Stochastic model of transcription initiation of closely spaced promoters in escherichia coli

Dissertação para obtenção do Grau de Mestre em Engenharia Biomédica

Ankle anticipatory postural adjustments during gait initiation in healthy and post-stroke subjects

Background: Anticipatory postural adjustments during gait initiation have an important role in postural stability but also in gait performance. However, these first phase mechanisms of gait initiation have received little attention, particularly in subcortical post-stroke subjects, where bilateral postural control pathways can be impaired. This study aims to evaluate ankle anticipatory postural adjustments during gait initiation in chronic post-stroke subjects with lesion in the territory of middle cerebral artery. Methods: Eleven subjects with post-stroke hemiparesis with the ability to walk independently and twelve healthy controls participated in this study. Bilateral electromyographic activity of tibialis anterior, soleus and medial gastrocnemius was collected during gait initiation to assess the muscle onset timing, period of activation/deactivation and magnitude of muscle activity during postural phase of gait initiation. This phase was identified through centre of pressure signal. Findings: Post-stroke group presented only half of the tibialis anterior relative magnitude observed in healthy subjects in contralesional limb (t=2.38, p=0.027) and decreased soleus deactivation period (contralesional limb, t=2.25, p=0.04; ipsilesional limb, t=3.67, p=0.003) as well its onset timing (contralesional limb, t=3.2. p=0.005; ipsilesional limb, t=2.88, p=0.033) in both limbs. A decreased centre of pressure displacement backward (t=3.45, p=0.002) and toward the first swing limb (t=3.29, p=0.004) was observed in post-stroke subjects. Interpretation: These findings indicate that chronic post-stroke subjects with lesion at middle cerebral artery territory present dysfunction in ankle anticipatory postural adjustments in both limbs during gait initiation.

Reliability of Two Methods for Identifying the Postural Phase of Gait Initiation in Healthy and Post-stroke Subjects

This study aims to compare two methods of assessing the postural phase of gait initiation as to intrasession reliability, in healthy and post-stroke subjects. As a secondary aim, this study aims to analyse anticipatory postural adjustments during gait initiation based on the centre of pressure (CoP) displacements in post-stroke participants. The CoP signal was acquired during gait initiation in fifteen post-stroke subjects and twenty-three healthy controls. Postural phase was identified through a baseline-based method and a maximal displacement based method. In both healthy and post-stroke participants higher intra-class correlation coefficient and lower coefficient of variation values were obtained with the baseline-based method when compared to the maximal displacement based method. Post-stroke participants presented decreased CoP displacement backward and toward the first swing limb compared to controls when the baseline-based method was used. With the maximal displacement based method, there were differences between groups only regarding backward CoP displacement. Postural phase duration in medial-lateral direction was also increased in post-stroke participants when using the maximal displacement based method. The findings obtained indicate that the baseline-based method is more reliable detecting the onset of gait initiation in both groups, while the maximal displacement based method presents greater sensitivity for post-stroke participants.

Acute gouty arthritis as a manifestation of immune reconstitution inflammatory syndrome after initiation of antiretroviral therapy

Immune reconstitution inflammatory syndrome (IRIS) in HIV-infected subjects initiating antiretroviral therapy most commonly involves new or worsening manifestations of previously subclinical or overt infectious diseases. Reports of non-infectious IRIS are much less common but represent important diagnostic and treatment challenges. We report on a 34-year-old HIV-infected male patient with no history of gout who developed acute gouty arthritis in a single joint one month after initiating highly active antiretroviral therapy.

Self-initiation expatriation: A focus on Italian and Portuguese students

The purpose of this work project is to analyze the phenomenon of self-initiated expatriation (SIE) through its link to the Protean Career and Career Capital theories, focusing in particular on Italian and Portuguese students attending a Master in the business area. The main research questions are to understand the reasons driving the intention to expatriate, after the conclusion of the academic path, using three main categories (Adventure Motivation, Work Characteristic Motivation and instrumental Motivation) and the intention to repatriate. A sample of Italian and Portuguese students was obtained. Italians show a higher intention to expatriate relative to Portuguese; nevertheless, no other significant differences were found among the two populations, because of the similar cultural background and economic situation. Additionally, several heterogeneities were observed considering other clusters defined by Gender, Teaching Language of the Master and Past International Experiences, across the two nationalities. Furthermore, possible future researches and practical implications were discussed.

