A di p53, era presente nel 4.2% dei pazienti contro il 1.9% dei controlli. Il genotipo G/A era significativamente maggiore nei pazienti (32.5%) che nei controlli (23.6%). Per quanto riguarda le frequenze alleliche, l’allele G era presente nel 79.6% dei pazienti e nel 86.3% dei controlli mentre l’allele A era presente nel 20,4% dei pazienti e nel 13,7% dei controlli. Per quanto riguarda EGF, il genotipo A/A era presente nel 35% dei pazienti e nel 38% dei controlli. Il genotipo A/G era presente nel 48% dei pazienti e nel 44% dei controlli mentre il genotipo G/G era presente nel 18% dei pazienti e nel 17% dei controlli. Per quanto riguarda la frequenza allelica, A era presente nel 59% dei pazienti e nel 60% dei controlli, mentre G era presente nel 41% dei pazienti e nel 40% dei controlli. Conclusioni: non è stata rilevata una correlazione significativa tra il polimorfismo EGF +61A>G e la presenza di un alto numero di nevi melanocitici. Il polimorfismo IVS6 +62 G> A del gene TP53 sembra invece essere associato con la presenza di un elevato numero di nevi melanociti essendo la frequenza di questo polimorfismo significativamente maggiore nei casi rispetto ai controlli. E’ plausibile che alterazioni dell’espressione della p53, sia pur minime come quelle osservabili nei polimorfismi genetici che coinvolgono regioni regolatorie del DNA, possano essere responsabili di un alterato controllo della proliferazione melanocitaria che si traduce clinicamente in un alto numero di nevi melanocitici.
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Caraffi, Stefano Giuseppe <1977>. "Analysis of TNFRSF5 gene mutations and splicing variants in CD40 receptor regulation." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2007. http://amsdottorato.unibo.it/605/1/Caraffi_tesi.pdf.
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Caraffi, Stefano Giuseppe <1977>. "Analysis of TNFRSF5 gene mutations and splicing variants in CD40 receptor regulation." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2007. http://amsdottorato.unibo.it/605/.
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Rafferty, John Bernard. "X-ray crystallographic structure determination of the Met repressor from Escherichia coli and its functional implications." Thesis, University of Leeds, 1990. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.255349.
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Andic, Berna. "Lärares arbetssätt med olika texttyper i svenskämnet, med inriktning mot årskurs 4-6." Thesis, Södertörns högskola, Lärarutbildningen, 2019. http://urn.kb.se/resolve?urn=urn:nbn:se:sh:diva-39844.
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The purpose of this study is to investigate how four teachers in grades 4-6 work with genre pedagogy in the Swedish subject. In this study the focus will be on these two issues: How do the teachers work with text types in the Swedish subject? How do the teachers describe genre pedagogy? I have used the two different methods which are interviews and observations in order to conduct the survey. I have also used two different theories that are related to the genre pedagogy in this study, these theories are the socio-cultural perspective and the circle model. These theories have been used to analyze the empirical material in this study. The result of this study shows that all four teachers work with the four different types of texts which are fact text, story, fabel and instruction in teaching in the Swedish subject. Furthermore all the teachers work in the same way with these different text types in the teaching, this by working with the text types based on the four different phases that are in the circle model in the teaching. These different steps consist of knowledge acquisition of a subject, the study of the text type with focus on the purpose, structure and linguistic features of the text type, the class writing its own text within the chosen text type, which is followed by the students being allowed to write their own text within the chosen text type. This was reflected in the interviews and two of the observations. All teachers shared the same view that genre pedagogy is about a number of different genres. The texts that are written within a genre should consist of a specific structure, and language, depending on the purpose, situation and recipients of the texts, according to the teachers. This was evident in the interviews in this study.
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Gennari, Monia <1972>. "Analisi del gene PRKA1A in una famiglia affetta da Carney Complex." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2007. http://amsdottorato.unibo.it/149/1/tesiDottoratoGennari.pdf.
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Gennari, Monia <1972>. "Analisi del gene PRKA1A in una famiglia affetta da Carney Complex." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2007. http://amsdottorato.unibo.it/149/.
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CARRIGLIO, NICOLA. "Preclinical gene therapy studies, altered lymphocyte development and function in ADA-SCID." Doctoral thesis, Università degli Studi di Roma "Tor Vergata", 2012. http://hdl.handle.net/2108/209654.
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Genetic defects in the adenosine deaminase (ADA) gene are among the most
common causes for severe combined immunodeficiency (SCID). ADA-SCID
patients suffer from lymphopenia, absent cellular and humoral immunity,
recurrent infections and autoimmune manifestations in milder forms. Currently
available therapeutic options for this otherwise fatal disorder include bone
marrow transplantation (BMT), enzyme replacement therapy with bovine ADA
(PEG-ADA) or hematopoietic stem cell gene therapy (GT). The overall aims of
my this thesis were to evaluate the preclinical safety of HSC gene therapy and
the basis involved in autoimmune manifestations observed in ADA deficiency.
For the first part of the project I assessed the feasibility to perform two preclinical
studies recommended by Regulatory Authorities:
toxicology/tumorigenicity and biodistribution studies. Both studies were
performed using the NSG immunodeficient mouse model transplanted with
murine or human hematopoietic stem/progenitor cells transduced with a
Retroviral vector with an amphotrophic envelope encoding for ADA. For the
toxicology/tumorigenicity study we tested different transduction protocols both in
vitro and in vivo showing a low transduction efficiency in ADA-/- mouse lineage
negative cells. These results led us to conclude that this model is not suitable
for in vivo toxicology/tumorigenicity studies. On the other hand, transduction
efficiency in human hematopoietic stem/progenitor cells was higher compared
to murine cells. Biodistribution study with human CD34+ cells derived from cord
blood showed a good engraftment of human cells in NSG hosts with transduced
cells in PB and lymphoid organs, in line with the frequency found in treated patients.The second part of my PhD project aimed at investigating Treg function and B
cell development in ADA-deficient mice. Autoimmune manifestations including
type I diabetes, hypothyroidism, autoimmune thrombocytopenia, and haemolytic
anaemia are frequently observed in the ADA-SCID patients treated with PEGADA,
BMT and GT. We investigated the mechanisms which might be involved
in these alterations. We found that PEG_ADA treated mice represent a model to
study autoimmunity as they developed multiple autoantibodies and
hypothyroidism in contrast to mice treated with bone marrow transplantation or
gene therapy. Moreover, Tregs isolated from PEG-ADA– treated mice lacked
suppressive activity, suggesting that this treatment interferes with Treg
functionality.
mice showed a mild alteration in the bone marrow and splenic B cell subsets.
We also explored whether an increased egress of immature and recirculant B
cells from the bone marrow through the synovial vessels due to the activation of
endocannabinoid pathway might contribute to autoimmune manifestations in
this disease. B-cell escaping central tolerance mechanisms together with nonfunctional
Treg cells in the periphery might further accelerate the onset of
autoimmunity.
In summary results of this work have contributed to improved our knowledge on
ADA-SCID and facilitate the progress of clinical development of gene therapy for ADA-SCID.
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BOTTANI, EMANUELA. "Mitochondrial diseases: from gene function to therapy." Doctoral thesis, Università degli Studi di Milano-Bicocca, 2015. http://hdl.handle.net/10281/94380.
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Le malattie mitocondriali sono disturbi genetici caratterizzati da difetti di fosforilazione ossidativa causati da mutazioni nel DNA mitocondriale, o in geni nucleari i cui prodotti sono legati alla fosforilazione ossidativa o alla biologia mitocondriale. La prima parte del progetto è stata focalizzata sulla generazione e la caratterizzazione di un modello murino di malattia mitocondriale, Ttc19ko. I pazienti con mutazioni in TTC19 sviluppano danni neurologici e deficit di complesso III. Ttc19 è una proteina mitocondriale che sembra essere coinvolta nell’assemblaggio e/o stabilità del complesso III. Abbiamo dimostrato che il modello murino Ttc19ko ha sintomi neurologici, debolezza muscolare e riduzione dell’attività locomotoria spontanea, in analogia con la malattia umana. L’analisi istologica ha rivelato alcune anomalie neurologiche con presenza di accumuli di ubiquitina e GFAP. I topi Ttc19ko hanno una riduzione complessiva del metabolismo energetico, una diminuzione del consumo di O2 e di produzione di CO2. L’attività enzimatica del complesso III è significativamente ridotta nei tessuti e ciò è legato ad un aumento della produzione di ROS. L’analisi BNGE ha mostrato una riduzione della incorporazione della subunità catalitica Rieske nel complesso assemblato. L’immunoprecipitazione di TTC19-Flag in colture cellulari trattate con amminoacidi marcati ha rivelato una maggiore interazione tra Ttc19 e le subunità del pre-complesso III, e una minore interazione con proteine Rieske e Uqcrh, entrambe assemblate tardivamente. Abbiamo inoltre dimostrato che Ttc19 rimane associata al complesso III assemblato. Nell’insieme, questi risultati suggeriscono che Ttc19 è un fattore intrinseco di assemblaggio del complesso III che interagisce con il pre-complesso III facilitando così l'incorporazione della proteina Rieske. La seconda parte del progetto è stata focalizzata sulla messa a punto di una terapia genica su un altro modello murino di malattia mitocondriale, MPV17ko. Mutazioni in MPV17 causano una sindrome epatocerebrale con deplezione del mtDNA, insufficienza epatica a esordio precoce, gravi crisi ipoglicemiche e morte. Il trapianto di fegato e l'alimentazione frequente a base di carboidrati a lento rilascio sono le uniche terapie disponibili, anche se in seguito si sviluppano danni neurologici. Il ruolo fisiologico di MPV17 non è chiaro. Abbiamo dimostrato che MPV17 fa parte di un complesso ad alto peso molecolare a composizione sconosciuta, che è essenziale per il mantenimento del mtDNA nel fegato. In dieta standard, il topo MPV17-/- non mostra quasi alcun sintomo di disfunzione epatica, ma una dieta chetogenica porta questi animali a sviluppare cirrosi e insufficienza epatica grave. Tuttavia, quando l'espressione di MPV17 è ripristinata dalla somministrazione di virus adeno-associato, si assiste ad un ricostituzione del complesso supramolecolare contenente Mpv17, ad un ripristino completo del numero di copie di mtDNA, ed alla prevenzione dell’insufficienza epatica indotta dal dieta chetogenica. Questi risultati aprono nuove prospettive terapeutiche per il trattamento delle sindromi da deplezione del mtDNA indotte da mutazioni nel gene MPV17.Mitochondrial diseases are genetic disorders characterized by defects in oxidative phosphorylation caused by mutations in mitochondrial DNA, or in nuclear genes whose products are related to oxidative phosphorylation or mitochondrial biology. The first part of the project was focused on the generation and characterization of a mouse model of mitochondrial disease, Ttc19ko. Patients with mutations in TTC19 were characterized by neurological impairments and mitochondrial respiratory complex III deficiency. Ttc19 is a mitochondrial protein that seems to be associated to complex III assembly and/or stability. We showed that Ttc19ko mice have neurological symptoms, muscular weakness and reduction in spontaneous locomotors activity, clearly resembling the human disease. Brain also had neurological abnormalities with presence of ubiquitin and GFAP positive staining. Comprehensive lab animals monitoring system revealed a reduction in O2 consumption, CO2 production and energy expenditure in Ttc19ko mice, indicating an overall reduction of energy metabolism. Complex III activity was significantly reduced in tissues and this was linked to an increased ROS production. BNGE analysis of mitochondrial complex III showed a substantial reduction of the incorporation of the catalytic Rieske iron-sulfur protein into the fully assembled complex. A stable isotope labelling by amino acids in cell culture (SILAC) expressing TTC19-Flag followed by immunoprecipitation and mass spec analysis revealed a higher scored interaction between Ttc19 and the subunits of the pre-complexIII, and a lower scored interaction with Rieske protein and Uqcrh, both of them are late assembled subunits. We also demonstrated that Ttc19 is associated to the fully assembled complex III. Taken together, these results suggests that Ttc19 is an intrinsic assembly factor of complex III that interacts with the pre-complex III thus facilitating the incorporation of the late assembled Rieske protein. The second part of the project was focused on a gene therapy approach on a second mouse model of mitochondrial disease, MPv17ko. Mutations in hMPV17 cause a hepatocerebral form of mtDNA depletion syndrome hallmarked by early-onset liver failure, leading to premature death. Liver transplantation and frequent feeding using slow-release carbohydrates are the only available therapies, although surviving patients develop slowly progressive neuropathy. The physiological role of Mpv17 is still unclear. We showed that Mpv17 is part of a high molecular weight complex of unknown composition, which is essential for mtDNA maintenance in liver. On a standard diet, Mpv17ko mouse shows hardly any symptom of liver dysfunction, but a ketogenic diet leads these animals to liver cirrhosis and failure. However, when expression of human MPV17 is carried out by adeno-associated virus mediated gene replacement, the Mpv17ko mice are able to reconstitute the Mpv17-containing supramolecular complex, restore liver mtDNA copy number and oxidative phosphorylation proficiency and prevent liver failure induced by the KD. These results open new therapeutic perspectives for the treatment of MPV17-related liver-specific MDS.
