Characterization of nanoparticle mediated laser transfection by femtosecond laser pulses for applications in molecular medicine

Background: In molecular medicine, the manipulation of cells is prerequisite to evaluate genes as therapeutic targets or to transfect cells to develop cell therapeutic strategies. To achieve these purposes it is essential that given transfection techniques are capable of handling high cell numbers in reasonable time spans. To fulfill this demand, an alternative nanoparticle mediated laser transfection method is presented herein. The fs-laser excitation of cell-adhered gold nanoparticles evokes localized membrane permeabilization and enables an inflow of extracellular molecules into cells. Results: The parameters for an efficient and gentle cell manipulation are evaluated in detail. Efficiencies of 90% with a cell viability of 93% were achieved for siRNA transfection. The proof for a molecular medical approach is demonstrated by highly efficient knock down of the oncogene HMGA2 in a rapidly proliferating prostate carcinoma in vitro model using siRNA. Additionally, investigations concerning the initial perforation mechanism are conducted. Next to theoretical simulations, the laser induced effects are experimentally investigated by spectrometric and microscopic analysis. The results indicate that near field effects are the initial mechanism of membrane permeabilization. Conclusion: This methodical approach combined with an automated setup, allows a high throughput targeting of several 100,000 cells within seconds, providing an excellent tool for in vitro applications in molecular medicine. NIR fs lasers are characterized by specific advantages when compared to lasers employing longer (ps/ns) pulses in the visible regime. The NIR fs pulses generate low thermal impact while allowing high penetration depths into tissue. Therefore fs lasers could be used for prospective in vivo applications.

Russian social networks on the web: cohesion and coherence in Vkontakte

In this thesis connections between messages on the public wall of the Russian social network Vkontakte are analysed and classified. A total of 1818 messages from three different Vkontakte groups were collected and analysed according to a new framework based on Halliday and Hasan’s (1976) research into cohesion and Simmons’s (1981) adaptation of their classification for Russian. The two categories of textuality, cohesion and coherence, describe the linguistic connections between messages. The main aim was to find out how far the traditional categories of cohesion are applicable to an online social network including written text as well as multimedia-files. In addition to linguistic cohesion the pragmatic and topic coherence between Vkontakte messages was also analysed. The analysis of pragmatic coherence classifies the messages with acts according to their pragmatic function in relation to surrounding messages. Topic coherence analyses the content of the messages, describes where a topic begins, changes or is abandoned. Linguistic cohesion, topic coherence and pragmatic coherence enable three different types of connections between messages and these together form a coherent communication on the message wall. The cohesion devices identified by Halliday and Hasan and Simmons were found to occur in these texts, but additional devices were also identified: these are multimodal, graphical and grammatical cohesion.

Prosument Behaviour In Brand Creating

A brand remains a considerable source of the competitive advantage. One of the elements contributing to its power is image. The information revolution and globalization make it necessary to search for new means of differentiating brands. One of them is engaging consumers in the brand creation process. In light of the development of the Web 2.0, prosumers – active consumers functioning both as consumers and partly as producers – can have a meaningful influence on the image of brands. Their activities can entail both positive as well as negative effects.

Navegar na web : as redes sociais como espaços de comunicação, partilha de conhecimento e de promoção de competências de aprendizagem

A aprendizagem em rede e as potencialidades do software social trouxeram novos e estimulantes desafios para os sistemas educativos e para os seus profissionais. Um dos principais desafios prende-se com a necessidade de conceber uma “nova” didática para a docência na web social que deve basear-se não só nos conhecimentos científico, tecnológico, curricular e pedagógico, mas também num conhecimento científico e pedagógico da tecnologia que permita planear, conceber e utilizar as redes sociais, como o Facebook, no processo de ensino-aprendizagem de forma eficaz. Assim, e perante esta realidade, neste estudo exploratório procurámos perceber em que medida a utilização do Facebook no processo de ensino-aprendizagem permite a promoção de competências de aprendizagem de estudantes de cursos pós-graduados, a nível da capacidade para aprender, da iniciativa e da autonomia. Os resultados sugerem que os estudantes aceitam o Facebook como um novo contexto para a aprendizagem, que não impede a reflexão crítica sobre os conceitos e as temáticas apresentadas para trabalho, possibilitando o desenvolvimento de comunidades de aprendizagem desde que exista uma intencionalidade educativa explícita.