Slow clinical improvement after treatment initiation in Leishmania/HIV coinfected patients

INTRODUCTION: In Brazil there is a large area of overlap of visceral leishmaniasis (VL) and HIV infection, which favored a increased incidence of coinfection Leishmania/HIV. METHODS: This study evaluated 65 consecutive patients with VL and their clinical response to treatment in two health care settings in Belo Horizonte, Brazil. RESULTS: At baseline, the clinical picture was similar between both groups, although diarrhea and peripheral lymphadenomegaly were more frequent in HIV-infected subjects. HIV-positive patients had lower median blood lymphocyte counts (686/mm³ versus 948/mm³p = 0.004) and lower values of alanine aminotransferase (ALT) (48IU/L versus 75.6IU/L p = 0.016) than HIV-negative patients. HIV-positive status (hazard ratio = 0.423, p = 0.023) and anemia (HR = 0.205, p = 0.002) were independent negative predictors of complete clinical response following antileishmanial treatment initiation. CONCLUSIONS: This study reinforces that all patients with VL should be tested for HIV infection, regardless of their clinical picture. This practice would allow early recognition of coinfection with initiation of antiretroviral therapy and, possibly, reduction in treatment failure.

HOXA9 as a key regulator in glioma initiation, aggressiveness and response to therapy

Tese de Doutoramento em Ciências da Saúde

Predicting trabecular bone microdamage initiation and accumulation using a non-linear perfect damage model

Studies evaluating the mechanical behavior of the trabecular microstructure play an important role in our understanding of pathologies such as osteoporosis, and in increasing our understanding of bone fracture and bone adaptation. Understanding of such behavior in bone is important for predicting and providing early treatment of fractures. The objective of this study is to present a numerical model for studying the initiation and accumulation of trabecular bone microdamage in both the pre- and post-yield regions. A sub-region of human vertebral trabecular bone was analyzed using a uniformly loaded anatomically accurate microstructural three-dimensional finite element model. The evolution of trabecular bone microdamage was governed using a non-linear, modulus reduction, perfect damage approach derived from a generalized plasticity stress-strain law. The model introduced in this paper establishes a history of microdamage evolution in both the pre- and post-yield regions

Identification and characterization of novel molecular mechanisms involved in initiation and progression of myelination