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LATINA, ALESSIA. "Cytoglobin, a direct ΔNp63 target gene, counteracts oxidative stress in keratinocytes." Doctoral thesis, Università degli Studi di Roma "Tor Vergata", 2012. http://hdl.handle.net/2108/209986.
Full textAbstract:
p63 è un fattore di trascrizione appartenente alla famiglia di p53 insieme
a p53 e p73. Essi presentano un certo grado di omologia ma non sono
funzionalmente ridondanti. P63, and in particolare l’isoforma ΔNp63α
svolge un ruolo importante nello sviluppo epidermico (proliferazione e
differenziazione). Il suo livello di espressione varia nei diversi strati
dell'epidermide, è abbondante nello strato basale e diminuisce negli strati
superiori differenziati. L’epidermide è un tessuto fisiologicamente soggetto a
stress ossidativo. La produzione di specie reattive dell'ossigeno (ROS) può
originare da varie fonti, possono essere prodotti nel corso del normale
metabolismo aerobico della cellula oppure derivare dall’ambiente
circostante. Studi recenti hanno rivelato un coinvolgimento di p63 nella
risposta allo stress ossidativo. Allo scopo di identificare nuovi target di p63
nella risposta antiossidante, dopo aver osservato un aumento dei ROS
intracellulari a seguito dell' inibizione di p63, abbiamo utilizzato un PCR
array per analizzare l'espressione di una serie di geni appartenenti al
metabolismo dei ROS. Questo metodo ci ha consentito di analizzare
contemporaneamente 84 geni riguardanti lo stress ossidativo e la difesa
antiossidante. In particolare, dopo il silenziamento di TP63, 11 geni
presentavano una espressione ridotta e 5 aumentata. Ci siamo focalizzati sul
gene CYGB, che codifica per la citoglobina, un membro della famiglia delle
globine che facilitano la diffusione di ossigeno attraverso i tessuti e agiscono
come scavenger di ossido nitrico o altre specie reattive dell'ossigeno.
Attraverso saggi di attività del promotore, immunoprecipitazione della
cromatina (ChIP), real-time PCR e analisi western blot abbiamo confermato
la regolazione diretta della CYGB da parte di ΔNp63α. Esperimenti di
silenziamento della citoglobina hanno mostrato che i cheratinociti risultano
più sensibili allo stress ossidativo e che se trattati con perossido di idrogeno
mostrano un aumento dei livelli di ROS e di apoptosi rispetto al controllo. I
dati quindi indicano che p63, attraverso il suo target trascrizionale CYGB, ha
un ruolo importante nel proteggere i cheratinociti dallo stress ossidativo.p63 is a transcription factor belonging to p53 family. The members of this family share a substantial degree of homology but they are not functionally redundant. The ΔNp63α isoform plays a critical role in the formation of the epidermis, its expression level varies in the different layers of the epidermis, it is abundant in the basal proliferating layer and decreases in the upper differentiated layers. The epidermis undergoes to oxidative stress by production of reactive oxygen species (ROS) in a physiological manner, during normal aerobic metabolism. ROS also derive from the environment, ie. UV irradiation. In order to maintain the balance of the oxidative status, cells have protective antioxidant systems. Preliminary studies suggest that p63 could be involved in the oxidative defense. To identify p63 targets, possibly involved in the anti-oxidant defence, we have used t RT2 Profiler PCR Array. After TP63 silencing, we found 16 genes modulated, and we focused our attention on one gene, CYGB. CYGB codes for cytoglobin, a member of the globin protein family that facilitates diffusion of oxygen through tissues and acts as a scavenger for nitric oxide or other reactive oxygen species. We performed promoter activity assay, chromatin immunoprecipitation (ChIP) assay, real-time PCR and western blot analysis to confirm the direct regulation of CYGB by ΔNp63α. We also performed studies to evaluate the biological role of cytoglobin in keratinocytes. In particular, we detected an increase of cell death and ROS levels in keratinocytes depleted of CYGB, this increase was even more consistent upon hydrogen peroxide treatments. The results obtained indicate that p63, and its target gene CYGB, has an important role in protecting keratinocytes from oxidative stress.
21
FAVASULI, VANESSA KATIA. "DISSECTING THE CLINICAL AND BIOLOGICAL RELEVANCE OF DIS3 GENE IN MULTIPLE MYELOMA." Doctoral thesis, Università degli Studi di Milano, 2023. https://hdl.handle.net/2434/956171.
Full textAbstract:
Multiple myeloma (MM) is a B cell malignancy characterized by abnormal proliferation of plasma cells (PCs) within the bone marrow. MM is characterized by a wide clinical spectrum ranging from the presumed asymptomatic pre-malignant condition called monoclonal gammopathy of undetermined significance (MGUS), to extra-medullary plasma cell leukemia (PCL). Notably, MM is characterized by a deep genomic instability involving ploidy, structural rearrangements and a high range of mutations involving both putative oncogenes and tumor suppressor genes that may explain its clinical heterogeneity. Half of MM tumors are hyperdiploid, associated with non-random trisomies of odd chromosomes and low prevalence of chromosomal translocations involving the immunoglobulin heavy chain locus (IGH) on chromosome 14q32, whereas the remaining tumors are non hyperdiploid and characterized by the prevalence of IGH translocations affecting several oncogenic loci. Such a large heterogeneous genomic pattern may represent the basis for the abnormal transcriptional expression profile associated with MM.
Despite the remarkable improvements in treatment and patient care, MM remains an incurable disease.
Over the past years, our and other research groups have provided valuable information on coding and non-coding transcripts aberrantly expressed in MM by expression profiling analyses based on microarray or RNAseq approaches. These studies have been of high relevance to better understand the biology of the disease, to identify of novel prognostic and predictive biomarkers and putative therapeutic targets.
DIS3 (chr.13q22.1 localization) is a conserved exoribonuclease and catalytic subunit of the exosome, a protein complex involved in the 3' to 5' degradation and processing of different species of RNA. Recently, aberrant expression of DIS3 has been found to be implicated in a range of different cancers. Notably, DIS3 is recurrently mutated in multiple myeloma (MM) patients. Most of the identified mutations are predominantly missense variants localized in the ribonucleolytic domain (RNB), mainly abolishing the exoribonucleolytic activity and are often accompanied by loss of heterozygosity (LOH) or biallelic inactivation due to 13q14 deletion. Furthermore, it has been reported in the literature that the inactivity or incorrect activity of DIS3 is also associated with the regulation of non-coding transcripts, as well as long non-coding RNA (lncRNA).
Long non-coding RNAs (lncRNAs) are a large class of non-coding RNAs involved in many physiological cellular and genomic processes as well as in carcinogenesis, cancer metastasis and invasion. The knowledge of the role of lncRNAs in MM is progressively expanding. Our group provided recent evidence based on microarray and RNA seq analyses of deregulated patterns of lncRNA expression in MM showing that they may be specifically associated with distinct molecular types of the malignancy.
In a paper published during the first years of the project, it has been shown that MM patients carrying Dis3 gene mutations are associated with a distinct transcriptional signature characterized by many non-coding transcripts, mainly lncRNAs.
Firstly, our interest focused on investigating the deregulation of transcripts (including coding and non-coding portion) associated with DIS3 mutations in MM by analyzing RNA-Seq transcriptional profiles in a proprietary publicly available dataset and CoMMpass study, including approximately 1000 MM patients at diagnosis. We demonstrated that DIS3 mutations clinical relevance strictly depended on del(13q) co-occurrence. We observed that the bi-allelic DIS3 lesions significantly affected PFS (Progression Free Survival) and overall survival (OS). As expected, DIS3 mutations affect MM transcriptome involving cellular processes and signaling pathways associated with RNA metabolism. We found the downregulation of gene sets related to oxidative phosphorylation, RNA or amino acid metabolism, translation, and mitotic spindle checkpoint.