A novel concept-level approach for ultra-concise opinion summarization

The Web 2.0 has resulted in a shift as to how users consume and interact with the information, and has introduced a wide range of new textual genres, such as reviews or microblogs, through which users communicate, exchange, and share opinions. The exploitation of all this user-generated content is of great value both for users and companies, in order to assist them in their decision-making processes. Given this context, the analysis and development of automatic methods that can help manage online information in a quicker manner are needed. Therefore, this article proposes and evaluates a novel concept-level approach for ultra-concise opinion abstractive summarization. Our approach is characterized by the integration of syntactic sentence simplification, sentence regeneration and internal concept representation into the summarization process, thus being able to generate abstractive summaries, which is one the most challenging issues for this task. In order to be able to analyze different settings for our approach, the use of the sentence regeneration module was made optional, leading to two different versions of the system (one with sentence regeneration and one without). For testing them, a corpus of 400 English texts, gathered from reviews and tweets belonging to two different domains, was used. Although both versions were shown to be reliable methods for generating this type of summaries, the results obtained indicate that the version without sentence regeneration yielded to better results, improving the results of a number of state-of-the-art systems by 9%, whereas the version with sentence regeneration proved to be more robust to noisy data.

O marketing das notícias : consumo de informações online, persuasão e confiança

Dissertação (mestrado)—Universidade de Brasília, Instituto de Psicologia, Departamento de Psicologia Social e do Trabalho, Programa de Pós-Graduação em Psicologia Social, do Trabalho e das Organizações, 2016.

SECURITY, PRIVACY AND APPLICATIONS IN VEHICULAR AD HOC NETWORKS

With wireless vehicular communications, Vehicular Ad Hoc Networks (VANETs) enable numerous applications to enhance traffic safety, traffic efficiency, and driving experience. However, VANETs also impose severe security and privacy challenges which need to be thoroughly investigated. In this dissertation, we enhance the security, privacy, and applications of VANETs, by 1) designing application-driven security and privacy solutions for VANETs, and 2) designing appealing VANET applications with proper security and privacy assurance. First, the security and privacy challenges of VANETs with most application significance are identified and thoroughly investigated. With both theoretical novelty and realistic considerations, these security and privacy schemes are especially appealing to VANETs. Specifically, multi-hop communications in VANETs suffer from packet dropping, packet tampering, and communication failures which have not been satisfyingly tackled in literature. Thus, a lightweight reliable and faithful data packet relaying framework (LEAPER) is proposed to ensure reliable and trustworthy multi-hop communications by enhancing the cooperation of neighboring nodes. Message verification, including both content and signature verification, generally is computation-extensive and incurs severe scalability issues to each node. The resource-aware message verification (RAMV) scheme is proposed to ensure resource-aware, secure, and application-friendly message verification in VANETs. On the other hand, to make VANETs acceptable to the privacy-sensitive users, the identity and location privacy of each node should be properly protected. To this end, a joint privacy and reputation assurance (JPRA) scheme is proposed to synergistically support privacy protection and reputation management by reconciling their inherent conflicting requirements. Besides, the privacy implications of short-time certificates are thoroughly investigated in a short-time certificates-based privacy protection (STCP2) scheme, to make privacy protection in VANETs feasible with short-time certificates. Secondly, three novel solutions, namely VANET-based ambient ad dissemination (VAAD), general-purpose automatic survey (GPAS), and VehicleView, are proposed to support the appealing value-added applications based on VANETs. These solutions all follow practical application models, and an incentive-centered architecture is proposed for each solution to balance the conflicting requirements of the involved entities. Besides, the critical security and privacy challenges of these applications are investigated and addressed with novel solutions. Thus, with proper security and privacy assurance, these solutions show great application significance and economic potentials to VANETs. Thus, by enhancing the security, privacy, and applications of VANETs, this dissertation fills the gap between the existing theoretic research and the realistic implementation of VANETs, facilitating the realistic deployment of VANETs.

Humanidades digitales y la escuela 2.0

During the last decades the growth and development of Information and Communication Technologies (ICT) have led us to a new social paradigm that reflects a deep change not only in an individual but also in a social behavioural pattern. All these changes define the so-called knowledge and information society. This social evolution has had different textual states and types as its main core and instrument of transformation therefore scholars and specialists in the field of Humanities have not missed the opportunity of studying recent phenomena, which have come from the current setting given by new technologies. Researchers in Humanities have had to reconsider their own traditional working method based on printed text in order to analyse how nowadays we search for data, select them, analyse them and the ways we create and spread new information and knowledge. Having in mind this scenario where humanist works, the concept of Digital Humanities has arisen conditioned by the existence of cyberspace, digital text, hypertext, on-line text, implementation of new ways of communication, global access to information and various elements which build a common methodology for all humanistic disciplines. Most of these changes have affected the educational system and education as an academic and humanistic discipline.