Schwann cells synthesize a large amount of membrane that form a specialized structure called myelin that surrounds axons and facilitate the transmission of electrical signal along neurons in peripheral nervous system (PNS). Previous studies demonstrated that both Schwann cell differentiation and de-differentiation (in the situation of a nerve injury or demyelinating disease) are regulated by cell-intrinsic regulators including several transcription factors. In particular, the de-differentiation of mature Schwann cells is driven by the activation of multiple negative regulators of myelination including Sox2, c-Jun, Notch and Pax3, all usually expressed in immature Schwann cells and suppressed at the onset of myelination. In order to identify new regulators of myelination involved in the development of the PNS, we analyzed the gene-expression profiling data from developing PNS and from three models of demyelinating neuropathies. This analysis led to the identification of Sox4, a member of the Sox family of transcription factors, as a potential candidate. To characterize the molecular function of Sox4 in PNS, we generated two transgenic lines of mice, which overexpress Sox4 specifically in Schwann cells. Detailed analysis of these mice showed that the overexpression of Sox4 in Schwann cells causes a delay in progression of myelination between post-natal day 2 (P2) and P5. Our in vitro analysis suggested that Sox4 cDNA can be overexpressed while the protein translation is tightly regulated. Interestingly, we observed that Sox4 protein is stabilized in nerves of the CMT4C mouse, a model of the human neuropathy. We therefore crossed Sox4 transgenic mice with CMT4C mice and we observed that Sox4 overexpression exacerbated the neuropathy phenotype in these mice. While recognized as being crucial for the normal function of both neurons and myelinating glial cells, the processes that regulate the beginning of myelination and the nature of the neuro-glial cross-talk remains mostly unknown. In order to gain insight into the molecular pathways involved in the interactions between neurons and associated glial cells, we developed a neuron-glia co-culture system based on microfluidic chambers and successfully induced myelination in this system by ascorbic acid. Importantly, we observed that in addition to acting on Schwann cells, ascorbic acid also modulate neuronal/axonal NRG1/ErbB2-B3 signalling. The experimental setting used in our study thus allowed us to discover a novel phenomena of propagation for myelination in vitro. The further characterization of this event brought us to identify other compounds able to induce myelination: ADAMs secretases inhibitor GM6001 and cyclic-AMP. The results generated during my thesis project are therefore not only important for the advancement of our understanding of how the PNS works, but may also potentially help to develop new therapies aiming at improvement of PNS myelination under disease conditions. - Les cellules de Schwann synthétisent une grande quantité de membrane formant une structure spécialisée appelée myéline qui entoure les axones et facilite la transmission du signal électrique le long des neurones du système nerveux périphérique (SNP). Des études antérieures ont démontré que la différenciation et la dédifférenciation des cellules de Schwann (dans la situation d'une lésion nerveuse ou d'une maladie démyélinisante) sont régulées par des régulateurs cellulaires intrinsèques, incluant plusieurs facteurs de transcription. En particulier, la dédifférenciation des cellules de Schwann matures est contrôlée par l'activation de plusieurs régulateurs négatifs de la myélinisation dont Sox2, c-Jun, Notch et Pax3, tous habituellement exprimés dans des cellules de Schwann immatures et supprimés au début de la myélinisation. Afin d'identifier de nouveaux régulateurs de myélinisation impliqués dans le développement du SNP, nous avons analysé le profil d'expression génique durant le développement du SNP ainsi que dans trois modèles de neuropathies démyélinisantes. Cette analyse a mené à l'identification de Sox4, un membre de la famille des facteurs de transcription Sox, comme étant un candidat potentiel. Dans le but de caractériser la fonction moléculaire de Sox4 dans le SNP, nous avons généré deux lignées transgéniques de souris qui surexpriment Sox4 spécifiquement dans les cellules de Schwann. L'analyse détaillée de ces souris a montré que la surexpression de Sox4 dans les cellules de Schwann provoque un retard dans la progression de la myélinisation entre le jour postnatal 2 (P2) et P5. Notre analyse in vitro a suggéré que l'ADNc de Sox4 peut être surexprimé alors que la traduction des protéines est quand à elle étroitement régulée. De façon intéressante, nous avons observé que la protéine Sox4 est stabilisée dans les nerfs des souris CMT4C, un modèle de neuropathie humaine. Nous avons donc croisé les souris transgéniques Sox4 avec des souris CMT4C et avons observé que la surexpression de Sox4 exacerbe le phénotype de neuropathie chez ces souris. Bien que reconnus comme étant cruciaux pour le fonctionnement normal des neurones et des cellules gliales myélinisantes, les processus qui régulent le début de la myélinisation ainsi que la nature des interactions neurone-glie restent largement méconnus. Afin de mieux comprendre les mécanismes moléculaires impliqués dans les interactions entre les neurones et les cellules gliales leur étant associés, nous avons développé un système de co-culture neurone-glie basé sur des chambres microfluidiques et y avons induit avec succès la myélinisation avec de l'acide ascorbique. Étonnamment, nous avons remarqué que, en plus d'agir sur les cellules de Schwann, l'acide ascorbique module également la voie de signalisation neuronale/axonale NRG1/ErbB2-B3. Le protocole expérimental utilisé dans notre étude a ainsi permis de découvrir un nouveau phénomène de propagation de la myélinisation in vitro. La caractérisation plus poussée de ce phénomène nous a menés à identifier d'autres composés capables d'induire la myélinisation: L'inhibiteur de sécrétases ADAMs GM6001 et l'AMP cyclique. Les résultats obtenus au cours de mon projet de thèse ne sont donc pas seulement importants pour l'avancement de notre compréhension sur la façon dont le SNP fonctionne, mais peuvent aussi potentiellement aider à développer de nouvelles thérapies visant à l'amélioration de la myélinisation du SNP dans des conditions pathologiques.