Overall, our comprehensive assessment of the clinical and transcriptional consequences of DIS3 mutations or its deletion in MM strongly indicates that they may play an important role in the mechanisms of transformation and progression of MM.
For these reasons, during the last year of my PhD program I focalized the efforts on the functional investigation of DIS3 putative role as potential therapeutic target in MM. To this aim I took advantage of the experimental silencing strategy based on the use of LNA-gapmeR technology and the gymnotic delivery of the in-house designed DIS3-specific antisense oligonucleotide in multiple myeloma cell lines (HMCLs).
The results obtained show that DIS3 silencing decreases cell growth, increases the percentage of apoptosis, reducing the oncogenic potential of MM cell lines. Indeed, interference with DIS3 expression reveals an important perturbation of the cell cycle distribution accompanied by mitotic defects.
I investigated the possibility of translating these results in combination with an available drug currently in clinical trial, such as ARRY-520, an inhibitor that interferes with the correct organization of microtubules. I found that the effect of DIS3 KD sensitizes MM cell lines in combination with ARRY-520, leading to a mitotic catastrophe.
These results suggest that DIS3 could be an important target for the future therapeutic approach in MM disease.
22
Del, Sorbo Francesca <1970>. "Caratterizzazione dei sintomi neurovegetativi e neuropsicologici nella malattia di Parkinson associata a mutazioni del gene glucocerebrosidasi." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2015. http://amsdottorato.unibo.it/6956/1/Del_Sorbo_Francesca_Eddi_Alice_tesi.pdf.
Full textAbstract:
Pochi studi hanno indagato il profilo dei sintomi non-motori nella malattia di Parkinson associata al gene glucocerebrosidasi (GBA). Questo studio è mirato alla caratterizzazione dei sintomi non-motori, con particolare attenzione alla valutazione delle funzioni neurovegetativa, cognitiva e comportamentale, nel parkinsonismo associato a mutazione del gene GBA con la finalità di verificare se tali sintomi non-motori siano parte dello spettro clinico di questi pazienti.
E’ stato condotto su una coorte di pazienti affetti da malattia di Parkinson che erano stati tutti sottoposti ad una analisi genetica per la ricerca di mutazioni in uno dei geni finora associati alla malattia di Parkinson. All’interno di questa coorte omogenea sono stati identificati due gruppi diversi in relazione al genotipo (pazienti portatori della mutazione GBA e pazienti non portatori di nessuna mutazione) e le caratteristiche non-motorie sono state confrontate nei due gruppi. Sono state pertanto indagati il sistema nervoso autonomo, mediante studio dei riflessi cardiovascolari e analisi dei sintomi disautonomici, e le funzioni cognitivo-comportamentali in pazienti affetti da malattia di Parkinson associata a mutazione del gene GBA. I risultati sono stati messi a confronto con il gruppo di controllo.
Lo studio ha mostrato che i pazienti affetti da malattia di Parkinson associata a mutazione del gene GBA presentavano maggiore frequenza di disfunzioni ortosimpatiche, depressione, ansia, apatia, impulsività, oltre che di disturbi del controllo degli impulsi rispetto ai pazienti non portatori.
In conclusione, i pazienti GBA positivi possono esprimere una sintomatologia non-motoria multidominio con sintomi autonomici, cognitivi e comportamentali in primo piano. Pertanto l’impostazione terapeutica in questi pazienti dovrebbe includere una accurata valutazione dei sintomi non-motori e un loro monitoraggio nel follow up clinico, allo scopo di ottimizzare i risultati e ridurre i rischi di complicazioni.Few studies have investigated the non-motor symptoms profile in Parkinson disease (PD) associated with the glucocerebrosidase gene (GBA). This study is aimed at characterizing non-motor features, with particular attention to the evaluation of autonomic, cognitive and behavioral functions, in PD associated with mutations in the GBA gene with the aim to verify if these symptoms are part of the clinical spectrum of these patients. A study has been conducted on a cohort of patients with PD who had all been subjected to genetic analysis for the detection of mutations in one of the genes so far associated with PD. Within this homogeneous cohort, two different groups were identified in relation to the genotype (patients carriers of GBA mutation and patients noncarriers of genes associated with PD) and the non-motor characteristics were compared in the two groups. We have therefore investigated the autonomic nervous system, through the study of cardiovascular reflexes and analysis of autonomic symptoms, and cognitive-behavioral functions in patients with PD associated with mutations in the GBA gene. The results were compared with the control group. The study showed that patients with PD associated with mutations in the GBA gene had higher frequency of sympathetic dysfunction, depression, anxiety, apathy, impulsivity, as well as disorders of impulse control compared to noncarriers patients. In conclusion, patients GBA positive can manifest a multidomain non-motor symptom profile with autonomic, cognitive and behavioral symptoms. Therefore, the therapeutic approach in these patients should include a thorough assessment of non-motor symptoms and their monitoring in the follow-up, in order to optimize the results and reduce the risk of complications.
23
Del, Sorbo Francesca <1970>. "Caratterizzazione dei sintomi neurovegetativi e neuropsicologici nella malattia di Parkinson associata a mutazioni del gene glucocerebrosidasi." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2015. http://amsdottorato.unibo.it/6956/.
Full textAbstract:
Pochi studi hanno indagato il profilo dei sintomi non-motori nella malattia di Parkinson associata al gene glucocerebrosidasi (GBA). Questo studio è mirato alla caratterizzazione dei sintomi non-motori, con particolare attenzione alla valutazione delle funzioni neurovegetativa, cognitiva e comportamentale, nel parkinsonismo associato a mutazione del gene GBA con la finalità di verificare se tali sintomi non-motori siano parte dello spettro clinico di questi pazienti.
E’ stato condotto su una coorte di pazienti affetti da malattia di Parkinson che erano stati tutti sottoposti ad una analisi genetica per la ricerca di mutazioni in uno dei geni finora associati alla malattia di Parkinson. All’interno di questa coorte omogenea sono stati identificati due gruppi diversi in relazione al genotipo (pazienti portatori della mutazione GBA e pazienti non portatori di nessuna mutazione) e le caratteristiche non-motorie sono state confrontate nei due gruppi. Sono state pertanto indagati il sistema nervoso autonomo, mediante studio dei riflessi cardiovascolari e analisi dei sintomi disautonomici, e le funzioni cognitivo-comportamentali in pazienti affetti da malattia di Parkinson associata a mutazione del gene GBA. I risultati sono stati messi a confronto con il gruppo di controllo.
Lo studio ha mostrato che i pazienti affetti da malattia di Parkinson associata a mutazione del gene GBA presentavano maggiore frequenza di disfunzioni ortosimpatiche, depressione, ansia, apatia, impulsività, oltre che di disturbi del controllo degli impulsi rispetto ai pazienti non portatori.
In conclusione, i pazienti GBA positivi possono esprimere una sintomatologia non-motoria multidominio con sintomi autonomici, cognitivi e comportamentali in primo piano. Pertanto l’impostazione terapeutica in questi pazienti dovrebbe includere una accurata valutazione dei sintomi non-motori e un loro monitoraggio nel follow up clinico, allo scopo di ottimizzare i risultati e ridurre i rischi di complicazioni.Few studies have investigated the non-motor symptoms profile in Parkinson disease (PD) associated with the glucocerebrosidase gene (GBA). This study is aimed at characterizing non-motor features, with particular attention to the evaluation of autonomic, cognitive and behavioral functions, in PD associated with mutations in the GBA gene with the aim to verify if these symptoms are part of the clinical spectrum of these patients. A study has been conducted on a cohort of patients with PD who had all been subjected to genetic analysis for the detection of mutations in one of the genes so far associated with PD. Within this homogeneous cohort, two different groups were identified in relation to the genotype (patients carriers of GBA mutation and patients noncarriers of genes associated with PD) and the non-motor characteristics were compared in the two groups. We have therefore investigated the autonomic nervous system, through the study of cardiovascular reflexes and analysis of autonomic symptoms, and cognitive-behavioral functions in patients with PD associated with mutations in the GBA gene. The results were compared with the control group. The study showed that patients with PD associated with mutations in the GBA gene had higher frequency of sympathetic dysfunction, depression, anxiety, apathy, impulsivity, as well as disorders of impulse control compared to noncarriers patients. In conclusion, patients GBA positive can manifest a multidomain non-motor symptom profile with autonomic, cognitive and behavioral symptoms. Therefore, the therapeutic approach in these patients should include a thorough assessment of non-motor symptoms and their monitoring in the follow-up, in order to optimize the results and reduce the risk of complications.
24
PANETTA, PAOLA. "Alterazioni del gene NPM nelle mielodisplasie con delezione 5q-." Doctoral thesis, Università degli Studi di Roma "Tor Vergata", 2009. http://hdl.handle.net/2108/1124.
Full textAbstract:
Myelodysplastic syndromes (MDS) include a heterogeneous group of disease characterized by dysplasia of one or more bone marrow cell lineages, usually with prominent ineffective erythropoiesis and genomic instability leading to anaemia and enhanced risk to transformation to secondary acute myeloid leukemia (AML) . Thus MDS is often diagnosed on the basis of chronic macrocytic anaemia accompanied or not by leukocytopenia and/or thrombocytopenia. The deletion of 5q (5q-) is a frequent clonal chromosomal abnormality in patients with MDS. MDS with 5q- as a sole chromosome alteration is characterized by isolated anaemia, elevated platelet count and a favourable prognosis when compared to other forms of MDS . When the 5q- accompanies additional chromosome defects, it leads to poor-risk karyotypes with dramatically different prognostic features .
NPM1 is a versatile nuclear phosphoprotein that plays multiple roles in ribosome biogenesis and transport, cytoplasmic-nuclear trafficking, centrosome duplication and regulation of p53 .
The NPM1 gene is located in chromosome 5q35 and is involved in a number of human haematopoietic malignancies, such as promyelocytic leukaemia , anaplastic large cell lymphoma , and AML. NPM1 has also been found mutated in approximately 35% of acute myeloid leukaemia cases . Furthermore the 5q region to which NPM1map is deleted in a number of MDS and loss of chromosome 5 is a frequent finding in MDS . In a recent paper, Grisendi and co-workers showed that NPM1 is essential to maintain genomic stability. They demonstrated that NPM1 is haploinsufficient for regulating centrosome duplication as NPM1 heterozygous cells show aberrant centrosome numbers, genomic instability and aneuploidy. We analyzed the presence of NPM1 gene deletion, methylation and mutations in 45 patients affected by MDS and in 5 patients with AML secondary to MDS carrying the 5q- abnormality as sole chromosomal alteration or associated with additional chromosome defects.
