Every year millions of individuals suffer from traumatic brain injury (TBI) leading to permanent disabilities and even death. Mild TBI (mTBI) is the most common form of TBI comprising about 80-90% of all occurrences. Following a CNS insult like an mTBI, astrocytes can undergo activation resulting in the transformation into reactive astrocytes (RAs). RAs also play an important role in brain remodeling following an mTBI. Research on the mechanical complexity of the brain has important implications for understanding brain function and dysfunction, as well as for the development of new diagnostic and therapeutic tools for neurological disorders. This study aimed to develop and utilize an emph{in vitro} mTBI platform to investigate the intricate mechanical interplay between the extracellular matrix (ECM) and astrocytes following a simulated mTBI. Cellular mechanisms underlying mTBI and the contribution of mechanical forces that result in prolonged brain damage are yet to be comprehensively understood. Successfully devised mechanical characterization techniques for tissue-engineered models were developed utilizing atomic force microscopy and rheology. Astrocyte exposure to high-rate overpressure revealed altered mechanical properties of the surrounding matrix and decreased expression of laminin and collagen IV, which are critical for brain function and may contribute to pathologies associated with mTBI. The developed platform and methods provide new insights into the mechanistic complexity underlying ECM-astrocyte interactions following an mTBI. / Master of Science / Every year, millions of people suffer from traumatic brain injury (TBI), which can lead to permanent disabilities or even death. The most common form of TBI is mild TBI (mTBI), which accounts for 80-90% of all cases. After a mTBI, astrocytes, the most common cell type in the brain, can become activated and turn into reactive astrocytes (RAs). RAs play an important role in the brain's recovery following a mTBI. Understanding the mechanical complexity of the brain is crucial for developing new diagnostic and therapeutic tools for neurological disorders. This study aimed to investigate the mechanical interplay between the modeled tissue and astrocytes following a simulated mTBI using an emph{in vitro} platform. Development of mechanical characterization techniques allowed for any alterations caused by the astrocytes to their environment to be detectable. The astrocyte exposure to the simulated mTBI revealed altered mechanical properties of the surrounding environment and decreased expression of proteins laminin and collagen IV, which are critical to brain function and may contribute to pathologies associated with mTBI. This study provides new insights into the mechanistic complexity underlying the interaction between astrocytes and their environment, which could lead to the development of new treatments.

The Drosophila heart is a tubular vessel surrounded by a dynamic scaffold of extracellular matrix (ECM) proteins. Heart development and function rely upon protease-mediated remodelling and turnover of the ECM, and changes in ECM composition correlate with age and cardiac disease. Previous research has shown that a family of proteases called matrix metalloproteinases (MMPs), and their inhibitors (TIMPs), are necessary for normal cardiac cell migration and lumenogenesis. The Drosophila heart expands considerably throughout growth, but the role of MMP activity has not been elucidated at this time. I examine the role of the two Drosophila MMPs, MMP1 and MMP2, as well as TIMP, in defining larval heart structure and ECM protein distribution. I observe heart phenotypes via immunofluorescence labelling and confocal microscopy using loss-of-function mutants, gene over-expression, and gene knock-down techniques. Reduced MMP1 function during embryogenesis correlates with myofibrillar disorganisation, whereas reduced MMP2 function or TIMP over-expression both result in cardia bifida as well as increased density and ectopic localisation of Collagen-IV and Pericardin. Post-embryonic MMP reduction compromises cardiac structural integrity but does not affect Pericardin localisation. Live imaging of the larval heart with optical coherence tomography (OCT) and light microscopy reveals that reduced MMP2 function correlates with decreased heart rate but not impaired dilation or contraction. These data suggest that MMP2 activity during embryogenesis is critical for larval heart development. In contrast, post-embryonic protease function appears to have a less pronounced effect on ECM protein distribution throughout larval development. / Thesis / Master of Science (MS) / The fruit fly (Drosophila) heart undergoes significant changes in organisation and size throughout development and growth. The heart is surrounded and supported by a network of extracellular matrix (ECM) proteins, which is regulated by proteases, including matrix metalloproteinases (MMPs). Previous research has shown that MMPs are required for normal heart formation. I demonstrate that a reduction in MMP activity during embryonic development results in larval heart defects and an increase in the disorganisation of ECM proteins around the heart, whereas reduction during larval development results in less pronounced protein mislocalisation. These findings are corroborated via over-expression of an MMP inhibitor.