The actinin family of actin crosslinking proteins: natural functions and potential applications in synthetic biology

Actinin and spectrin proteins are members of the Spectrin Family of Actin Crosslinking Proteins. The importance of these proteins in the cytoskeleton is demonstrated by the fact that they are common targets for disease causing mutations. In their most prominent roles, actinin and spectrin are responsible for stabilising and maintaining the muscle architecture during contraction, and providing shape and elasticity to the red blood cell in circulation, respectively. To carry out such roles, actinin and spectrin must possess important mechanical and physical properties. These attributes are desirable when choosing a building block for protein-based nanoconstruction. In this study, I assess the contribution of several disease-associated mutations in the actinin-1 actin binding domain that have recently been linked to a rare platelet disorder, congenital macrothrombocytopenia. I investigate the suitability of both actinin and spectrin proteins as potential building blocks for nanoscale structures, and I evaluate a fusion-based assembly strategy to bring about self-assembly of protein nanostructures. I report that the actinin-1 mutant proteins display increased actin binding compared to WT actinin-1 proteins. I find that both actinin and spectrin proteins exhibit enormous potential as nano-building blocks in terms of their stability and ability to self-assemble, and I successfully design and create homodimeric and heterodimeric bivalent building blocks using the fusion-based assembly strategy. Overall, this study has gathered helpful information that will contribute to furthering the advancement of actinin and spectrin knowledge in terms of their natural functions, and potential unnatural functions in protein nanotechnology.

Computational characterization of carbazole-benzonitrile derivatives for applications in Organic Light Emitting Diodes

The technology of Organic Light-Emitting Diodes has reached such a high level of reliability that it can be used in various applications. The required light emission efficiency can be achieved by transforming the triplet excitons into singlet states through Reverse InterSystem Crossing (RISC), which is the main process of a general mechanism called thermally activated delayed fluorescence (TADF). In this thesis, we theoretically analyzed two carbazole-benzonitrile (donor-acceptor) derivatives, 2,5-di(9H-carbazol-9-yl)benzonitrile (p-2CzBN) and 2,3,4,5,6-penta(9H-carbazol-9-yl)benzonitrile (5CzBN), and addressed the problem of how donor-acceptor (D-A) or donor-acceptor-donor (D-A-D) flexible molecular architectures influence the nature of the excited states and the emission intensity. Furthermore, we analyzed the RISC rates as a function of the conformation of the carbazole lateral groups, considering the first electronic states, S0, S1, T1 and T2, involved in TADF process. The two prototype molecules, p-2CzBN and 5CzBN, have a similar energy gap between the first singlet and triplet states (∆EST, a key parameter in the RISC rate), but different TADF performances. Therefore, other parameters must be considered to explain their different behavior. The oscillator strength of p-2CzBN, never tested as emitter in OLEDs, is similar to that of 5CzBN, which is an active TADF molecule. We also note that the presence of a second T2 triplet state, lower in energy than S1 only in 5CzBN, and the reorganization energies, associated with RISC processes involving T1 and T2, are important factors in differentiating the rates in p-2CzBN and 5CzBN. For p-2CzBN, the RISC rate from T2 to S1 is surprisingly higher than that from T1 to S1, in disagreement with El-Sayed rules, due to a large reorganization energy associated to the T1 to S1, process; while the contrary occurs for 5CzBN. These insights are important for designing new TADF emitters based on the benzo-carbazole architecture.

Novel hydroxystearoyl-, heterocyclic and carbocyclic derivatives: synthesis and applications in biological field

This PhD project has been mainly focused on the synthesis of novel organic compounds containing heterocyclic and/or carbocyclic scaffold and on the study of stearic acid derivatives and their applications in biological field. The synthesis of novel derivatives of 9-hydroxystearic acid (9-HSA) evidenced how the presence of substituents on C9, able to make hydrogen bonds is of crucial importance for the biological activity. Also the position of the hydroxy group along the chain of hydroxystearic acids was investigated: regioisomers with the hydroxy group bound to odd carbons resulted more active than those bearing the hydroxy group on even carbons. Further, the insertion of (R)-9-HSA in magnetic nanoparticles gave a novel material which characterization remarked its suitability for drug delivery. Structural hybrids between amino aza-heterocycles and azelaic acid have been synthesized and some of them showed a selective activity towards osteosarcoma cell line U2OS. Several Apcin analogues bearing indole, benzothiazole, benzofurazan moieties connected to tryptaminyl-, amino pyridinyl-, pyrimidinyl- and pyrazinyl ring through a 1,1,1-trichloroethyl group were synthesized. Biological tests showed the importance of both the tryptaminyl and the pyrimidinyl moieties, confirming the effectiveness against acute leukemia models. The SNAr between 2-aminothiazole derivatives and 7-chlorodinitrobenzofuroxan revealed different behaviour depending from amino substituent of the thiazole. The reaction with 2-N-piperidinyl-, 2-N-morpholinyl-, or 2-N-pyrrolidinyl thiazole gave two isomeric species derived from the attack on C-5 of thiazole ring. Thiazoles substituted with primary- or not-cyclic secondary amines reacted with the exocyclic amino nitrogen atom giving a series of compounds whose biological activity have highlighted as they might be promising candidates for further development of antitumor agents. A series of 9-fluorenylidene derivatives, of interest in medical and optoelectronic field as organic scintillators, was synthesized through Wittig or Suzuky reaction and will be analyzed to test their potential scintillatory properties.