Canakinumab Reduces The Risk Of Acute Gouty Arthritis Flares During Initiation Of Allopurinol Therapy: Results Of A Double-blind, Randomised Study

RATIONALE: This study assessed the efficacy and safety of canakinumab, a fully human anti-interleukin-1beta monoclonal antibody, for prophylaxis against acute gouty arthritis flares in patients initiating uratelowering therapy.METHODS: In this double-blind, double-dummy, dose-ranging study, 432 patients with gouty arthritis initiating allopurinol therapy were randomised 1:1:1:1:1:1:2 to receive: a single dose of canakinumab, 25, 50, 100, 200, or 300 mg subcutaneously (sc); four 4-weekly doses of canakinumab (50150125125 mg sc); or daily colchicine 0.5 mg orally for 16 weeks. Patients recorded details of flares in diaries. The study aimed to determine the canakinumab dose having equivalent efficacy to colchicine 0.5 mg at 16 weeks.RESULTS: A dose-response for canakinumab was not apparent with any of the four pre-defined dose-responsemodels. The estimated canakinumab dose with equivalent efficacy to colchicinewas belowthe range of doses tested.At 16 weeks, therewas a 62-72% reduction in themean number of flares per patient for canakinumab doses >50 mg vs colchicine based on a negative binomial model (rate ratio: 0.28-0.38, p50.0083), and the percentage of patients experiencing >1 flarewas significantly lower for all canakinumab doses (15- 27%) vs colchicine (44%, p<0.05). Therewas a 64-72%reduction in the risk of experiencing >1 flare for canakinumab doses >50 mg vs colchicine at 16 weeks (hazard ratio: 0.28-0.36, p50.05). The incidence of adverse events was similar across treatment groups.CONCLUSIONS: Single canakinumab doses >50 mg or four 4-weekly doses provided superior prophylaxis against flares compared with daily colchicine 0.5 mg.

Defining the site of light perception and initiation of phototropism in Arabidopsis.

Phototropism is an adaptive response allowing plants to optimize photosynthetic light capture. This is achieved by asymmetric growth between the shaded and lit sides of the stimulated organ. In grass seedlings, the site of phototropin-mediated light perception is distinct from the site of bending; however, in dicotyledonous plants (e.g., Arabidopsis), spatial aspects of perception remain debatable. We use morphological studies and genetics to show that phototropism can occur in the absence of the root, lower hypocotyl, hypocotyl apex, and cotyledons. Tissue-specific expression of the phototropin1 (phot1) photoreceptor demonstrates that light sensing occurs in the upper hypocotyl and that expression of phot1 in the hypocotyl elongation zone is sufficient to enable a normal phototropic response. Moreover, we show that efficient phototropism occurs when phot1 is expressed from endodermal, cortical, or epidermal cells and that its local activation rapidly leads to a global response throughout the seedling. We propose that spatial aspects in the steps leading from light perception to growth reorientation during phototropism differ between grasses and dicots. These results are important to properly interpret genetic experiments and establish a model connecting light perception to the growth response, including cellular and morphological aspects.