Der pRB/E2F-Signalweg ist ein zentraler Regulator der Proliferationskontrolle in Säugerzellen, der in fast allen auftretenden Tumoren dereguliert ist. Durch unterschiedliche Mutationen in Komponenten dieses Signalwegs kommt es letzten Endes zu einer erhöhten Aktivität der E2F-Transkriptionsfaktoren und somit zu einer verstärkten Transkription von E2F-Zielgenen in diesen Tumoren. Um die molekularen Mechanismen der Rolle von E2F3 in der Zellzykluskontrolle und der Tumorigenese besser zu verstehen, wurden in dieser Arbeit per GST-Pulldown mit anschließender Massenspektrometrie neue potenzielle Interaktions-partner von E2F3 identifiziert. Ein identifizierter Interaktionspartner war die SNF2-ähnliche Helikase HELLS. HELLS interagiert in vitro und in vivo spezifisch mit der Marked Box-Domäne von E2F3, aber nicht mit anderen untersuchten E2F-Transkriptionsfaktoren, wie durch GST-Interaktionsstudien und Ko-Immunpräzipi-tationsexperimente demonstriert werden konnte. Durch Chromatin-Immunpräzipitation konnte zusätzlich gezeigt werden, dass E2F3 für die Rekrutierung von HELLS an E2F-regulierte Promotoren wie z. B. CDC6 oder p107 verantwortlich ist. Die shRNA-vermittelte Depletion von HELLS führte zu einer stark verminderten Induktion von allen untersuchten E2F-Zielgenen nach Serumstimulation und einem verspäteten Eintritt in die S-Phase der HELLS-depletierten Zellen, was zeigt, dass HELLS essenziell für die Induktion von E2F-Zielgenen ist. Bei der immunhistochemischen Untersuchung der E2F3- und HELLS-Expression in humanen Prostatakarzinomen zeigte sich, dass sowohl E2F3 als auch HELLS in späten aggressiven Stadien dieser Tumore sehr stark exprimiert sind, jedoch nur sehr schwach in den weniger aggressiven Tumoren. Diese Versuche zeigen, dass es sich bei HELLS um einen neuen Bestandteil des pRB/E2F-Signalwegs handelt, der eventuell in der Entstehung gewisser Tumorarten eine Rolle spielt und somit ein neues potenzielles Ziel für neuartige Krebstherapien darstellt. / The pRB/E2F pathway is a key regulator of proliferation in mammalian cells and is commonly mutated in human tumors. These mutations in the components of the pRB/E2F pathway lead to deregulated activity of the E2F transcription factors resulting in increased expression of E2F target genes. To further understand the molecular mechanisms of E2F3 in cell cycle control and its role in tumorigenesis new interaction partners for E2F3 were identified in the course of this thesis with the help of a GST-Pulldown approach coupled to mass spectrometric analysis. One of the identified interaction partners was the SNF2-like helicase HELLS. With the help of GST-interaction studies and Co-Immunoprecipitation assays it could be demonstrated that HELLS interacts specifically with E2F3 via its Marked Box domain but does not bind to the other investigated E2F transcription factors. HELLS could be detected at E2F target genes like p107 and CDC6 in vivo with the help of Chromatin-Immunoprecipitation assays. Furthermore, the forced recruitment of E2F3 to E2F target genes led to an enhanced binding of HELLS to these promotors suggesting that HELLS is recruited to E2F target genes via protein-protein interaction with E2F3. The shRNA-mediated depletion of HELLS led to a strongly reduced induction of E2F target genes and a delay in S-phase entry, showing that HELLS is essential for the induction of E2F target genes. During the immunohistochemical analysis of human prostate cancer specimens it became evident that both E2F3 and HELLS are strongly expressed in the more aggressive late stages but only weakly expressed in the early stages of this tumor type. These findings demonstrate that HELLS is a new component of the E2F/pRB pathway which might play a role in the development of certain tumors and might represent a new target for novel cancer therapies.