Group 1 (17 patients) consisted of patients who present the 5q deletion as sole anomaly as determined by cytogenetics: 10 AR (59%), 1 ARS (6%), 1 AREB (6%), 4 AREB-T (23%) and 1 AML (6%). Group 2 ( 33 patients) consisted of patients who present the 5q deletion associated with additional chromosome defects: 5 AR (15%), 1 ARS (3%), 10 AREB (24%), 13 AREB-T (46%) e 4 AML (12%) The CpG island of the NPM1 gene was unmethylated in all the samples analyzed including patients with isolated 5q- and with complex karyotype. The mutational status of NPM1 exon 12 showed wild type NPM1 in all patient. The FISH analysis of the NPM1 locus revealed deletion of one copy of the gene in 7 cases. Interestingly all the cases with NPM1 deletion are always associated with complex karyotypes and a high-risk disease. May be considered that the aploinsufficienza of the NPM1 gene is not sufficient alone to determine the occurrence of a complex karyotype but may contribute with other genetic mechanisms for its establishment.
With the test of Fisher, has shown a trend of association between the deletion of NPM1 and complex karyotype (p = 0.08); most likely by increasing the number of cases analyzed could be obtained a statistical significance.
25
Etebari, Maryam <1983>. "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/7434/1/etebari_maryam_tesi.pdf.
Full textAbstract:
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.
26
Etebari, Maryam <1983>. "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/.
Full textAbstract:
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.
27
Mattiaccio, Alessandro <1985>. "Molecular characterization of gene defects associated with Progressive Familial Intrahepatic Cholestasis by Next Generation Sequencing." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2020. http://amsdottorato.unibo.it/9386/1/mattiaccio_alessandro_tesi.pdf.
Full textAbstract:
Progressive Familial Intrahepatic Cholestasis (PFIC) is a group of autosomal recessive diseases that affects especially newborns and children, with progression to liver failure in the first decades of life. PFIC is classified into five types based on the genetic defect involved in bile transport. It is caused by homozygous or compound heterozygous mutations in ATP8B1, ABCB11, ABCB4, TJP2 and NR1H4 genes. Other benign late-onset phenotypes and non-progressive forms (BRIC, LPAC, DIC and ICP) are caused by heterozygous mutations in the same gene pattern. Other genes have been recently involved in both progressive and non-progressive forms.
The aim of the project is to develop and validate a broad, reliable, rapid and cost-saving NGS genetic test for PFIC patients. 96 patients were tested for the first described genes and 80 patients were sequenced with the latest discovered candidate genes for PFIC and other related benign phenotypes. Bioinformatic and statistic pipelines were applied.
A total of 184 different variants has been identified in our cohort: 18 pathogenic, 46 VUS, 44 likely benign and 76 benign. P/LP mutations were found in 12% of patients: 2 in ATP8B1, 3 in ABCB11, ABCB4 and TJP2 each, one in ABCC2, JAG1, NOTCH2. Many patients had multiple variants in several genes. Patients had from 7 to 35 variants each and some SNPs were significantly associated with biochemical parameters and phenotypic features (e.g. liver fibrosis) that could better explain clinics and accelerate the progression to liver failure.
Our detection rate is according to other studies proposing multi-gene panels. Our analysis may be useful for the molecular diagnostics of PFIC and a better characterization and understanding of the linking between molecular defects and different subtypes of the disease. The high SNPs prevalence let us to hypothesize a synergistic haplotype effect in determining different multifactorial cholestasis phenotypes and overlapping features.
28
CAPO, VALENTINA. "Development of regulated lentiviral vectors for gene therapy of x-linked chronic granulomatous disease." Doctoral thesis, Università degli Studi di Roma "Tor Vergata", 2013. http://hdl.handle.net/2108/209901.
Full textAbstract:
Introduction. Chronic Granulomatous Disease (CGD) is
caused by defects of the NADPH oxidase complex, responsible
for the production of the oxidative burst in phagocytes.
Patients present increased susceptibility to life-threatening
fungal and bacterial infections and are treated with a lifelong
prophylaxis. Currently, the only curative option is bone
marrow transplantation. Gene therapy with hematopoietic
stem cells (HSC) may represent a valid alternative to
conventional transplant. Clinical trials for X-CGD employing
gp91phox-expressing gammaretroviral vectors have been limited
by insertional oncogenesis and lack of persistent engraftment.
Methods. We developed a novel strategy based on regulated,
self-inactivating lentiviral vectors (LVs) that target gp91phox
expression to the differentiated myeloid cells while sparing
HSC, to reduce the risk of genotoxicity and perturbation of
reactive oxygen species levels. Targeting was obtained by a
myeloid-specific promoter (MSP) and a posttranscriptional,
microRNA mediated regulation. We designed different
therapeutic gp91phox-expressing LVs for CGD gene therapy: 1)
PGK.gp91, in which gp91phox is driven by an ubiquitous cellular
promoter; 2) MSP.gp91, to control the transgene expression at
transcriptional level using a myeloid specific promoter; 3)
PGK.gp91_126T(2), in which we exploited the miRNA system,
incorporating miR-126 target sequences, to prevent the
transgene off-target expression in HSC; 4) MSP.gp91_126T(2),
combining the posttranscriptional de-targeting with the MSP.
Vectors were tested in human cell line, human bone marrow
HSC, their progeny differentiated in vitro and in the mouse
model of X-CGD.
Results. All vectors restored gp91phox expression and NADPH
oxidase function in human X-CGD PLB-985 cell line and in
myeloid cell lines and in macrophages from peripheral blood
monocytes of 3 X-CGD patients (22-48%). While unregulated
LVs ectopically expressed gp91phox in CD34+ cells, both
transcriptionally and post-transcriptionally regulated LVs
substantially reduced this off-target expression. By combining
transcriptional and posttranscriptional targeting in the dual
regulated vector, we achieved high levels of myeloid-specific
transgene expression, entirely sparing the most primitive
CD34+CD38-CD90+ HSC compartment (5-fold reduction). XCGD
mice transplanted with all vectors engrafted and restored
gp91phox expression, with 20-70% of granulocytes and
monocytes expressing human gp91phox. MSP-driven vectors
were superior in maintaining regulation during BM
development as well as in peripheral blood B and T cells.
Oxidase activity in corrected granulocytes was superior using
MSP-driven vectors (38-59% of WT NADPH oxidase activity)
as compared to PGK (17-23%). The MSP transcriptional control combined to miR-126 detargeting represent a
promising approach for further clinical development of
gp91phox therapeutic vectors.
29
RIGAMONTI, ALESSANDRA. "TRANSCRIPTOMIC ANALYSIS OF HUMAN CIRCULATING MONOCYTES: FOCUS ON MEMBRANE-SPANNING 4A GENE FAMILY MEMBERS." Doctoral thesis, Università degli Studi di Milano, 2021. http://hdl.handle.net/2434/844784.
Full textAbstract:
Within the mononuclear phagocyte system (MPS), monocytes represent the unique population able to operate as both effector and precursor cells. These incredibly plastic cells, mainly present in peripheral blood, are essential components of the innate immune system and play central roles both in homeostatic and pathological conditions. Monocytes are key determinants for the surveillance of endothelial integrity and repair, regulation of wound healing and replenishment of tissue resident macrophages (TRMs). Moreover, they serve as first line of defence against infections and hold a key position in several human diseases. Despite the fact that current classification distinguishes three major subsets (classical, intermediate and non-classical), monocytes represent an extremely heterogeneous population in terms of phenotype and specialized functions.
To overcome the remarkable lack of consensus on the identity and interrelationship of monocyte subsets, we have performed a single cell RNA sequencing analysis on circulating mononuclear cells of healthy donors. We identified 8 cluster of monocytes. C0 and c2 resembled neutrophil-like monocytes (NeuMo) and, together with c1, displayed distinct inflammatory programs and activation states. Of the two other clusters of classical monocytes, c7 significantly expressed high levels of antiviral genes, including IFN-related genes, while c12 corresponded to circulating monocyte-platelet aggregates (MPA). C6 and c3 resembled the intermediate and non-classical monocyte populations, respectively. Finally, a small cluster of CD16+ cells (c13) was characterized by the specific expression of genes of the complement system. We then moved to the analyses of a public transcriptomic dataset obtained from peripheral blood mononuclear cells (PBMCs) of gastrointestinal cancer patients at 3 time points of treatment. We were able to identify all previously defined monocyte subsets. Preliminary data showed the specific expansion of the IFN-related cluster c7 exclusively in responder patients after treatment with immunotherapy, suggesting a potential role of this population in response to therapy.
Analyses of the Membrane-Spanning 4-domain subfamily A (MS4A) protein family, representing a set of proteins whose roles in regulating myeloid cell function are now emerging, identified MS4A4A as potential markers for the commitment to circulating non-classical monocytes in humans. The expression of MS4A4A on a fraction of CD16+ monocytes was further confirmed at the protein level by flow cytometry. Given the selective expression of this protein on CD16+ cells and based on solid preliminary data from our laboratory, we
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hypothesized that MS4A4A may play a role in the biology of this subset of cells. Interestingly, bulk RNA sequencing of CD16-MS4A4A-, CD16+MS4A4A- and CD16+MS4A4A+ monocytes revealed enrichment of the Fc gamma receptor (FcγR) and Fc epsilon receptor (FcεR) pathways in MS4A4Apos cells. Implementation of these findings with functional assays is in program and will be essential to deeper investigate a possible role of MS4A4A as modulator of FcRs function in monocytes.
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Gilenmyr, Cheryl, and Kihl Charlie J. "Meningen med grammatik." Thesis, Högskolan i Borås, Akademin för bibliotek, information, pedagogik och IT, 2016. http://urn.kb.se/resolve?urn=urn:nbn:se:hb:diva-10302.