Nuovi modelli di business nel Web 3.0

Il Web 3.0 è un'estensione del web come lo conosciamo oggi basata sulla blockchain e che ha permesso negli ultimi anni lo sviluppo della Token Economy. I token e gli smart contract sono gli strumenti che permettono di creare nuovi modelli di business tokenizzati. Questa tesi ha lo scopo di capire quale sia il legame tra lo sviluppo delle tecnologie del Web 3.0 e i modelli di business che ne derivano, andando a individuare i modelli imprenditoriali più noti, la loro value proposition e le complicazioni ad essi associate.

Analisi e implementazione di un sistema di autorizzazione compatibile con token OAuth 2.0 e certificati VOMS

Il Worldwide LHC Computing Grid (WLCG) è una collaborazione internazionale costituita da decine di centri di calcolo distribuiti globalmente, la cui missione consiste nell'elaborazione delle grandi quantità di dati prodotti dai maggiori esperimenti di Fisica delle Alte Energie, in particolare quelli al CERN di Ginevra. Uno di questi centri di calcolo è ospitato presso il CNAF dell'Istituto Nazionale di Fisica Nucleare a Bologna, che contribuisce anche allo sviluppo di middleware per la gestione dell'infrastruttura. Molti componenti di tale middleware, che hanno funzionalità e scopi diversi, richiedono un servizio di autorizzazione versatile e compatibile con i meccanismi di autenticazione attualmente in uso, basati su token OAuth 2.0 e su certificati VOMS Proxy. In questa tesi si analizzerà l'architettura e l'implementazione di un proof-of-concept di un sistema di autorizzazione che soddisfi queste necessità, enfatizzando l'integrazione delle tecniche di autenticazione citate. Per dimostrare la sua versatilità, verrà illustrato il processo di interfacciamento con un componente middleware attualmente in sviluppo al CNAF. Il risultato finale ottenuto è un sistema che rispetta i vincoli richiesti e si integra facilmente con servizi eterogenei.

Microbial Evaluation Of Traumatized Teeth Treated With Triple Antibiotic Paste Or Calcium Hydroxide With 2% Chlorhexidine Gel In Pulp Revascularization.

Revascularization outcome depends on microbial elimination because apical repair will not happen in the presence of infected tissues. This study evaluated the microbial composition of traumatized immature teeth and assessed their reduction during different stages of the revascularization procedures performed with 2 intracanal medicaments. Fifteen patients (7-17 years old) with immature teeth were submitted to the revascularization procedures; they were divided into 2 groups according to the intracanal medicament used: TAP group (n = 7), medicated with a triple antibiotic paste, and CHP group (n = 8), dressed with calcium hydroxide + 2% chlorhexidine gel. Samples were taken before any treatment (S1), after irrigation with 6% NaOCl (S2), after irrigation with 2% chlorhexidine (S3), after intracanal dressing (S4), and after 17% EDTA irrigation (S5). Cultivable bacteria recovered from the 5 stages were counted and identified by means of polymerase chain reaction assay (16S rRNA). Both groups had colony-forming unit counts significantly reduced after S2 (P < .05); however, no significant difference was found between the irrigants (S2 and S3, P = .99). No difference in bacteria counts was found between the intracanal medicaments used (P = .95). The most prevalent bacteria detected were Actinomyces naeslundii (66.67%), followed by Porphyromonas endodontalis, Parvimonas micra, and Fusobacterium nucleatum, which were detected in 33.34% of the root canals. An average of 2.13 species per canal was found, and no statistical correlation was observed between bacterial species and clinical/radiographic features. The microbial profile of infected immature teeth is similar to that of primarily infected permanent teeth. The greatest bacterial reduction was promoted by the irrigation solutions. The revascularization protocols that used the tested intracanal medicaments were efficient in reducing viable bacteria in necrotic immature teeth.