Subduction and obduction processes in the Swiss Alps

The significance of the Brianconnais domain in the Alpine orogen is reviewed in the light of data concerning its collision with the active Adriatic margin and the passive Helvetic margin. The Brianconnais which formerly belonged to the Iberian plate, was located on the northern margin of the Alpine Tethys (Liguro-Piemont ocean) since its opening in the early-Middle Jurassic. Together with the Iberian plate the Brianconnais terrane was separated from the European plate in the Late Jurassic-Early Cretaceous, following the northern Atlantic, Bay of Biscay, Valais ocean opening. This was accompanied by the onset of subduction along the northern margin of Adria and the closure of the Alpine Tethys. Stratigraphic and metamorphic data regarding this subduction and the geohistory of the Brianconnais allows the scenario of subduction-obduction processes during the Late Cretaceous-early Tertiary in the eastern and western Alps to be specified. HP-LT metamorphism record a long-lasting history of oceanic subduction-accretion, followed in the Middle Eocene by the incorporation of the Brianconnais as an exotic terrane into the accretionary prism. Middle to Late Eocene cooling ages of the Brianconnais basement and the presence of pelagic, anorogenic sedimentation lasting until the Middle Eocene on the Brianconnais preclude any sort of collision before that time between this domain and the active Adria margin or the Helvetic margin. This is confirmed by plate reconstructions constrained by magnetic anomalies in the Atlantic domain. Only a small percentage of the former Brianconnais domain was obducted, most of the crust and lithospheric roots were subducted. This applies also to domains formerly belonging to the southern Alpine Tethys margin (Austroalpine-inner Carpathian domain). It is proposed that there was a single Palaeogene subduction zone responsible for the Alpine orogen formation (from northern Spain to the East Carpathians), with the exception of a short-lived Late Cretaceous partial closure of the Valais ocean. Subduction in the western Tethyan domain originated during the closure of the Meliata ocean during the Jurassic incorporating the Austroalpine-Carpathian domain as terranes during the Cretaceous. The subduction zone propagated into the northern margin of Adria and then to the northern margin of the Iberian plate, where it gave birth to the Pyrenean-Provencal orogenic belt. This implies the absence of a separated Cretaceous subduction zone within the Austro-Carpathian Penninic ocean. Collision of Iberia with Europe forced the subduction to jump to the SE margin of Iberia in the Eocene, creating the Apenninic orogenic wedge and inverting the vergence of subduction from south- to north-directed. (C) 1998 Elsevier Science B.V. All rights reserved.

Initiation of primary cell cultures from embryos of the mosquitoes Anopheles albimanus and Aedes taeniorhynchus (Diptera: Culicidae)

Primary cell cultures were obtained from eggs of Anopheles albimanus and Aedes taeniorhynchus mosquitoes, vectors of human malaria and of Venezuelan equine encephalitis virus, respectively. The cellular growth of the An. albimanus cells began four weeks after explanting the embryonic tissues in MK/VP12 medium, supplemented with 15% fetal bovine serum. The culture showed heterogeneous cellular morphology. With regard to the Ae. taeniorhynchus culture, growth occurred three weeks after initiating the culture in MM/VP12 medium. The majority of cells were small and round. Karyotypes were examined in the latter species.

La Bible se raconte : initiation à l'analyse narrative

La Bible est l'un des plus fabuleux trésors d'histoires de l'humanité. Car depuis la nuit des temps, les croyants racontent. Mais pas n'importe quoi. Longuement façonnés dans l'oralité, patiemment rédigés, les récits bibliques obéissent à de subtiles règles de composition. Derrière les histoires apparemment naives se cache la fine stratégie d'un narrateur. Grâce à ses qualités pédagogiques, le livre de Daniel Marguerat et Yvan Bourquin s'est imposé comme un manuel de référence pour l'initiation à l'analyse narrative des textes bibliques. Il a été traduit en plusieurs langue. Pour cette 4e édition, l'étude de l'intrigue et de la construction des personnages a été renouvelée. Avec clarté, ses auteurs ont aménagé un parcours qui permet de découvrir les outils de l'analyse narrative, d'étudier leur application, d'évaluer leur efficacité. Toute la magie de la lecture biblique s'en trouve éclairée.