Full textAbstract:
Denna undersökning är en kvantitativ analys av elevtexters ordklassfördelning och ordklassers semantiska egenskaper.Det här arbetet ämnar undersöka eventuella samband mellan fördelningen av ordklasser och deras semantiska egenskaper i elevtexter av berättande karaktär och de olika betyg som de givna texterna erhåller.Dessutom undersöks dessa grammatiska variablers funktion för skrivandet i ett försök att lyfta och granska grammatikundervisningens roll inom den svenska skolan – och vidare diskutera och motivera hur grammatikundervisningen kan utformas för att höja funktionaliteten och relevansen, och därigenom förhoppningsvis kunna öka intresset för grammatik hos eleverna. Vi ämnar även utreda och ge en överblick över vad inom ordklasserna och deras semantik som kan vara värdefull kunskap för utformningen av undervisningen och för elevernas textskapande.Urvalet består av autentiska elevtexter hämtade ur nationella proven i svenska skrivna av årskurs nio läsåret 2013. Texterna har analyserats manuellt med ett kvantitativt stilistiskt angreppssätt.Resultatet påvisar några generella samband mellan en del olika variabler och de olika betygsnivåerna. En högre användningsfrekvens av adjektiv tenderar att ge ett högre betyg, så även för abstrakta substantiv. Högre betyg kan också förknippas med en högre användning av mentala och relationella processer – och därigenom har texterna på A-nivå även en högre andel statiska verb, då dessa ofta realiseras genom relationella processer. Texterna på A-nivå uppvisar också ett enhetligare tempusbruk jämfört med texterna på E-nivå, som istället tenderar att mer frekvent skifta tempus på ett oregelbundet sätt. Texterna på E-nivå upprepar gärna också samma ord i större utsträckning än de övriga betygsnivåerna. Undersökningen och resultatet kopplat till litteraturen och tidigare forskning motiverar en genrebaserad undervisning som ökar elevernas medvetenhet om olika texters uppbyggnad.
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Monti, L. "FUNCTIONAL STUDY OF MARK4, A GENE ENCODING FOR TWO PROTEIN ISOFORMS,IN GLIOMA AND NORMAL CELLS." Doctoral thesis, Università degli Studi di Milano, 2012. http://hdl.handle.net/2434/168374.
Full textAbstract:
MARK4 (MAP/Microtubule Affinity-Regulating Kinase 4) belongs to a family of serine-threonine kinases phosphorylating Microtubule Associated Proteins, causing their detachment from the microtubules (MTs) and thus increasing MTs dynamics. MARK proteins show high homology with PAR complex proteins family, involved in assessing cell polarity during embryogenesis, epithelial morphogenesis, neural differentiation, and cell migration. MARK proteins are thus implied in several processes involving MT network: cytoskeleton dynamics, cell polarity, centrosomes formation, chromosomal segregation, cytokinesis.
The MARK4 gene (19q13.2) encodes at least two alternatively spliced isoforms, L and S, differentially expressed in human tissues. MARK4S is the predominant isoform in normal brain and post-mitotic neurons. MARK4L has been found up-regulated in glioma cell lines and neural progenitors, as well as in hepatocarcinoma cell lines, suggesting a role in cell proliferation. The dual nature of MARK4 isoforms has been pinpointed by their expression profile in glioblastoma-derived cell lines and neural stem cells (NSCs), other than glioma, and the balance of the two isoforms, favouring the L splicing variant in glial tumors, is being investigated as a potential target of dysregulation in gliomagenesis. A linked view is whether the predominant expression of MARK4L, isoform of a gene involved in the microtubule dynamics, may concur to mitotic errors during gliomagenesis.
Both isoforms of MARK4 have been found associated to centrosomes and midbody, in glioma as well as in normal cells, suggesting that the kinase might have a role in all phases of the cell cycle. Moreover, MARK4L showed an additional nucleolar localization in glioma, raising the idea that, in tumors, the L variant has isoform specific functions and interactions with nucleolar components.
To verify the functional impact of MARK4 gene in glioma and in normal cells and to define the role of MARK4S and L isoforms with respect to their subcellular localization, we set up a functional study by RNA interference in glioblastoma (GBM) cell lines and normal fibroblasts, with specific silencing of both MARK4S and MARK4S+L.
We showed that MARK4 depletion determines heavy alterations in the cell shape of both G-32 GBM cell line and fibroblasts, corroborating MARK4 implication in cytoskeleton organization, and in accordance with the known role of MARK proteins in MTs dynamics.
MARK4 silencing particularly affected the centrosome cycle. Silenced G-32 GBM cell line and fibroblasts presented most of cells with the duplicated centrosome, apical to the nucleus, as typical of G1/S transition, while differently, control cells displayed centrosomes in all phases of the centrosome cycle. Accordingly, after MARK4 silencing, cell cycle analysis showed in both tumor cells and fibroblasts, an increase of G1 cells fraction and a strong reduction of mitoses, most of which displayed aberrations of spindle poles. These findings indicate that MARK4 depletion targets the G1/S transition checkpoint, probably knocking down a positive regulator.
It is worth noticing that the observed alterations of cell morphology, centrosome and cell cycle progression, and of mitosis were more pronounced when silencing MARK4S+L, as if the depletion of the sole MARKS might be partially compensated by MARK4L, being both isoforms localized at centrosomes and midbody.
MARK4 silencing on nucleoli revealed that the L isoform of MARK4 is not a specific marker of tumor (glioma) cell lines, in that it was also detectable in fibroblasts. However, MARK4 depletion showed a nucleolar pattern different in G-32 GBM cells from that of fibroblasts: G-32 showed, after both anti-MARK4S and MARK4S+L siRNA, a pronounced intensity of MARK4L signal in several nucleoli, while some others appeared silenced. In contrast, silenced fibroblasts showed, particularly after anti-MARK4S+L siRNA, several nucleoli unlabelled with MARK4L. It’s currently unknown the molecular basis for this difference which may be more than a quantitative one due to the higher levels of MARK4L in the nucleolar compartment of tumor cells where it’s generally upregulated.
The concomitant alterations of both centrosomal and nucleolar compartments highlighted by MARK4 silencing raise the hypothesis of a MARK4 role in the regulation of the Nucleus - (Nucleolus) - Centrosome (NC) axis. NC - axis is oriented and paired with the polarization and migration axis of many cell types, including fibroblasts and neural cells. Microtubules act as major actors in organizing a dynamic structure along NC - axis, regulating nuclear movement during cell migration and polarization. Since MARK4 cooperates with the microtubule network, its depletion may interfere with the correct orientation of the NC – axis.
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CHEN, XUE FEN. "FUNCTIONAL AND MECHANISTIC ANALYSES OF HISTONE DEACETYLASES (HDAC3) IN INFLAMMATORY GENE CONTROL." Doctoral thesis, Università degli Studi di Milano, 2011. http://hdl.handle.net/2434/155505.
Full textAbstract:
In this study, we investigated the role of individual class I histone deacetylases (HDACs) namely Hdac1, -2 and -3 using retroviral RNA interference to define their specific contribution to control an inducible gene expression program, namely inflammatory gene expression in 3T3 fibroblasts and primary macrophages. In addition to genes showing the expected transcriptional de-repression, we observed 8/20 genes in a test set being down-regulated following individual HDAC depletion. The requirement for HDACs function in gene induction as opposed to the more commonly observed role as transcriptional repressors may either underlie an indirect consequence of impaired HDAC mediated repression or a direct involvement of Hdac1 and Hdac3 in inducible gene activity. Therefore, we extended both the in vitro and in vivo analyses using conditional knockout (KO) mice. Genetic deletion of Hdac3 indicates that Hdac3 is required for the activation of 45% of the lipopolysaccharides (LPS)-induced genes. Global analysis of histone H4 acetylation showed that transcriptional down-regulation in Hdac3-/- cells did not correlate with increased histone acetylation, suggesting the possible involvement of indirect or secondary effects. We found that the LPS-inducible, Hdac3-dependent genes include a large group of interferon-β (IFNβ)-inducible genes (eg. IP10, Irf1) and another group (eg. Il-6) that may be regulated by AP-1 family proteins. In addition, gene expression analyses identified interferon-signalling pathway as being impaired in Hdac3-/- cells. Basal and inducible Ifnβ transcriptions require cJun/AP-1 and the decreased amount of AP-1 family proteins in Hdac3-/- cells may explain the lack of Ifnβ activation and the increased acetylation in genomic regions.
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Bacciaglia, Alessandro. "Caratterizzazione degli elementi genetici che veicolano il gene erm(B) in Streptococcus pyogenes." Doctoral thesis, Università Politecnica delle Marche, 2008. http://hdl.handle.net/11566/242461.
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Wessels, Elsabet. "Ontwikkeling van ’n koringkwekery met gestapelde, spesie-verhaalde roesweerstand." Thesis, Stellenbosch : University of Stellenbosch, 2010. http://hdl.handle.net/10019.1/5459.
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Thesis (MSc (Genetics))--University of Stellenbosch, 2010.Includes bibliography.
ENGLISH ABSTRACT: Wheat rust is a significant contributor to the total impact of diseases on sustainable wheat production. Genetic resistance, produced by using resistance genes from wheat and other related wild species, is the simplest and most cost-effective way to guard against these diseases. The pyramiding of resistance genes in a single line is a vital practice in bringing about durable resistance. This study aimed to develop a series of doubled haploid (DH) wheat lines containing combination's of wild species genes for rust resistance. Rust resistance genes Lr19 (7BL), Sr31/Lr26/Yr9/Pm8 (1BS) and Lr54/Yr37 (2DL) were combined by means of crossing. Breeders. lines which have complex resistance including Lr24/Sr24 (3DL), Lr34/Yr18 (7D), Sr36 (2BS) and Sr2 (3BS), were used. Marker assisted selection (MAS) was used to type populations for the above mentioned genes. Using the DH method (maize pollination technique), an inbred population was developed from the selected lines, after which the lines were characterised molecularly for the resistance gene translocations which they contain. The study produced 27 lines with diverse genetic profiles. Seven lines contain four translocations (Lr24/Sr24, Lr34/Yr18, Sr2 and Lr19 or Sr31) each, 11 lines contain three genes each, six lines contain two genes each and only three lines contain a single translocation (Lr24/Sr24). The reality that rust pathogens have already overcome three of the resistance genes in the final population . Lr19, Sr31 and Sr24 . is a clear indication of the value of using non-major gene resistance for bringing about durable resistance. The focus should fall ever more greatly upon the application of quantitative trait loci (QTL) for this purpose, which will result in MAS contributing to the development of more durable resistance. The value of the integration of MAS and DH in combination with conventional breeding practices in breeding programmes has already been illustrated internationally for increasing the rate of cultivar development and this is reaffirmed by this study.
