Regulation of MAPK activation, AP-1 transcription factor expression and keratinocyte differentiation in wounded fetal skin

Fetal epithelium retains the ability to re-epithelialize a wound in organotypic culture in a manner not dependent on the presence of underlying dermal substrata. This capacity is lost late in the third trimester of gestation or after embryonic day 17 (E-17) in the rat such that embryonic day 19 (E-19) wounds do not re-epithelialize. Moreover, wounds created in E-17 fetuses in utero heal in a regenerative, scar-free fashion. To investigate the molecular events regulating re-epithelialization in fetal skin, the wound-induced expression profile and tissue localization of activator protein 1 (AP-1) transcription factors c-Fos and c-Jun was characterised in E-17 and E-19 skin using organotypic fetal cultures. The involvement of mitogen-activated protein kinase (MAPK) signaling in mediating wound-induced transcription factor expression and wound re-epithelialization was assessed, with the effect of wounding on the expression of keratinocyte differentiation markers determined. Our results show that expression of AP-1 transcription factors was induced immediately by wounding and localized predominantly to the epidermis in E-17 and E-19 skin. c-fos and c-jun induction was transient in E-17 skin with MAPK-dependent c-fos expression necessary for the re-epithelialization of an excisional wound in organotypic culture. In E-19 skin, AP-11 expression persisted beyond 12 h post-wounding, and marked upregulation of the keratinocyte differentiation markers keratin 10 and loricrin was observed. No such changes in the expression of keratin 10 or loricrin occurred in E-17 skin. These findings indicate that re-epithelialization in fetal skin is regulated by wound-induced AP-1 transcription factor expression via MAPK and the differentiation status of keratinocytes.

CULTO IMPERIAL E O APOCALIPSE DE JOÃO Uma análise exegética de Ap 13,1-18

Esta dissertação investiga a narrativa de visão apocalíptica encontrada em Ap 13,1-18. Ela parte da pergunta sobre a realidade que influenciou o autor no momento da composição, utilizando-se, assim, de uma linguagem provocativa. Nossa hipótese indica que o Apocalipse de João, importante fonte das experiências do Cristianismo primitivo no final do primeiro século, proporciona uma dura crítica às exigências de adoração dirigidas às autoridades romanas através do Culto Imperial. As imagens das bestas descritas em Ap 13,1-18 expressam este tema através de uma linguagem provocativa, fundamentada na força da palavra e na tradição do mito do antagonista encontrado no Antigo Oriente Próximo. Neste sentido, acreditamos que o autor demoniza e estigmatiza as expressões da religião oficial, as quais tratam os governantes como seres divinos, e os promotores do Culto Imperial na região da Ásia Menor.(AU)

O TRATAMENTO CONTABILÍSTICO DOS ATIVOS INTANGÍVEIS: SNC versus SNC-AP

O objetivo central deste trabalho é efetuar uma análise comparativa da contabilização dos ativos intangíveis no âmbito do SNC e do SNC-AP. A convergência de ambos os normativos nos critérios de reconhecimento e mensuração não é plena. No que respeita ao reconhecimento, é dada a possibilidade, no âmbito público, de um elemento ser reconhecido como ativo, mesmo que não produza benefícios económicos futuros para a entidade, desde que possua potencial de serviço. No que à mensuração se refere, na norma aplicada ao setor público o modelo da revalorização é visto como uma alternativa ao modelo do custo, não estando previsto nesse modelo o reconhecimento de perdas por imparidade. As conclusões observadas neste trabalho poderão contribuir para que outros estudos se desenvolvam no âmbito de uma análise conjunta aos normativos contabilísticos adotados em Portugal.

