Interação da cianotoxina microcistina e o nanomaterial de carbono fulereno (C60) em brânquias do peixe Cyprinus carpio (Teleostei: Cyprinidae) sob incidência de radiação UVA

A produção mundial de nanomateriais tem aumentado nos últimos anos, em função de suas variadas aplicações tecnológicas e, como consequência do seu crescente uso e demanda, poderão existir riscos ambientais sendo a água o ambiente onde muitas destas substâncias podem exercer efeitos deletérios. Um dos nanomaterias de carbono mais utilizados é o fulereno, um composto orgânico lipofílico que pode se comportar como carreador de moléculas tóxicas, potencializando a entrada de contaminantes ambientais em órgãos específicos, fenômeno conhecido como “cavalo de Troia”. As microcistinas (MC) são cianotoxinas produzidas por cianobactérias durante episódios de floração, afetando aos organismos aquáticos e ao ser humano. Diversos estudos demonstram que organismos expostos tanto às MCs quanto ao fulereno podem causar produção excessiva de espécies ativas de oxigênio e alterar os níveis de antioxidantes. Além disso, outro fator que pode vir a intensificar o potencial tóxico de ambos é a incidência de radiação UVA. Sendo assim, procurou-se avaliar os efeitos em parâmetros de estresse oxidativo da co-exposição ex vivo da cianotoxina microcistina-LR (MC-LR) e o nanomaterial de carbono fulereno em brânquias do peixe Cyprinus carpio sob incidência de radiação UVA. Os resultados mostraram que: (a) houve uma perda da capacidade antioxidante no tratamento com MC-LR (baixa concentração) quando coexposta com fulereno no UVA em relação com o tratamento realizado sem co-exposição com fulereno; (b) o fulereno no UV diminuiu a atividade da enzima glutationa-Stransferase (GST) quando comparado com o controle no UV; (c) a MC-LR (alta concentração) co-exposta com fulereno foi capaz de diminuir as concentrações do antioxidante glutationa (GSH) quando comparado com o mesmo tratamento tanto no UVA quanto no escuro sem a co-exposição ao fulereno; (d) o tratamento MC-LR (baixa concentração) com UVA aumentou o dano oxidativo lipídico quando comparado com o controle UVA; (e) o fulereno não causou uma maior bioacumulação da microcistina no tecido. Sendo assim, pode-se concluir que o fulereno não apresentou o potencial de carregador de moléculas nessas concentrações de microcistina, porém, a co-exposição dos compostos diminuem tanto capacidade antioxidante total, como a concentração da GSH, podendo gerar problemas a longo prazo na detoxificação da toxina.

Outcome Prediction of Consciousness Disorders in the Acute Stage Based on a Complementary Motor Behavioural Tool.

INTRODUCTION: Attaining an accurate diagnosis in the acute phase for severely brain-damaged patients presenting Disorders of Consciousness (DOC) is crucial for prognostic validity; such a diagnosis determines further medical management, in terms of therapeutic choices and end-of-life decisions. However, DOC evaluation based on validated scales, such as the Revised Coma Recovery Scale (CRS-R), can lead to an underestimation of consciousness and to frequent misdiagnoses particularly in cases of cognitive motor dissociation due to other aetiologies. The purpose of this study is to determine the clinical signs that lead to a more accurate consciousness assessment allowing more reliable outcome prediction. METHODS: From the Unit of Acute Neurorehabilitation (University Hospital, Lausanne, Switzerland) between 2011 and 2014, we enrolled 33 DOC patients with a DOC diagnosis according to the CRS-R that had been established within 28 days of brain damage. The first CRS-R assessment established the initial diagnosis of Unresponsive Wakefulness Syndrome (UWS) in 20 patients and a Minimally Consciousness State (MCS) in the remaining13 patients. We clinically evaluated the patients over time using the CRS-R scale and concurrently from the beginning with complementary clinical items of a new observational Motor Behaviour Tool (MBT). Primary endpoint was outcome at unit discharge distinguishing two main classes of patients (DOC patients having emerged from DOC and those remaining in DOC) and 6 subclasses detailing the outcome of UWS and MCS patients, respectively. Based on CRS-R and MBT scores assessed separately and jointly, statistical testing was performed in the acute phase using a non-parametric Mann-Whitney U test; longitudinal CRS-R data were modelled with a Generalized Linear Model. RESULTS: Fifty-five per cent of the UWS patients and 77% of the MCS patients had emerged from DOC. First, statistical prediction of the first CRS-R scores did not permit outcome differentiation between classes; longitudinal regression modelling of the CRS-R data identified distinct outcome evolution, but not earlier than 19 days. Second, the MBT yielded a significant outcome predictability in the acute phase (p<0.02, sensitivity>0.81). Third, a statistical comparison of the CRS-R subscales weighted by MBT became significantly predictive for DOC outcome (p<0.02). DISCUSSION: The association of MBT and CRS-R scoring improves significantly the evaluation of consciousness and the predictability of outcome in the acute phase. Subtle motor behaviour assessment provides accurate insight into the amount and the content of consciousness even in the case of cognitive motor dissociation.