Distortion to hemodynamic, ischemic and metabolic stimuli during low load resistance exercise with blood flow restriction (BFR) may influence regional vascular adaptation. This thesis investigated the acute response and chronic adaptations of the peripheral vasculature to low load resistance exercise with BFR. The methodology utilised Doppler ultrasound, strain gauge plethysmography and muscle biopsy for insightful measures of the vasculature at different regions of the arterial tree. Short term (4-6 weeks) localised low load (30-40% 1RM) resistance exercise with BFR increased brachial (3.1%) and popliteal (3.3%) artery maximal diameter (in response to ischemic exercise), forearm (29%) and calf (24%) post-occlusive blood flow, and calf filtration capacity (14%). These findings indicate potential vascular remodelling at the conduit (chapters 3, 4) resistance (chapter 4) and capillary (chapter 4) level of the vascular tree. Regional, rather than systemic, factors are responsible for these adaptations as evidenced by an absent response in the contralateral control limb. Transient improvements in popliteal artery FMD% occurred at week 2 before increased maximal diameter at week 6, suggesting functional changes precede structural remodelling (chapter 4). Maximal brachial artery diameter and forearm post-occlusive blood flow returned to baseline values after a 2 week detraining period, signifying rapid structural normalisation after stimulus removal (chapter 3). Enhanced capillarity, despite low training loads, could be explained by augmentation of VEGF (~7 fold), PGC-1α (~6 fold) and eNOS (~5 fold) mRNA, and upregulation VEGFR-2 (~5 fold) and HIF-1α (~2.5 fold) mRNA with BFR (chapter 5). This indicates a targeted angiogenic response potentially mediated through enhanced metabolic, ischemic and shear stress stimuli. Large between subject variability in the level of BFR was observed during upper and lower limb cuff inflation protocols. Adipose tissue thickness and mean arterial pressure were the largest independent determinants of upper and lower limb BFR, respectively (Chapter 6). In conclusion, this thesis demonstrates that low load resistance exercise with BFR induces adaptation in the conduit, resistance and capillary vessels. The mediators of this response are likely to be the hemodynamic and chemical signals elicited by repeated bouts of BFR resistance exercise, although confirmation of these mechanisms is required. The functional significance of these adaptations is unknown and warrants further investigation.

ISWI is a member of a versatile family of ATP-dependent chromatin remodelling complexes involved not only in transcription regulation (initiation, elongation and termination), but also in other cellular functions like maintenance of higher order chromatin structure and DNA replication. TbISWI, a novel ATPase of the ISWI family in Trypanosoma brucei, is involved in the transcriptional repression of silent VSG expression sites (ESs) in both bloodstream form (BF) and procyclic form (PF) life cycle stages of the parasite. Using in silico analysis, I have found that TbISWI is well conserved across the eukaryotic lineage, including those members of the order Kinetoplastida that do not exhibit antigenic variation. Compared to the ISWIs of higher eukaryotes, TbISWI has greater representation of random coils within its structure, an indicator of more structural fluidity and flexibility of interaction with multiple protein partners. Using an eGFP reporter based assay, I have studied the role of TbISWI in transcriptional repression of silent areas of the T. brucei genome. TbISWI was found to be involved in preventing inappropriate transcription of the silent VSG repertoires. TbISWI was also found to downregulate transcription in RNA pol I, but not pol II, transcription units. These results argue for the presence of at least two functionally distinct TbISWI complexes in T. brucei. Using DNA staining and fluorescence in situ hybridisation (FISH), I have investigated the potential effect of TbISWI depletion on cell cycle progression and minichromosome segregation. I did not find any evidence for the role of TbISWI in the maintenance of centromeric heterochromatin in T. brucei.