ESTUDO COMPARATIVO DO RECONHECIMENTO E DA MENSURAÇÃO DOS ATIVOS FIXOS TANGÍVEIS NO POCAL E NO SNC-AP

Perante a atual reforma da Contabilidade Pública em Portugal, foi aprovado, em setembro de 2015, o novo Sistema de Normalização Contabilística para as Administrações Públicas (SNC-AP). Este trabalho tem por objetivos analisar o definido no POCAL e no SNC-AP quanto ao reconhecimento e à mensuração dos ativos fixos tangíveis (AFT). Este estudo permite concluir que o novo normativo evidencia um avanço face ao POCAL, definindo não só o conceito de ativo e de AFT, como também os critérios de reconhecimento que um elemento deve cumprir para que possa ser reconhecido como tal, permitindo assim uma maior consistência no reconhecimento destes elementos, por parte das diferentes entidades públicas. Por outro lado, no que respeita à mensuração, o SNC-AP segue de perto, com as devidas adaptações, os normativos internacionais, introduzindo dois momentos de mensuração dos AFT, e referindo o justo valor explicitamente enquanto critério de mensuração aplicável a situações concretas.

Ascorbic acid induces an oxidative stress in CCRF cells activating the transcription factors AP-1 and NFRB

We have previously tested the effects of high dose AA supplements on human volunteers in terms of reducing DNA damage, as a possible mechanism of the vitamin’s proposed protective effect against cancer and detected a transient, pro-oxidant effect at high doses (500 mg/day). Herein, we present evidence of a pro-oxidant effect of the vitamin when added to CCRF cells at extracellular concentrations which mimic those present in human serum in vivo (50–150AM). The activation of the transcription factor AP-1 was optimal at 100 AM AA following 3h exposure at 37jC. A minimum dose of 50 AM of AA activated NFnB but there appeared to be no dose-dependent effect. Increases of 2–3 fold were observed for both transcription factors when cells were exposed to 100 AM AA for 3h, comparing well with the pro-oxidant effect of H2O2 at similar concentrations. In parallel experiments the activation of AP-1 (binding to DNA) was potentiated when cells were pre-incubated with AA prior to exposure with H2O2. Cycloheximide pretreatment (10 Ag/ml for 15min) caused a 50% inhibition of AP-1 binding to DNA suggesting that it was due to a combination of increasing the binding of pre-existing Fos and Jun and an increase in their de novo synthesis. Cellular localisation was confirmed by immunocytochemistry using antibodies specific for c-Fos and c-Jun proteins. These results suggest that extracellular AA can elicit an intracellular stress response resulting in the activation of the oxidative stress-responsive transcription factors AP-1 and NFnB. These transcription factors are involved in the induction of genes associated with an oxidative stress response, cell cycle arrest and DNA repair confirmed by our cDNA microarray analysis (Affymetrix). This may explain the abilty for AA to appear to inhibit 8-oxodG, yet simultaneously generate another oxidative stress biomarker, 8-oxo-dA. These results suggest a completely novel DNA repair action for AA. Whether this action is relevant to our in vivo findings will be the subject of our future research.

Septin6 and Septin7 GTP binding proteins regulate AP-3- and ESCRT-dependent multivesicular body biogenesis

Septins (SEPTs) form a family of GTP-binding proteins implicated in cytoskeleton and membrane organization, cell division and host/pathogen interactions. The precise function of many family members remains elusive. We show that SEPT6 and SEPT7 complexes bound to F-actin regulate protein sorting during multivesicular body (MVB) biogenesis. These complexes bind AP-3, an adapter complex sorting cargos destined to remain in outer membranes of maturing endosomes, modulate AP-3 membrane interactions and the motility of AP-3-positive endosomes. These SEPT-AP interactions also influence the membrane interaction of ESCRT (endosomal-sorting complex required for transport)-I, which selects ubiquitinated cargos for degradation inside MVBs. Whereas our findings demonstrate that SEPT6 and SEPT7 function in the spatial, temporal organization of AP-3- and ESCRT-coated membrane domains, they uncover an unsuspected coordination of these sorting machineries during MVB biogenesis. This requires the E3 ubiquitin ligase LRSAM1, an AP-3 interactor regulating ESCRT-I sorting activity and whose mutations are linked with Charcot-Marie-Tooth neuropathies.