The TOMO-ETNA experiment: an imaging active campaign at Mt. Etna volcano. Context, main objectives, working-plans and involved research projects

The TOMO-ETNA experiment was devised to image of the crust underlying the volcanic edifice and, possibly, its plumbing system by using passive and active refraction/reflection seismic methods. This experiment included activities both on-land and offshore with the main objective of obtaining a new high-resolution seismic tomography to improve the knowledge of the crustal structures existing beneath the Etna volcano and northeast Sicily up to Aeolian Islands. The TOMO ETNA experiment was divided in two phases. The first phase started on June 15, 2014 and finalized on July 24, 2014, with the withdrawal of two removable seismic networks (a Short Period Network and a Broadband network composed by 80 and 20 stations respectively) deployed at Etna volcano and surrounding areas. During this first phase the oceanographic research vessel “Sarmiento de Gamboa” and the hydro-oceanographic vessel “Galatea” performed the offshore activities, which includes the deployment of ocean bottom seismometers (OBS), air-gun shooting for Wide Angle Seismic refraction (WAS), Multi-Channel Seismic (MCS) reflection surveys, magnetic surveys and ROV (Remotely Operated Vehicle) dives. This phase finished with the recovery of the short period seismic network. In the second phase the Broadband seismic network remained operative until October 28, 2014, and the R/V “Aegaeo” performed additional MCS surveys during November 19-27, 2014. Overall, the information deriving from TOMO-ETNA experiment could provide the answer to many uncertainties that have arisen while exploiting the large amount of data provided by the cutting-edge monitoring systems of Etna volcano and seismogenic area of eastern Sicily.

A monitorização da carga de treino através da análise tempo-movimento e da perceção subjetiva do esforço, PSE, no futebol

O objetivo do presente estudo foi verificar se existe variação dos parâmetros que caraterizam a atividade motora de jogadores de futebol em treino, obtidos através do Viper Software STATSports, ao longo de vários microciclos (MCs) integrando o período preparatório (PP) e o início do período competitivo (PC), assim como, averiguar a associação existente entre a variação destes parâmetros, MC a MC, e a Perceção Subjetiva do Esforço Sessão (PSE-sessão) respetiva, e verificar, ainda, se existe correlação entre estes parâmetros e a PSE, em cada MC do período considerado. A amostra do estudo foi constituída por 13 jogadores (idade: 25 ± 1,78 anos) de futebol do sexo masculino a competir no escalão de Séniores da Associação Futebol de Setúbal. A recolha de dados ocorreu durante o PP (4 semanas) e o PC (4 semanas). Os jogadores incluídos no estudo cumpriram o único critério definido: a realização total das sessões de treino previstas. Os instrumentos utilizados durante o processo foram o sistema Viper (Viper Software v.1.2, STATSports), integrando um GPS 10 Hz, acelerómetro 100 Hz, cardiofrequencímetro e a escala de PSE adaptada por Foster. Verificou-se elevada variabilidade no comportamento dos parâmetros no PP, mas não nos 3 últimos MCs já integrados no PC. Verificamos forte associação entre os parâmetros Distância Explosiva (r=0,762), Nº Esforços de Elevada Carga Metabólica (r=0,714), Nº Acelerações (r=0,714), Nº Desacelerações (r=0,762) e a PSE-sessão (p˂0,05). O parâmetro com maior potencial preditor de elevados valores da PSE-sessão é a Distância Explosiva (p˂0,001). Por fim, não encontramos uma associação regular entre os parâmetros e a PSE-sessão em cada MC. Os valores de PSE-sessão apresentados durante o estudo são semelhantes a valores apresentados em outros estudos no futebol profissional. Conclui-se que no PP ocorreu a variabilidade de todos os parâmetros e estabilização no PC da Distância Total, Distância de Elevada Carga Metabólica, Distâcia Explosiva, Nº Esforços de Elevada Carga Metabólica e PSE-sessão. A PSE-sessão depende dos parâmetros Distância Explosiva, Nº Esforços de Elevada Carga Metabólica, Nº Acelerações e Nº Desacelerações, i.e., as ações de elevada exigência metabólica e neuromuscular explicam a relação existente entre os parâmetros de carga e a PSE-sessão como indicador global de carga interna. A utilização do sistema Viper possibilita a recolha de parâmetros que descrevem as exigências fisícas a que os jogadores estão sujeitos e fornece informação fundamental para o planeamento do treino em conjunto com a recolha sistemática da PSE-sessão.