AFRIKAANSE OPSOMMING: Koringroes lewer jaarliks .n beduidende bydrae tot die totale impak van siektes wat volhoubare koringverbouing belemmer. Die mees eenvoudige en koste-effektiewe verweer teen hierdie siektes is genetiese weerstand, wat deur weerstandsgene vanaf koring, sowel as wilde verwante spesies, bewerkstellig word. Die stapeling van weerstandsgene in .n enkele lyn word as .n onontbeerlike praktyk om duursame weerstand tot stand te bring, geag. Hierdie studie het ten doel gehad om .n reeks verdubbelde haploiede (VH) koringlyne te ontwikkel wat kombinasies van wilde spesie gene vir roesweerstand bevat. Roesweerstandsgene Lr19 (7BL), Sr31/Lr26/Yr9/Pm8 (1BS) en Lr54/Yr37 (2DL) is deur middel van kruisings gekombineer. Telerslyne wat oor komplekse weerstand beskik wat Lr24/Sr24 (3DL), Lr34/Yr18 (7D), Sr36 (2BS) en Sr2 (3BS) insluit, is gebruik. Merker-bemiddelde seleksie (MBS) is gebruik om populasies vir bogenoemde gene te tipeer. .n Ingeteelde populasie is vanaf die geselekteerde lyne met behulp van die VH metode (mielie-bestuiwing tegniek) ontwikkel, waarna die lyne molekuler vir die weerstandsgeentranslokasies waaroor hul beskik, gekarakteriseer is. Die studie het 27 lyne met diverse genetiese profiele opgelewer. Sewe lyne bevat vier weerstandsgeentranslokasies (Lr24/Sr24, Lr34/Yr18, Sr2 en Lr19 of Sr31) elk, 11 lyne beskik oor kombinasies van drie gene elk, ses bevat twee gene elk en slegs drie lyne beskik oor .n enkele translokasie (Lr24/Sr24). Die realiteit dat die roespatogene reeds drie van die weerstandsgene in die finale populasie . Lr19, Sr31 en Sr24 . oorkom het, benadruk die waarde van die gebruik van nie-hoofgeenweerstand vir die daarstelling van duursame weerstand. Die fokus behoort toenemend meer op die aanwending van kwantitatiewe kenmerk-loci (QTL) vir hierdie doel te val en sal sodoende teweegbring dat MBS bydra tot die ontwikkeling van meer duursame weerstand. Die waarde van die integrasie van MBS en VH in kombinasie met konvensionele telingsmetodiek is reeds internasionaal vir die versnelling van kultivarontwikkeling aangetoon en word ook deur hierdie studie herbevestig.
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BRUNO, GEMMA. "LIVER FIBROSIS IMPAIRS HEPATOCYTE TRANSDUCTION BY AAV VECTORS." Doctoral thesis, Università degli Studi di Milano, 2023. https://hdl.handle.net/2434/955885.
Full textAbstract:
Adeno-associated viral vectors (AAVs) are the most promising tools for liver directed gene therapy. However, integrity of hepatic architecture has been
considered pre-requisite for efficient gene delivery and clinical studies have been
addressed toward patients with no or negligible hepatic damage and fibrosis.
Preliminary evidence suggests that AAV-mediated gene transfer to hepatocytes
may be hampered by liver fibrosis, but knowledge about AAV vector interactions
with fibrotic livers is still very limited. In the present study, we investigated
hepatocyte transduction and biodistribution of AAV8-based vectors, commonly used
in liver-directed gene therapy clinical trials, in the context of liver fibrosis.
Analysis of three mouse models of induced and genetic liver fibrosis revealed that
fibrotic livers are transduced less efficiently by AAV8 and this results from reduced
vector uptake by the liver. Moreover, liver fibrosis altered blood vector clearance
and vector biodistribution in extra-hepatic organs.
Overall, these findings demonstrated that liver fibrosis impairs AAV-mediated gene
transfer to hepatocytes and highlight the relevance of the limitations posed by liver
fibrosis to efficient and safe gene transfer.
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Pierantonelli, Irene. "Ruolo del pancreatic duodenal homeobox gene protein 1 nella modulazione della risposta al danno delle cellule epatiche." Doctoral thesis, Università Politecnica delle Marche, 2013. http://hdl.handle.net/11566/242566.
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Rende, Francesca. "Kinetics and regulation of HTLV-1 gene expression." Doctoral thesis, Università degli studi di Padova, 2011. http://hdl.handle.net/11577/3421976.
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ABSTRACT
Human T-Lymphotropic virus type 1 (HTLV-1) is the causative agent of two distinct pathologies, adult T-cell leukemia/lymphoma (ATLL), an aggressive malignancy of mature CD4+ T-cells, and tropical spastic paraparesis/HTLV-1-associated myelopathy (TSP/HAM), a demyelinating neurodegenerative disease.
The HTLV-1 expression strategy is characterized by the production of plus- and minus-strand transcripts, alternative splicing and polycistronic translation. This strategy greatly increases the coding potential of the virus, resulting in expression of several regulatory and accessory genes (Tax, Rex, p12, p13, p21rex, p30tof and HBZ) in addition to the structural proteins and virion-associated enzymes common to all retroviruses (Gag, Pro, Pol and Env).
In spite of over 30 years of studies, several key features of the HTLV-1 life cycle and pathogenicity remain obscure. In particular, it is still unclear whether HTLV-1 gene expression is characterized by latency patterns, whether the different viral genes follow distinct kinetics of expression and, if this is the case, which molecular mechanisms control these phenomena.
The work described in the present thesis was aimed at understanding these aspects of HTLV-1 regulation. To this end we optimized a Real Time RT-PCR method using splice-site-specific primers to quantitate the different HTLV-1 transcripts and their kinetics of expression in peripheral blood mononuclear cells (PBMCs) isolated from HTLV-1-infected individuals and in cells transfected with HTLV-1 molecular clones. Results indicated that expression of HTLV-1 mRNAs follows a distinct timing upon reactivation of viral expression, with the mRNA coding for the Tax and Rex regulatory proteins acting as an early "master" transcript preceding expression of the other viral transcripts.
Although it is commonly accepted that Rex acts at a post-transcriptional level controlling the nuclear export and stability of viral mRNAs coding for the virion-associated proteins, the Rex-dependency of tax/rex, p12, p13, p21rex, p30tof and hbz transcripts has not been investigated so far. To test if the kinetics of HTLV-1 gene expression might be dependent on Rex function and to determine the Rex-dependence of individual HTLV-1 mRNAs, we generated a Rex knock-out HTLV-1 molecular clone and analyzed the nucleo-cytoplasmic compartmentalization of the viral mRNAs. Results demonstrated the strict Rex-dependency of the “two-phase” kinetics and revealed strong nuclear retention of hbz mRNAs, supporting their function as non-coding transcripts. Furthermore our results revealed that the Rex-responsiveness of the different HTLV-1 mRNAs is determined by a novel 72-nucleotides cis-acting regulatory sequence located upstream of exon 3.
Mathematical modelling underscored the importance of a temporal delay between the Tax and Rex functions, which was supported by experimental evidence of a delayed accumulation and longer half-life of Rex compared to Tax.
These data provide evidence for a temporal pattern of HTLV-1 gene expression, reveal major differences in the intracellular compartmentalization of HTLV-1 transcripts and, importantly, provide clues to a long-standing paradox of HTLV-1 regulation, i.e. the different Rex-dependence of viral transcripts in spite of the presence of the Rex-responsive element (RxRE) in the 3' untranslated region of all viral mRNAs.RIASSUNTO Il virus T-linfotropico umano di tipo 1 (HTLV-1) è l’agente eziologico di due distinte patologie, la leucemia/linfoma a cellule T dell’adulto (ATLL, adult T-cell leukemia/lymphoma), un'aggressiva neoplasia a carico dei linfociti T CD4+ maturi, e della paraparesi spastica tropicale/mielopatia associata ad HTLV-1 (TSP/HAM, tropical spastic paraparesis/HTLV-1-associated myelopathy), una patologia degenerativa del sistema nervoso centrale. La strategia di espressione genica di HTLV-1, caratterizzata dalla produzione di trascritti a partire da promotori localizzati sia nel filamento positivo che in quello negativo del genoma virale, da splicing alternativo e da traduzione bicistronica, incrementa notevolmente la capacità codificante di HTLV-1, con la conseguente espressione di numerosi geni regolatori ed accessori (Tax, Rex, p12, p13, p21rex, p30tof e HBZ) in aggiunta alle proteine strutturali e agli enzimi associati al virione, comuni a tutti i retrovirus (Gag, Pro, Pol ed Env). Nonostante oltre 30 anni di studi, diversi aspetti chiave del ciclo vitale di HTLV-1 e della sua patogenicità rimangono tutt'oggi non noti. In particolare, non è ancora chiaro se l'espressione genica di HTLV-1 sia caratterizzata da stadi di latenza, se i diversi geni virali presentino cinetiche di espressione distinte e quali meccanismi molecolari possano controllare questi fenomeni. Gli studi descritti nella presente tesi sono stati mirati a comprendere questi aspetti della regolazione genica di HTLV-1. A questo scopo abbiamo sviluppato un protocollo di Real Time RT-PCR associato all'impiego di primer specifici per le diverse giunzioni di splicing al fine di quantificare i diversi trascritti codificati da HTLV-1 e di analizzarne le cinetiche di espressione sia in cellule mononucleate di sangue periferico isolate da individui infettati con HTLV-1, che in cellule trasfettate con cloni molecolari di HTLV-1. I risultati ottenuti indicano che l'espressione degli mRNA codificati da HTLV-1 segue una precisa cinetica dopo riattivazione dell'espressione virale: l'mRNA codificante le proteine regolatrici Tax e Rex agisce come trascritto precoce che precede l'espressione degli altri geni virali. Sebbene sia comunemente accettato che Rex eserciti la sua funzione a livello post-trascrizionale controllando l'esporto nucleare e la stabilità degli mRNA che codificano le proteine associate al virione, fino ad oggi non è mai stata investigata la Rex-dipendenza dei trascritti p12, p13, p21rex, p30tof e hbz. Al fine di testare se le cinetiche di espressione genica di HTLV-1 osservate potessero dipendere dalla funzione di Rex e al fine di determinare la Rex-dipendenza dei singoli mRNA virali, abbiamo generato un clone molecolare di HTLV-1 knock-out per Rex e analizzato la compartimentalizzazione nucleo-citoplasmatica dei trascritti virali. I risultati ottenuti hanno dimostrato la stretta Rex-dipendenza delle cinetiche di espressione a "due fasi" ed hanno rivelato una forte ritenzione nucleare degli mRNA codificanti HBZ, supportando la loro funzione come trascritti non codificanti. Inoltre, i risultati ottenuti hanno dimostrato che la responsività a Rex dei differenti mRNA virali potrebbe essere determinata dalla presenza di una sequenza regolatoria di 72 nucleotidi che agisce in cis, localizzata a monte dell'esone 3. Infine, analisi matematiche hanno sottolineato l'importanza di un ritardo temporale tra le funzioni di Tax e di Rex, supportata dall'evidenza sperimentale di un ritardo nell'accumulo e di un'emivita più prolungata di Rex rispetto a Tax. I dati ottenuti in questo studio forniscono l'evidenza di una regolazione temporale dell'espressione genica di HTLV-1, rivelano una differente compartimentalizzazione degli mRNA virali e offrono una possibile spiegazione di un paradosso ancora irrisolto della regolazione di HTLV-1, ovvero la differente Rex-dipendenza dei trascritti virali, nonostante la presenza della sequenza responsiva a Rex (RxRE, Rex-responsive element) nella regione 3' non tradotta di tutti i trascritti virali.
