3 resultados para Nemo-Q
em AMS Tesi di Dottorato - Alm@DL - Università di Bologna
Resumo:
Anhidrotic Ectodermal Dysplasia (EDA), is the most frequent form among Ectodermal Dysplasias, hereditary genetic disorders causing ectodermal appendages defective development. Indeed, EDA is characterized by defective formation of hair follicles, sweat glands and teeth both in human patients and animals. EDA, the gene mutated in Anhidrotic Ectodermal Dysplasia, encodes Ectodysplasin, a TNF family member that activates NF-kB mediated transcription. This disease can occur with mutations in other EDA-NF-kB pathway members, as EDA receptor, EDAR and its adapter, EDARADD. Moreover, mutations in TRAF6, NEMO, IKB and NF-kBs genes are responsible for Immunodeficiency associated EDA (EDA-ID). Several molecules, as SHH, WNT/DKK, BMP and LTβ, have already been reported to be EDA pathway regulators or effectors although the knowledge of the full spectrum of EDA targets remains incomplete. During the first part of the research project a gene expression analysis was performed in primary keratinocytes from Wild-type and Tabby (EDA model mouse) mice to identify novel EDA target genes. Earlier expression profiling at various developmental time points in Tabby and Wild-type mouse skin reported genes differentially expressed in the two samples and, to increase the resolution to find genes whose expression may be restricted to epidermal cells, the study was extended to primary keratinocyte cultures established from E19 Wild-type and Tabby skin. Using microarrays bearing 44,000 gene probes, we found 385 “preliminary candidate” genes whose expression was significantly affected by Eda defect. By comparing expression profiles to those from Eda-A1 (where Eda-A1 is highly expressed) transgenic skin, we restricted the list to 38 “candidate EDA targets”, 14 of which were already known to be expressed in hair follicles or epidermis. This work confirmed expression changes for 3 selected genes, Tbx1, Bmp7, and Jag1, both in primary keratinocytes and in Wild-type and Tabby whole skin, by Q-PCR and Western blotting analyses. Thus, this study detected novel candidate pathways downstream of EDA. In the second part of the research project, plasmid constructs were produced and analyzed to create a transgenic mouse model for Immunodeficiency associated EDA disease (XL-EDA-ID). In particular, plasmids containing mouse Wild-type and mutated Nemo cDNA under K-17 epidermis-specific promoter control and a Flag tag, were prepared, on the way to confine transgene expression to mice epidermis and to determine EDA phenotype without immunodeficiency for a comparison to Tabby model phenotype. EDA-ID mutations reported in patients and selected for this study are: C417R (C409R in mouse), causing Zinc Finger protein domain destabilization and A288G (A282G in mouse) affecting oligomerization of the protein. Moreover, the ex-novo mutation, ZnF, C-terminal Zinc Finger domain deletion, was tested. Thus, the constructs were analyzed by transient transfection, Western blotting and luciferase assays techniques, detecting Nemo Wild-type and mutant protein products and residue NF-kB activity in presence of mutants, after TNF stimulation. In particular, MEF_Nemo-/- cell line was used to monitor NF-kB activity without endogenous Nemo gene. Results show reduced NF-kB activity in presence of mutated Nemo forms compared to Wild-type: 81% for A282G (A288G in human); 24% for C409R (C417R in human); 15% for ZnF. C409R mutation (C417R in human), reported in 6 EDA-ID human patients, was selected to prepare transgenic model mouse. Mice (white, FVP) born following K17-promoter-Flag-Nemo_C409R plasmid region pronuclear injection, were analyzed for the transgene presence in the genotype and a preliminar examination of their phenotype was performed. In particular, one mouse showed considerable coat defects if compared to Wild-type mice. This preliminar analysis suggests a possible influence of Nemo mutant over-expression in epidermis without immunodeficiency. Still, more microscopic studies to analyze hair subtypes, Guard, Awl and Zigzag (usually alterated inTabby mouse model), Immunohistochemistry experiments to detect epidermis restricted Nemo expression and sweat glands analysis, will follow. This and other transgene positive mice will be crossed with black mice C57BL6 to obtain at least two indipendent agouti lines to analyze. Theses mice will be used in EDA target genes detection through microarrays. Following, plasmid constructs containing other Nemo mutant forms (A282G and ZnF) might be studied by the same experimental approaches to prepare more transgenic model mice to compare to Nemo_C409R and Tabby mouse models.
Resumo:
The major index has been deeply studied from the early 1900s and recently has been generalized in different directions, such as the case of labeled forests and colored permutations. In this thesis we define new types of labelings for forests in which the labels are colored integers. We extend the definition of the flag-major index for these labelings and we present an analogue of well known major index hook length formulas. Finally, this study (which has just apparently a simple combinatoric nature) allows us to show a notion of duality for two particular families of groups obtained from the product G(r,n)×G(r,m).
Resumo:
The Mufarriḥ an-nafs (Soul-Cheerer), attributed to Badr ad-Dīn Muẓaffar Ibn Qāḍī Baʿlabakk, who served under the Ayyubids as the Chief Medical Officer of Damascus in the mid-13th century, was written as a comprehensive guide for physicians outlining different approaches to cheering the soul. The tractate is divided into ten chapters, which explore the nature of the soul, its distinction to the body as well as their connection through sensorial perception. Ibn Qāḍī Baʿlabakk distinguishes the bodily senses – hearing, vision, smell, taste, touch – and the inner senses, which he sees as stimulated through activities such as hunting and engagement in poetry and the sciences. The seventh chapter of the Mufarriḥ an-nafs includes an extended encyclopedia on materia medica as well as dispensatory of simple and compound drugs, which is devoted to treating the soul and remains unparalleled in the history of Islamicate medicine. My doctoral dissertation offers a complete recension and translation of the Mufarriḥ an-nafs based on a stemma codicum drawn from the seventeen extant text witnesses. The dissertation contextualizes the work, its author as well as sources, and features a text commentary that seeks to enable the reader to easily place and understand the Mufarriḥ an-nafs within the tradition of Galenic medicine. The glossaries on materia medica found at the end of the dissertation are aimed at facilitating access to the pharmacological dispensatory included in the seventh chapter.