ATP-dependent chromatin remodellers represent a class of proteins that restructure chromatin through the action of a conserved helicase-like ATPase domain. Remodellers typically have several accessory binding domains alongside the ATPase. These confer target specificity and most commonly recognise histone post-translational modifications. SMARCAD1 is a ubiquitous chromatin remodeller involved with DNA replication and re- pair. It binds directly to PCNA at the site of DNA replication and recruits co-repressor KAP1 in order to silence newly produced chromatin. In contrast to most other chromatin remodellers, SMARCAD1 does not contain several different types of accessory domains. Only two CUE do- mains have been identified in addition to the SMARCAD1 core ATPase domain. CUE domains are a type of helical ubiquitin-binding domain. This thesis presents the findings of an investigation into the structure and function of the SMARCAD1 double CUE domain. The solution NMR structure is presented with results from NMR binding experiments mapped onto the structure. Each CUE domain was found to be an independent helix bundle connected by a dynamic flexible linker. The N-terminal CUE domain, CUE-1, binds ubiquitin and has an adjacent SUMO (a ubiquitin-like protein) binding motif on a protruding extended helix. The C-terminal CUE domain, CUE-2, has a very similar structure to several published CUE domains but does not bind ubiquitin due to a charged substitution at a highly conserved CUE consensus position. The SMARCAD1 double CUE domain binds KAP1 from nuclear extract and is likely to mediate the interaction between SMARCAD1 and KAP1. SMARCAD1 double CUE domain is not involved with PCNA binding.

Exercise training promotes a beneficial endothelial cell (EC) phenotype and results in conduit vessel adaptation. The specific underlying mechanisms have been proposed (shear stress, circumferential stress, hypoxia, metabolic) but are yet to be fully elucidated. This thesis investigated the predominant stimuli responsible for conduit vessel adaptation with training. Further, it developed a method of in situ EC extraction to allow for determination of the cellular and molecular mechanisms underpinning these adaptations. The methodology utilised two-dimensional (2D) Doppler ultrasound, strain gauge plethysmography, immunocytochemistry and RT-qPCR to provide insight in to vascular characteristics, predominantly of the brachial artery and peripheral EC. Long-term repeated isometric forearm muscle contractions as performed by well-trained rock climbers promoted greater resting, peak (in response to 5 min ischaemia) and maximal (in response to ischaemic exercise) brachial artery diameters compared with controls. This structural response is dependent upon confounders associated with exercise additional to shear stress as evidenced by the lack of brachial artery remodelling in response to 8 weeks of ischaemic preconditioning (IPC). A transient increase in flow-mediated dilation (FMD)% was observed following 6 weeks exposure to IPC, which became significant when controlled for baseline artery diameter, despite an absence of augmentation following long-term (≥ 8 weeks) exposure to a shear stimulus. This is in line with the suggested timeline of conduit vessel adaptation to exercise training of a transient increase in function at 2-4 weeks. Underpinning molecular mechanisms responsible were not determined but may be further investigated given that the endovascular biopsy technique was developed and improved in this thesis. The endovascular biopsy successfully yields approximately 2100 ± 1700 EC per sample, providing sufficient material for determination of expression of both mRNA (RT-qPCR) and protein (immunocytochemistry). Specifically, type 2 diabetics (T2DM) with symptomatic cardiac abnormalities exhibited augmented eNOS mRNA and protein in brachial artery EC as compared with non-diabetic controls with symptomatic cardiac abnormalities. In conclusion, this thesis demonstrates that although shear stress promotes a transient trend for enhancement in function of the peripheral conduit arteries, additional factors are required for long-term structural adaptations. Further, the endovascular biopsy technique offers a novel method of extracting and analysing EC for genes and proteins of interest to vascular health. The use of this technique to decipher the underlying cellular and molecular mechanisms involved in vascular adaptations with exercise requires further investigation.