Produção de pellets de madeira no Brasil : estratégia, custo e risco do investimento

Tese (doutorado)—Universidade de Brasília, Faculdade de Tecnologia, Departamento de Engenharia Florestal, Programa de Pós-Graduação em Ciências Florestais, 2015.

A implementação de preços de transferência interna : o caso Xerox

Mestrado em Contabilidade, Fiscalidade e Finanças Empresariais

Improving safety and establishing episomal maintenance of Adeno-associated viral vectors

Adeno-associated viral (AAV) vectors are among the most widely used gene transfer systems in basic and pre-clinical research and have been employed in more than 160 clinical trials. AAV vectors are commonly produced in producer cell lines like HEK293 by co-transfection with a so-called vector plasmid and one (in this work) or two so-called helper plasmids. The vector plasmid contains the transgene cassette of interest (TEC) flanked by AAV’s inverted terminal repeats (ITRs) which serve as packaging signals, whereas the helper plasmid provides the required AAV and helper virus functions in trans. A pivotal aspect of AAV vectorology is the manufacturing of AAV vectors free from impurities arising during the production process. These impurities include AAV vector preparations that contain capsids containing prokaryotic sequences, e.g. antibiotic resistance genes originating from the producer plasmids. In the first part of the thesis we aimed at improving the safety of AAV vectors. As we found that encapsidated prokaryotic sequences (using the ampicillin resistance gene as indicator) cannot be re-moved by standard purification methods we investigated whether the producer plasmids could be replaced by Minicircles (MCs). MCs are circular DNA constructs which contain no functional or coding prokaryotic sequences; they only consist of the TEC and a short sequence required for production and purification. MC counterparts of a vector plasmid encoding for enhanced green fluorescent (eGFP) protein and a helper plasmid encoding for AAV serotype 2 (AAV2) and helper Adenovirus (Ad) genes were designed and produced by PlasmidFactory (Bielefeld, Germany). Using all four possible combinations of plasmid and MCs, single-stranded AAV2 vectors (ssAAV) and self-complementary AAV vectors (scAAV) were produced and characterized for vector quantity, quality and functionality. The analyses showed that plasmids can be replaced by MCs without decreasing the efficiency of vector production and vector quality. MC-derived scAAV vector preparations even exceeded plasmid-derived preparations, as they displayed up to 30-fold improved transduction efficiencies. Using MCs as tools, we found that the vector plasmid is the main source of encapsidated prokaryotic sequences. Remarkably, we found that plasmid-derived scAAV vector preparations contained a much higher relative amount of prokaryotic sequences (up to 26.1 %, relative to TEC) compared to ssAAV vector preparations (up to 2.9 %). By replacing both plasmids by MCs the amount of functional prokaryotic sequences could be decreased to below the limit of quantification. Additional analyses for DNA impurities other than prokaryotic sequences showed that scAAV vectors generally contained a higher amount of non-vector DNA (e.g. adenoviral sequences) than ssAAV vectors. For both, ssAAV and scAAV vector preparations, MC-derived vectors tended to contain lower amounts of foreign DNA. None of the vectors tested could be shown to induce immunogenicity. In summary we could demonstrate that the quality of AAV vector preparations could be significantly improved by replacing producer plasmids by MCs. Upon transduction of a target tissue, AAV vector genomes predominantly remain in an episomal state, as duplex DNA circles or concatemers. These episomal forms mediate long-term transgene expression in terminally differentiated cells, but are lost in proliferating cells due to cell division. Therefore, in the second part of the thesis, in cooperation with Claudia Hagedorn and Hans J. Lipps (University Witten/Herdecke) an AAV vector genome was equipped with an autonomous replication element (Scaffold/matrix attachment region (S/MAR)). AAV-S/MAR encoding for eGFP and a blasticidin resistance gene and a control vector with the same TEC but lacking the S/MAR element (AAV-ΔS/MAR) were produced and transduced into highly proliferative HeLa cells. Antibiotic pressure was employed to select for cells stably maintaining the vector genome. AAV-S/MAR transduced cells yielded a higher number of colonies than AAV-ΔS/MAR-transduced cells. Colonies derived from each vector transduction were picked and cultured further. They remained eGFP-positive (up to 70 days, maximum cultivation period) even in the absence of antibiotic selection pressure. Interestingly, the mitotic stability of both AAV-S/MAR and control vector AAV-ΔS/MAR was found to be a result of episomal maintenance of the vector genome. This finding indicates that, under specific conditions such as the mild selection pressure we employed, “common” AAV vectors persist episomally. Thus, the S/MAR element increases the establishment frequency of stable episomes, but is not a prerequisite.