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Corradini, Fabio. "Caratterizzazione molecolare delle mutazioni nei geni BRCA." Doctoral thesis, Università Politecnica delle Marche, 2009. http://hdl.handle.net/11566/242012.
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GHEZZI, DANIELE. "Identification and characterization of nuclear genes responsible for human mitochondrial disorders: fastkd2, responsible for a neurological disease associated with cox defiency and sdhaf1, encoding a complex II assembly, mutated in SDH-defective leukoencephalopaty." Doctoral thesis, Università degli Studi di Milano-Bicocca, 2009. http://hdl.handle.net/10281/7657.
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My researches during the DIMET project have been focused on the discovery of new genes responsible for mitochondrial disorders and the characterization of their role.
Recent epidemiological studies show that mitochondrial disorders have an incidence of 1:5000. These disorders are very heterogeneous and hence the diagnosis is difficult. Moreover mitochondrial dysfunctions are now clearly related to a wide range of disease conditions (i.e. neurodegeneration and cancer).
The majority of the inherited mitochondrial disorders, especially those with onset in infancy or childhood, are due to nuclear genes encoding proteins targeted to mitochondria. While identification of mutations in mitochondrial DNA has become relatively easy thank to the feasibility to perform the complete sequence analysis of mtDNA, the analysis of genomic DNA is more complicate and therefore the number of nuclear genes associated with mitochondrial diseases is still small.
Genome-wide analysis in families with autosomal recessive mitochondrial disorders could help to identify a genomic region to be further investigated. However, about one half/one third of the components of the mitochondrial proteome have yet to be identified, and this lack of information makes the search of candidate genes more difficult.
By linkage analysis or homozygosity mapping and prioritization of candidate genes, I studied subjects from multiconsanguineos families characterized by clinical pictures compatible with mitochondrial disorders.
In chapter 2, there is the report regarding the discovery of a nonsense mutation in two brothers displaying asymmetric brain atrophy, psychomotor regression and severe complex IV deficiency. The mutated gene codes for a mitochondrial predicted kinase that may have a role in apoptosis.
Using the same procedure, I take part in a project, which leads to the identification of the first assembly factor for complex II of the OXPHOS system (Chapter 3). Two different mutations were found in two pedigrees, with affected children characterized by acute psychomotor regression followed by spastic quadriparesis and/or dystonia. The pathogenic role of the mutations was confirmed in cellular and yeast models.
Finally, in chapter 4, there is the characterization of a protein, MR-1, already known and responsible for a movement disorder (PNKD, Paroxysmal non kinesigenic Dyskinesia). The mutant isoforms were erroneously localized into cytosol or membranes, whereas I demonstrated that they are mitochondrial and that the mutations reported so far in PNKD patients (and a new mutation identify in our study) are in the mitochondrial targeting signal (MTS). Hence PNKD could be considered a mitochondrial disease, due to a novel mechanism based on a deleterious action of the MTS.
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Wingqvist, Matilda. "Att undervisa om berättande texter : Hur man kan arbeta med genrepedagogik med fokus på berättande texter." Thesis, Karlstads universitet, Institutionen för språk, litteratur och interkultur, 2017. http://urn.kb.se/resolve?urn=urn:nbn:se:kau:diva-55167.
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The intention of my research is to increase understanding of what teaching with genre pedagogy, with focus on narrative texts, can look like. Through previous research and literature, I describe the emergence of genre pedagogy, why genre pedagogy is used, what a narrative text is and how it can be evaluated. The empirical material of the study consists of qualitative research interviews with teachers, as well as document analyses of the teachers’ educational plans with the purpose to gain a broader view of teaching with narrative texts. I have deliberately chosen the school I carried out my survey on. From my own experience, I know that this school work with genre pedagogy, which made it easier for me and the teachers as they knew the concepts, methods and terminology behind my questions. The results of the survey show that genre pedagogy promotes students’ learning, regardless of whether teachers choose to follow the structured circle model or focus on the creative personal writing. Both former researches and my teacher informants believe that the genre pedagogical teaching helps the students’ to develop subject language and their strategies for reading and writing texts in different genres. Nevertheless, as the results display, extra support is sometimes needed to further enhance students’ writing performance. The result also shows that the assessment of narrative texts usually has the best effect if it is written and formative.Intentionen med min undersökning är att öka förståelsen för hur undervisning med genrepedagogik med fokus på berättande texter kan se ut. Genom att utgå från tidigare forskning och litteratur beskriver jag bland annat genrepedagogikens uppkomst, varför genrepedagogik används, vad den berättande texten är, samt hur denna kan bedömas. Undersökningens empiri består av kvalitativa forskningsintervjuer gjorda med lärare, samt dokumentanalyser av lärarnas pedagogiska planeringar med syfte att få en bredare bild av undervisning av berättande texter. Skolan jag valt att utföra min undersökning på har jag medvetet valt då jag sedan tidigare vetat om att de arbetar genrepedagogiskt, vilket underlättade både för mig och lärarinformanterna då de kände till begreppen i mina frågeställningar. Resultaten av undersökningen visar att genrepedagogisk undervisning främjar elevernas lärande, oavsett om lärarna väljer att följa den strukturerade cirkelmodellen eller om de fokuserar på det kreativa personliga skrivandet. Både tidigare forskare och mina lärarinformanter menar att den genrepedagogiska undervisningen gör att både elevernas ämnesspråk och strategier att läsa och skriva texter inom olika genrer utvecklas. Trots detta framgår dock i resultatet att extra stöttning ibland behövs för att stärka elevernas skrivutveckling ytterligare. I resultatet framgår även att bedömningen av berättande texter oftast har bäst effekt om den är skriftlig och formativ.
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Dow, Michael Rhys. "The cloning and characterization of the mel-26 gene of Caenorhabditis elegans." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1998. http://www.collectionscanada.ca/obj/s4/f2/dsk2/ftp02/NQ34669.pdf.
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Spetz, Anna-Clara. "Vasomotor symptoms in men and the role of calcitonin gene-related peptide /." Linköping : Univ, 2002. http://www.bibl.liu.se/liupubl/disp/disp2002/med758s.pdf.
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Ponzo, Marisa Grace 1980. "Gene expression profiling of Met receptor tyrosine kinase-induced mouse mammary tumors." Thesis, McGill University, 2009. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=115881.
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Breast cancer is a heterogeneous disease comprised of distinct biological entities that correlate with diverse clinical outcomes. Gene expression profiling has divided this heterogeneity into luminal, ERBB2+ and basal molecular subtypes. Basal breast cancers are difficult to treat as they lack expression of candidates suitable for targeted therapies and are associated with poor outcome.Elevated protein level of the hepatocyte growth factor receptor, MET, is observed in 20% of human breast cancers and correlates with poor prognosis. However, the role of MET in mammary tumorigenesis is poorly understood. To address this, we generated a murine model that expresses weakly oncogenic mutants of Met (Metmt) in the mammary epithelium under the transcriptional control of the mouse mammary tumor virus promoter. We demonstrate that Metmt induces mammary carcinomas with diverse phenotypes and used gene expression microarrays to elucidate gene expression changes induced by Met. Since mammary tumors contained variable contents of epithelium and stroma, we used laser capture microdissection to procure epithelial cells for microarray analysis. Based on immunohistochemistry and expression profiling, we show that Metmt produces tumors with luminal or basal characteristics. From hierarchical clustering, Metmt-induced basal tumors clustered with murine models that share features of epithelial to mesenchymal transition and human basal breast cancers. Moreover, Metmt basal tumors clustered with human basal breast cancer. The status of MET among the human breast cancer subtypes has not previously been addressed. We demonstrate that MET levels are variable across molecular subtypes but show elevation in the basal subtype and correlates with poor outcome. We used a candidate gene approach derived from microarray data to gain an understanding of signals required for Met-dependent tumorigenesis. We investigated Nck adaptor proteins and demonstrate a role for Nck in cell motility and actin dynamics of Met-dependent breast carcinoma cells and show elevated expression in human basal breast cancers. By generating a unique mouse model in which Met is expressed in mammary epithelia, with the examination of MET levels in human breast cancer, we have established a novel link between MET and basal breast cancer. This work identifies poor outcome basal breast cancers that may benefit from anti-MET therapies.
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AMINI, NIA SHADI. "Investigation of effects of lymphoma associated gene polymorphisms and Aryl Hydrocarbon Receptor(AhR) activation on DNA." Doctoral thesis, Università degli Studi di Cagliari, 2019. http://hdl.handle.net/11584/270318.
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Abstract
Aim:To investigate whether AhR activation induces DNA damage, whether polymorphisms in genes related to risk of Non-Hodgkin lymphoma are associated with DNA damage, and whether the two conditions do interact with each other.
Material and Methods. Our study population included 36 subjects, randomly selected among the population controls participating in a case-control study on lymphoma in Sardinia, Italy, who donated a blood sample.We investigated 47 single nucleotide polymorphisms (SNPs) previously reported to convey risk of lymphoma; the Dual-Glo® Luciferase Assay System to detect activation of the aryl hydrocarbon receptor (AhR) by the serum of study subjects; and the COMET Assay to detect DNA damage.
Results:Activation of the aryl hydrocarbon receptor did not increase DNA damage in our study population. On the other hand, the mutant allele (G) of rs1056932/BCL6 increased the occurrence of DNA damage (p=0.045); such association was confirmed only among AhR negative subjects (p=0.025).
Conclusion: We observed excess DNA damage associated with a gene polymorphism, namelyrs1056932/ BCL6,previously reported in association with risk of lymphoma.No increase in DNA damage was associated with AhR activation per se, nor with the other gene polymorphisms we investigated.
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Trentin, Luca. "Microarray Analysis: a Leading Tool in the Classification and Biological Characterization of Pediatric Onco-Hematological Diseases." Doctoral thesis, Università degli studi di Padova, 2010. http://hdl.handle.net/11577/3427012.
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Gene Expression Profile (GEP) analysis through microarrays represents a powerful tool for the classification, the prediction and the identification of several leukemia subclasses. In this thesis, we have reported the results we have obtained applying the microarray technology to the study of pediatric onco-hematological diseases.