AP-2 complex-mediated endocytosis of Drosophila Crumbs regulates polarity via antagonizing Stardust

Maintenance of epithelial polarity depends on the correct localization and levels of polarity determinants. The evolutionarily conserved transmembrane protein Crumbs is crucial for the size and identity of the apical membrane, yet little is known about the molecular mechanisms controlling the amount of Crumbs at the surface. Here, we show that Crumbs levels on the apical membrane depend on a well-balanced state of endocytosis and stabilization. The adaptor protein 2 (AP-2) complex binds to a motif in the cytoplasmic tail of Crumbs that overlaps with the binding site of Stardust, a protein known to stabilize Crumbs on the surface. Preventing endocytosis by mutations in AP-2 causes expansion of the Crumbs-positive plasma membrane and polarity defects, which can be partially rescued by removing one copy of crumbs. Strikingly, knocking-down both AP-2 and Stardust retains Crumbs on the membrane. This study provides evidence for a molecular mechanism, based on stabilization and endocytosis, to adjust surface levels of Crumbs, which are essential for maintaining epithelial polarity.

An Examination of the AP United States History Exam Free Response Section

Michael S. Henry examined the first 30 years of the AP United States History exam’s essay section. This study examined changes that have occurred over the last 20 years by classifying questions into one of six categories and found little change in the types of essays used during this timeframe.

Classificador máquina de suporte vetorial com análise de Fourier aplicada em dados de EEG e EMG

The classifier support vector machine is used in several problems in various areas of knowledge. Basically the method used in this classier is to end the hyperplane that maximizes the distance between the groups, to increase the generalization of the classifier. In this work, we treated some problems of binary classification of data obtained by electroencephalography (EEG) and electromyography (EMG) using Support Vector Machine with some complementary techniques, such as: Principal Component Analysis to identify the active regions of the brain, the periodogram method which is obtained by Fourier analysis to help discriminate between groups and Simple Moving Average to eliminate some of the existing noise in the data. It was developed two functions in the software R, for the realization of training tasks and classification. Also, it was proposed two weights systems and a summarized measure to help on deciding in classification of groups. The application of these techniques, weights and the summarized measure in the classier, showed quite satisfactory results, where the best results were an average rate of 95.31% to visual stimuli data, 100% of correct classification for epilepsy data and rates of 91.22% and 96.89% to object motion data for two subjects.

Construção de um classificador automático de severidade de bugs para sistemas open source

Software bug analysis is one of the most important activities in Software Quality. The rapid and correct implementation of the necessary repair influence both developers, who must leave the fully functioning software, and users, who need to perform their daily tasks. In this context, if there is an incorrect classification of bugs, there may be unwanted situations. One of the main factors to be assigned bugs in the act of its initial report is severity, which lives up to the urgency of correcting that problem. In this scenario, we identified in datasets with data extracted from five open source systems (Apache, Eclipse, Kernel, Mozilla and Open Office), that there is an irregular distribution of bugs with respect to existing severities, which is an early sign of misclassification. In the dataset analyzed, exists a rate of about 85% bugs being ranked with normal severity. Therefore, this classification rate can have a negative influence on software development context, where the misclassified bug can be allocated to a developer with little experience to solve it and thus the correction of the same may take longer, or even generate a incorrect implementation. Several studies in the literature have disregarded the normal bugs, working only with the portion of bugs considered severe or not severe initially. This work aimed to investigate this portion of the data, with the purpose of identifying whether the normal severity reflects the real impact and urgency, to investigate if there are bugs (initially classified as normal) that could be classified with other severity, and to assess if there are impacts for developers in this context. For this, an automatic classifier was developed, which was based on three algorithms (Näive Bayes, Max Ent and Winnow) to assess if normal severity is correct for the bugs categorized initially with this severity. The algorithms presented accuracy of about 80%, and showed that between 21% and 36% of the bugs should have been classified differently (depending on the algorithm), which represents somewhere between 70,000 and 130,000 bugs of the dataset.