Normative reference values for the 20-meter shuttlerun test in a population-based sample of schoolchildren in Bogota, Colombia: The FUPRECOL Study

Our aim was to determine the normative reference values of cardiorespiratory fitness (CRF) and to establish the proportion of subjects with low CRF suggestive of future cardio-metabolic risk.

Life Design Counseling Group Intervention With Portuguese Adolescents

This article examines the process and outcome of a life design counseling group intervention with students in Grades 9 and 12. First, we applied a quasi-experimental methodology to analyze the intervention’s effectiveness in promoting career certainty, career decision-making, self-efficacy, and career adaptability in a sample of 236 students. Second, focus groups comprising 33 participants were conducted, examining participants’ perceptions of the intervention process and outcome. Our findings showed that the intervention had a significant effect on both career certainty and career self-efficacy, but it had no effect on career adaptability. Our results also showed that My Career Story (MCS) had a stronger effect on Grade 12 students. Focus group participants reported on the usefulness of MCS, as well as on its benefits, which include increased information as well as a sense of direction, self-discovery, connection, and increased self-awareness. Grade 9 participants expressed more difficulties in narrating self-experience than Grade 12 participants did. Implications for future research and practice are discussed.

Analisi multisensore di un evento temporalesco estremo nel Golfo di Napoli

In questo lavoro di tesi è stato analizzato il sistema convettivo a mesoscala (Mesoscale Convective System (MCS)) del 05/09/2015, abbattutosi sui golfi di Gaeta e di Napoli, sulla stessa città di Napoli e sull'entroterra irpino, scaricando una notevole quantità di fulmini e chicchi di grandine di dimensioni molto al di sopra di quelle usuali nella regione mediterranea, con un diametro compreso tra i 5 ed i 10 cm. Nel primo capitolo vengono presentati gli strumenti utilizzati per l'acquisizione dei dati da satellite, dei fulmini e delle mappe di previsione. Nel secondo capitolo viene effettuata un'analisi meteorologica, in cui vengono descritte le condizioni meteorologiche precedenti e contemporanee all'evento, col supporto di dati provenienti dai modelli numerici di previsione, dalle immagini da satellite e dai radiosondaggi; viene inoltre giudicata la performance di previsione dei modelli. Nel terzo capitolo viene descritto il lavoro di elaborazione dei dati da satellite e vengono esposti i risultati e i grafici ottenuti sull'evoluzione della nube. Si descrive l'elaborazione dei dati sui fulmini e, tramite mappe e grafici, si ricercano correlazioni e risultati. Nel quarto capitolo vengono mostrati alcuni fotogrammi estratti da video presenti su Internet che dimostrano l'eccezionalità dell'evento.

Interference mitigation techniques for cloud-RAN deployments in 5G and beyond systems

The deployment of ultra-dense networks is one of the most promising solutions to manage the phenomenon of co-channel interference that affects the latest wireless communication systems, especially in hotspots. To meet the requirements of the use-cases and the immense amount of traffic generated in these scenarios, 5G ultra-dense networks are being deployed using various technologies, such as distributed antenna system (DAS) and cloud-radio access network (C-RAN). Through these centralized densification schemes, virtualized baseband processing units coordinate the distributed access points and manage the available network resources. In particular, link adaptation techniques are shown to be fundamental to overall system operation and performance enhancement. The core of this dissertation is the result of an analysis and a comparison of dynamic and adaptive methods for modulation and coding scheme (MCS) selection applied to the latest mobile telecommunications standards. A novel algorithm based on the proportional-integral-derivative (PID) controller principles and block error rate (BLER) target has been proposed. Tests were conducted in a 4G and 5G system level laboratory and, by means of a channel emulator, the performance was evaluated for different channel models and target BLERs. Furthermore, due to the intrinsic sectorization of the end-users distribution in the investigated scenario, a preliminary analysis on the joint application of users grouping algorithms with multi-antenna and multi-user techniques has been performed. In conclusion, the importance and impact of other fundamental physical layer operations, such as channel estimation and power control, on the overall end-to-end system behavior and performance were highlighted.