A huge amount of reports have highlighted the robustness of GEP in the classification of leukemia in both children and adults and these results support the application of microarrays in future routine diagnostic settings.
Since the quality of RNA used for the experiments is one of the critical factors when performing microarrays analysis and seeing that all laboratories commonly use their own distinctive RNA isolation protocol, we have questioned the influence of the three most frequently used extraction protocols in the gene expression profile analysis of pediatric leukemia. Our data have showed that different sample preparation procedures do not impair samples classification and that the underlying biological characteristics of the pediatric acute leukemia classes largely exceed the variations between different RNA preparation protocols.
We have then applied GEP analysis to the study of MLL/AF4-rearranged B-cell precursor acute lymphoblastic leukemia (BCP-ALL). Among the MLL/AF4 BCPs leukemia samples, we have identified the presence of two subgroups of patients characterized by a distinctive gene expression signature in which the down-regulation of the HOXA genes is a particularly outstanding factor. HOXA genes deregulation, indeed, is commonly believed to be a key mechanism of MLL-fusion gene mediated leukemogenesis. Apart from the differential HOXA genes expression level in these subgroups of patients, no transcriptional deregulation of other known MLL-related genes (i.e. MENIN, HOXC8 and MEIS1) could be identified.
We have also performed a microRNA expression profile analysis of the patients characterized by the up- or down-regulation of HOXA genes and we have demonstrated that they are characterized also by a distinct microRNA signature. Interestingly, patients displaying a low expression value of HOXA genes do not express the miR-196b which is located within the HOXA cluster and which is involved in leukemogenesis.
Furthermore, we have used microarray analysis to study Juvenile Myelomonocytic Leukemia (JMML). Remarkably, we could distinguish two distinct subgroups among the analyzed patients and this subdivision resulted to have a high prognostic value in the identification of subgroups of patients with distinct clinical outcome. The same result is not reproducible if the usual clinical factors (fetal Hb, age at diagnosis and platelet count) are applied.
Finally, we have focused on the role of the suppressor of cytokine signaling 2 (SOCS-2). This gene is reported to be up-regulated in stem cells and we have identified SOCS-2 as one of the most up-regulated genes in MLL/AF4 patients irrespective of HOXA gene expression level, when comparing t(4;11) samples with normal bone marrow controls. Transient silencing of SOCS-2 in the lymphoid cell line RS4;11 showed that SOCS-2 depletion induces apoptosis in silenced cells and that this process is characterized by the concurrent increased expression of TP53 and BAX. Thus, SOCS-2 up-regulation seems to be a mechanism in RS4;11 cells to impair apoptosis activation. We have analyzed SOCS-2 expression levels in patients belonging to several different ALL subclasses and have found that SOCS-2 is up-regulated in all but T-lineage leukemia. This finding suggests that SOCS-2 up-regulation could be a common mechanism in ALLs to prevent induction of apoptosis.L’analisi del profilo d’espressione genica mediante microarray rappresenta uno strumento utile per la classificazione delle leucemie in ambito diagnostico, l’identificazione di nuove sottoclassi di malattia e l’associazione di profili d’espressione genica con la prognosi. I molteplici lavori pubblicati nell’ambito delle malattie onco-ematologiche sia nell’adulto che nel bambino hanno evidenziato la robustezza della tecnologia microarray ed auspicano, quindi, l’ utilizzo dei microarrays in affiancamento alle metodiche “gold standard” per la diagnosi di leucemia. Considerando che la qualità dell’RNA di partenza è un fattore determinante per la buona riuscita di un esperimento di studio dell’espressione genica, abbiamo valutato se diverse metodiche di isolamento dell’RNA avessero una qualche influenza sulla variazione del profilo d’espressione genica. I risultati ottenuti nel nostro studio, analizzando diversi sottotipi di leucemie pediatriche, hanno evidenziato che le metodiche impiegate per l’estrazione dell’RNA non vanno ad influire sul profilo d’espressione genico e che quest’ultimo rimane, comunque, ben identificabile a prescindere dalla metodologia usata per l’isolamento dell’RNA. Applicando, poi, l’analisi microarrays alle leucemie a cellule precursori B e con traslocazione MLL/AF4, abbiamo individuato, all’interno di questo sottotipo di leucemia ritenuto fino ad ora omogeneo, due sottogruppi di pazienti caratterizzati da un differente profilo d’espressione genica in cui spiccava la diversa espressione dei geni HOXA. Questo risultato è alquanto sorprendente poiché la maggiore espressione dei geni HOXA è una caratteristica distintiva delle leucemie con riarrangiamento del gene MLL. Non abbiamo identificato nessuna altra variazione d’espressione di geni (per es. MENIN, HOXC8 e MEIS1) comunemente associati con le leucemie con riarrangiamento del gene MLL. Anche l’analisi del profilo dell’espressione dei microRNA ha dimostrato che questi pazienti possono essere suddivisi in due sottogruppi ben distinti ed, inoltre, ha evidenziato che i pazienti con bassa espressione dei geni HOXA non esprimono il microRNA mir-196b, che è localizzato nel medesimo cluster dei geni HOXA e che è coinvolto nei processi di leucemogenesi. Lo studio del profilo d’espressione genica di pazienti affetti da leucemia mielomonocitica giovanile (JMML) ci ha, poi, consentito di dividere i campioni analizzati in due sottogruppi. Questa suddivisione è associata, in modo altamente significativo, con la prognosi di malattia. Il medesimo risultato prognostico non è conseguibile prendendo in considerazione i fattori prognostici clinici standard (emoglobina fetale, età alla diagnosi e conta piastrinica). Infine, abbiamo studiato il ruolo del gene SOCS-2 nelle leucemie con riarrangiamento MLL/AF4. Questo gene, up-regolato nelle cellule staminali, è uno dei geni maggiormente espressi nei pazienti con MLL/AF4 rispetto ai controlli normali. Il silenziamento di SOCS-2 nelle cellule RS4;11 determina un aumento dell’apoptosi rispetto alle cellule silenziate con un siRNA di controllo ed una simultanea maggiore espressione di TP53 e BAX. L’over-espressione di SOCS-2 nelle cellule RS4;11 sembra essere, quindi, un meccanismo in grado di aumentare la sensibilità di queste cellule all’apoptosi. L’analisi dell’espressione di SOCS-2 in più pazienti affetti da leucemia linfoblastica acuta (LLA) ha evidenziato che SOCS-2 è over-espresso in tutte le LAL tranne le LAL a cellule T. Questo dato suggerisce che l’azione anti-apoptotica di SOCS-2 potrebbe essere comune in più sottotipi di leucemia.
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Mosca, A. "Calcium-Sensing Receptor: a candidate gene for Kidney Stone Disease." Doctoral thesis, Università degli Studi di Milano, 2007. http://hdl.handle.net/2434/43624.
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Colombo, E. "Significato clinico del poliformismo del gene NOD2/CARD15 nella storia chirurgica dei pazienti affetti da Morbo di Crohn." Doctoral thesis, Università degli Studi di Milano, 2007. http://hdl.handle.net/2434/64168.
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Davis, Ian. "Teaching Men: Masculinity, Narrative and Pedagogy." Thesis, Griffith University, 2014. http://hdl.handle.net/10072/367340.
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The Teaching Men project investigates masculinity, narrative and teaching by considering one central question; (How) are male teachers influenced by fictional narratives in the construction of masculinities within education? The exploration of this question is executed using three distinct yet corresponding research activities. Firstly by developing a methodological system of narrative analysis that is able to account for the influence of a fictional text alongside a reading of interview data. Secondly by focusing on a specific cohort of male teachers in order to measure the influence of a fictional text, illustrating possibilities of how masculinity can be enacted within education. Finally by assessing how the narrative nature of critical reflective practice enables the integration of fictional texts, and the literary tropes they contain, both widening and restricting perceptions of teachers and teaching. The work of the project demonstrates how fictional narratives and their encompassing ideologies can become a powerful force in the shaping of our professional identities, in this case as male teachers.
The Teaching Men project has two parallel research streams. The first stream focuses on a collection of 22 fictional narratives drawn from the teacher text genre. Each text describes the world of teachers and teaching from differing perspectives, in differing forms including, literary texts; dramatic works such as plays or musicals; feature films; and television and radio series. The teacher text genre is both popular and prolific; therefore three key criteria have been established to determine inclusion within the project.Thesis (PhD Doctorate)
Doctor of Philosophy (PhD)
School of Education and Professional Studies
Arts, Education and Law
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VALSECCHI, VERONICA. "Il ruolo del gene rat8/IFITM3 nel differenziamento in vitro delle cellule mammarie." Doctoral thesis, Università degli Studi di Milano, 2007. http://hdl.handle.net/2434/33614.
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In my laboratory we developed an experimental model system, based on two cellular clones, LA7 and 106 cells, that allow us to study rat mammary differentiation in vitro. The two cell lines differ in their ability to differentiate: the LA7, but not 106 cells, spontaneously or after exposure to differentiating agents, such as DMSO, are able to develop hemispheric structures called domes. The domes recapitulate, in vitro, the morphological and functional changes that occur, in vivo, in the mammary gland at pregnancy, when alveoli are formed During my PhD project, I demonstrated that the differentiation of LA7 cells, resulting in dome formation, requires the presence of a multimeric complex on the cell surface constituted by Ifitm3-Fyn-Shc1-alfa6beta1 integrin, and caveolin1. Each member of this complex plays an important role in dome formation as even the lack of one of these members affects the differentiation pathway. I can conclude that this protein complex at the surface of LA7 cells is necessary to initiate the signal transduction cascade that controls differentiation of mammary epithelial cells
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Lindeborg, Hanna. "Vad är ett riktigt slut? : en kritisk granskning av genrepedagogiken i arbetet med sagogenren." Thesis, Södertörns högskola, Lärarutbildningen, 2011. http://urn.kb.se/resolve?urn=urn:nbn:se:sh:diva-9757.
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With this paper the aim is to find out whether there are any differences in how five students from the north part of Africa structures their tales in comparison with the Nordic tale "The emperor‟s new clothes". The purpose is to look at how the genre pedagogy can be said to relate to these differences. What happens to the "deviant" structures? Based on literature and research on the subject, I have concluded that there are certain patterns of how a tale "should" be structured based on Western standards. These text structures are seen as typical for the genre and are therefore, in accordance with the subject plans, reproduced in the classroom. In the analysis of the student´s texts there has been shown and a rather large variation in how the tales are structured.