During tissues development multipotent progenitor cells establish tissue-specific gene expression programmes, leading to differentiation into specialized cell types. It has been previously shown that the transcription factor p63, a master regulator of skin development, controls the expression of adhesion molecules and essential cytoskeleton components. It has also been shown that p63 plays an important role in establishing distinct three-dimensional conformations in the Epidermal Differentiation Complex (EDC) locus (Fessing et al., 2011). Here we show that in p63-null mice about 32% of keratinocytes showed altered nuclear morphology. Alterations in the nuclear shape were accompanied by decreased expression of nuclear lamins (Lamin A/C and Lamin B1), proteins of the LINC complex (Sun-1, nesprin-2/3) and Plectin. Plectin links components of the nuclear envelope (nesprin-3) with cytoskeleton and ChIP-qPCR assay with adult epidermal keratinocytes showed p63 binding to the consensus binding sequences on Plectin 1c, Sun-1 and Nesprin-3 promoters. As a possible consequence of the altered expression of nuclear lamins and nuclear envelope-associated proteins, changes in heterochromatin distribution as well as decrease of the expression of several polycomb proteins (Ezh2, Ring1B, Cbx4) has been observed in p63-null keratinocytes. Moreover, recent data in our lab have showed that p63 directly regulates Cbx4, a component of the polycomb PRC1 complex. Here we show that mice lacking Cbx4 displayed a skin phenotype, which partially resembles the one observed in p63-null mice with reduced epidermal thickness and keratinocyte proliferation. All together these data demonstrate that p63-regulated gene expression program in epidermal keratinocytes includes not only genes encoding adhesion molecules, cytoskeleton proteins (cytokeratins) and chromatin remodelling factors (Satb1, Brg1), but also polycomb proteins and components of the nuclear envelope, suggesting the existence of a functional link between cytoskeleton, nuclear architecture and three dimensional nuclear organization. Other proteins important for proper epidermal development and stratification, are cytokeratins. Here, we show that keratin genes play an essential role in spatial organization of other lineage-specific genes in keratinocytes during epidermal development. In fact, ablation of keratin type II locus from chromosome 15 in epidermal keratinocytes led to changes in the genomic organization with increased distance between the Loricrin gene located on chromosome 3 as well as between Satb1 gene located on chromosome 17 and keratin type II locus, resulting in a more peripheral localization of these genes in the nucleus. As a possible consequence of their peripheral localization, reduced expression of Loricrin and Satb1 has also been observed in keratins type II-deficient mice. These findings together with recent circularized chromosome conformation capture (4C) data, strongly suggest that keratin 5, Loricrin and Satb1 are part of the same interactome, which is required for the proper expression of these genes and proper epidermal development and epidermal barrier formation. Taken together these data suggest that higher order chromatin remodelling and spatial organization of genes in the nucleus are important for the establishment of lineage-specific differentiation programs in epidermal progenitor cells. These data provide an important background for further analyses of nuclear architecture in the alterations of epidermal differentiation, seen in pathological conditions, such as psoriasis and epithelial skin cancers.

Transcription of the ribosomal genes accounts for the majority of transcription in the cell due to the constant high demand for ribosomes. The number of proteins synthesized correlates with an effective ribosomal biogenesis, which is regulated by cell growth and proliferation. In the work presented in this thesis, we have investigated the ribosomal RNA genes 45S and 5S rRNA, which are transcribed by RNA Pol I and RNA Pol III, respectively. The focus of this work is the chromatin remodelling complex B-WICH, which is composed of WSTF, the ATPase SNF2h and NM1. We have studied in particular its role in ribosomal gene transcription. We showed in Study I that B-WICH is required to set the stage at rRNA gene promoters by remodelling the chromatin into an open, transcriptionally active configuration. This results in the binding of histone acetyl transferases to the genes and subsequent histone acetylation, which is needed for ribosomal gene activation. Study II investigated the role of B-WICH in transcription mediated by RNA polymerase III. We showed that B-WICH is essential to create an accessible chromatin atmosphere at 5S rRNA genes, which is compatible with the results obtained in Study 1. In this case, however, B-WICH operates as a licensing factor for c-Myc and the Myc/Max/Mxd network. Study III confirmed the importance and the function of the B-WICH complex as an activator of ribosomal genes. We demonstrated that B-WICH is important for the remodelling of the rDNA chromatin into an active, competent state in response to extracellular stimuli, and that the association of the B-WICH complex to the rRNA gene promoter is regulated by proliferative and metabolic changes in cells. The work presented in this thesis has confirmed that the B-WICH complex is an important regulator and activator of Pol I and Pol III transcription. We conclude that B-WICH is essential for remodelling the rDNA chromatin into a transcriptionally active state, as required for efficient ribosomal gene transcription. / <p>At the time of the doctoral defense, the following papers were unpublished and had a status as follows: Paper 2: Manuscript. Paper 3: Manuscript.</p><p> </p>

A asma é uma doença crônica das vias aéreas, cuja fisiopatologia compreende inflamação, hiper-responsividade e remodelação. As características clínicas e a espirometria são usadas para avaliar o controle da doença. Alguns asmáticos não conseguem controlar tomando altas doses de medicações de controle (asma de difícil controle). A tomografia computadorizada quantitativa de tórax (TCQ) é pouco usada na asma, mas pode fornecer biomarcadores da doença. Objetivos: Comparar os achados de TCQ de indivíduos sem doenças respiratórias (grupo controle - GC), com asma controlada (ABC) e com asma de difícil controle (ADC), visando prioritariamente parâmetros substitutivos do remodelamento das vias aéreas. Investigar se há correlações entre os parâmetros QCT e espirometria. Materiais e Métodos: Recrutamos sujeitos com ADC e ABC em um hospital terciário. Usamos registros médicos e convite pessoal para obter dados de CG. Todos os indivíduos foram submetidos a tomografia computadorizada de alta resolução e espirometria. Um software (Yacta) foi usado para obter dados de TCQ. O teste t de Student, o one-way ANOVA e o teste exato de Fisher foram realizados para comparar os dados dos grupos. Os coeficientes de correlação de Pearson foram calculados para avaliar as correlações entre os parâmetros espirométricos e TCQ. Resultados: O GC foi composto por 21 sujeitos; ABC, de 28 e ADC, de 27. Estes últimos eram mais idosos que GC e ABC (50,85 (10,11); 41,27 (11,57); 42,04 (10,11) anos; p = 0,002). A espessura relativa das vias aéreas (ERP3-8) foi significativamente diferente entre GC e ADC (45,31% (3,71); 49,38% (3,38); p = 0,001) e entre GC e ABC (45,31%, 71); 48,25 (4,51); p = 0,001). A espessura normalizada das vias aéreas (Pi10) foi significativamente diferente entre ADC e ABC (0,51 (0,10); 0,44 (0,10); p = 0,0001) e entre ADC e CG (0,51 (0,10), 0,39 (0,08), p = 0,0001). A área da parede das vias aéreas (AP/ASC-mm2) da terceira geração brônquica foi significativamente diferente entre o ADC e o GC (37,14 (9,70); 30,18 (5,06); p = 0,005) e entre ABC e GC (35, 54 (6,37); 30,18 (5,06); p = 0,005). A área da luz da via aérea da terceira geração brônquica (LA/ASC-mm2) foi significativamente diferente entre ADC e ABC (28,81 (10,08); 35,88 (7,12); p = 0,002) e entre ADC e GC (28 81 (10,08); 36,14 (7,29); p = 0,002). A atenuação máxima média (MMA-Hounsfield Units) da terceira geração brônquica foi significativamente diferente entre ADC e GC (-128,68 (67,15); -232,01 (75,20); p = 0,0001) e entre ABC e CG (-147,81 (75,31); -232,01 (75,20); p = 0,0001). Encontramos correlações negativas entre o volume expiratório forçado no primeiro segundo (VEF1) e a ERP na terceira, quarta e quinta gerações brônquicas; entre o índice Tiffeneau e a ERP nessas gerações; entre fluxo expiratório forçado (FEF25-75%) e ERP nas mesmas gerações. Encontramos correlações negativas entre VEF1 e AMMna terceira e quarta gerações e entre o índice de Tiffeneau e AMM nessas gerações. Encontramos correlações negativas entre o FEF(25-75%) e o AMM nessas gerações. Conclusões: Mesmo os asmáticos com doença controlada apresentaram evidências radiológicas de comprometimento da parede das vias aéreas; as evidências foram maiores em asma de difícil controle. Características tomográficas podem ser devido a inflamação ou remodelação. Encontramos correlações entre os parâmetros espirométricos e TCQ. Nossos achados reforçam a utilidade potencial dos parâmetros do TCQ como biomarcadores do controle da asma e da resposta ao tratamento, em pesquisa e em bases clínicas / Asthma is a chronic airway disease whose pathophysiology comprises inflammation, hyperresponsivenes and remodelling. Clinical characteristics and spirometry are used to assess disease control. Some asthmatics do not achieve control taking high doses of controller therapy (difficult to control asthma). Quantitative lung computed tomography (QCT) is seldom used in asthma but may provide asthma biomarkers. Objectives: To compare QCT findings of subjects without respiratory diseases (control group - CG), with controlled asthma (CA) and with difficult to control asthma (DCA), aiming primarily at surrogate parameters of airway remodelling. To investigate whether there are correlations between QCT and spirometry parameters. Materials and Methods: We recruited subjects with DCA and CA from a tertiary hospital. We used medical records and personal invitation to obtain CG data. All subjects underwent high resolution computed tomography and spirometry. A software (Yacta) was used to obtain QCT data. Student\'s t-test, one-way ANOVA and Fisher\'s exact test were performed to compare data from the groups. Pearson correlation coefficients were calculated to assess correlations between spirometric and QTC parameters. Results: The CG was comprised of 21 subjects; CA, of 28 and DCA, of 27. The latter had older subjects than CG and CA (50,85 (10,11); 41,27 (11,57); 42,04 (10,11) years; p=0,002). Relative airway thickness (RT) was significantly different between CG and DCA (45,31% (3,71); 49,38% (3,38); p=0,001) and between CG and CA (45,31% (3,71); 48,25 (4,51); p=0,001). Normalised airway thickness (Pi10) were significantly different between DCA and CA (0,51 (0,10); 0,44 (0,10); p=0,0001) and between DCA and CG (0,51 (0,10); 0,39 (0,08); p=0,0001). Airway wall area (AWAmm2) of the third bronchial generation was significantly different between DCA and CG (37,14 (9,70); 30,18 (5,06); p=0,005) and between CA and CG (35,54 (6,37); 30,18 (5,06); p=0,005). Airway lumen area of the third bronchial generation (ALA-mm2) was significantly different between DCA and CA (28,81 (10,08); 35,88 (7,12); p=0,002) and between DCA and CG (28,81 (10,08); 36,14 (7,29); p=0,002). Mean maximum attenuation (MMA-Hounsfield Units) of the third bronchial generation was significantly different between DCA and CG (-128,68 (67,15); -232,01 (75,20); p=0,0001) and between CA and CG (-147,81 (75,31); -232,01 (75,20); p=0,0001). We found negative correlations between forced expiratory volume on the first second (FEV1) and RT on the third, fourth and fifth bronchial generations ; between the Tiffeneau index and RT on those generations; between forced expiratory flow (FEF25-75) and RT on the same generations. We found negative correlations between FEV1 and MMA on the third and fourth generations and between the Tiffeneau index and MMA on those geneations. We found negative correlations between FEF25-75 and MMA on those generations.Conclusions: Even asthmatics with controlled disease had radiological evidence of airway wall involvement; the evidences were greater in difficult to control asthma. Tomographic features can be due to inflammation or remodelling. We found correlations between spirometric and QTC parameters. Our findings reinforce the potential usefulness of QTC parameters as biomarkers of asthma control and of response to treatment, on research and clinical grounds