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publicado em: Eur J Hum Gen.2016;24(S1):4. PL2.1 Balanced chromosomal abnormalities (BCAs) represent a unique class of genomic variation that involves large rearrangement of the chromosomes. To date their detection has been limited to cytogenetic resolution as most first-tier genetic screening methods are blind to their presence. We defined the genomic landscape of de novo BCAs associated with human congenial anomalies in 235 subjects using whole-genome sequencing. We observed that 22% of all BCAs harbored additional cryptic complexity, ranging from three breakpoints to chromothripsis events involving up to 57 breakpoints. Compared to random expectations, BCAs were more likely to occur between loci in close physical proximity in the nucleus, and their breakpoints were significantly enriched for evolutionarily constrained and embryonically expressed genes. From our convergent genomic interpretation using orthogonal datasets, we predict that the congenital anomaly phenotype was likely attributable to the BCA in at least 30% of subjects. An additional 4% of BCAs disrupted long-range regulatory regions such as topologically associating domains (TADs) resulting in position effects on genes associated with specific clinical manifestations that were compatible with the proband sequenced here. Remarkably, we observed a cluster of six independent translocations that disrupted a TAD and consequently altered MEF2C expression, mimicking the 5q14.3 microdeletion syndrome. These results suggest that de novo BCAs represent a highly penetrant class of genomic variation associated with congenital anomalies, and that nucleotide resolution offers insights into phenotypic prediction from direct gene disruption and alteration of long-range regulatory domains that are likely to be a significant source of causal variation in human disease. info:eu-repo/semantics/publishedVersion

Dissertação de mestrado integrado em Engenharia Biomédica (área de especialização em Engenharia Clínica), Na cavidade oral, as espécies de Candida co-existem com outras espécies de Candida e/ou bactérias. Nos últimos anos as infecções causadas por Candida têm aumentado, sendo Candida albicans o fungo mais isolado. No entanto, outras espécies como Candida parapsilosis têm emergido como agente patogénico e colonizador de superfícies orais, nomeadamente próteses dentárias. O trabalho desenvolvido teve como objectivo estudar diferentes factores de virulência nomeadamente adesão, formação de biofilme (biomassa total e produção da matriz extracelular) e expressão de enzimas hidrolíticas de isolados clínicos orais de C. albicans e C. parapsilosis em sistemas de mono-espécie e de duas-espécies quando aderidos a uma superfície de poliestireno revestida com saliva artificial. Os isolados clínicos orais utilizados neste estudo foram isolados em condições diferentes: C. albicans AM e C. parapsilosis AM2 isolados de coinfecção e C. albicans AC e C. parapsilosis AD isolados de mono-infecção. Neste trabalho, quando as espécies isoladas em co-infecção foram separadas verificou-se que ao fim de 2h de adesão, C. parapsilosis AM2 é influenciado pela ausência de C. albicans AM, necessitando de mais tempo para se adaptar ao novo ambiente. Todavia, C. albicans AM ao fim de 2h tem capacidade de se adaptar a novos ambientes na ausência de C. parapsilosis AM2, quer num sistema de mono-espécie e quer num sistema de duas-espécies com a estirpe C. parapsilosis AD. Em biofilme simples, as estirpes de C. parapsilosis apresentaram maior expressão de factores de virulência do que as estirpes de C. albicans. Em biofilme misto, a expressão de factores de virulência foi menor do que o somatório da expressão de cada espécie em biofilme simples. Verificou-se ainda que o sistema de duasespécies formado pelas espécies isoladas em co-infecção produziu mais biomassa total e matriz extracelular. Nos biofilmes mistos formados por uma espécie isolada em co-infecção e a outra espécie isolada em mono-infecção não se detectou a produção de polissacáridos da matriz extracelular, apesar de cada uma das espécies produzirem polissacáridos em biofilme simples. Neste trabalho também se verificou que é durante a fase de adesão que as espécies de Candida apresentam maior actividade enzimática, salientando-se que nos sistemas de duasespécies, na presença de C. parapsilosis AM2, a actividade enzimática foi maior do que na presença de C. parapsilosis AD. Assim, neste trabalho evidenciou-se que a expressão de factores de virulência depende das condições em que as espécies foram isoladas (mono-infecção ou co-infecção), da estirpe, do tipo de sistema (mono-espécie ou duas-espécies). A compreensão dos mecanismos de interacção entre as duas espécies poderá contribuir para o desenvolvimento de estratégias mais eficazes no controlo destes patogénicos oportunistas., In the oral cavity, Candida species co-exist with other Candida spp. and/or bacteria. In recent years, infections caused by Candida spp. have increased and Candida albicans is the most common fungal isolated. However, Candida parapsilosis is emerging as pathogen and colonizing oral surfaces, such as dental prosthesis. This work aimed to study the ability of oral clinical isolates of C. albicans and C. parapsilosis to express virulence factors such as adhesion, biofilm formation (total biomass and biofilm matrix composition) and secretion of hydrolytic enzymes in systems of single-species and dual-species, after adhesion onto an artificial saliva polystyrene coated surface. The clinical isolates used in this study were isolated from different conditions: C. albicans AM and C. parapsilosis AM2 were isolated in co-infection and C. albicans AC and C. parapsilosis AD were isolated in single-infection. In this study, when species isolated in co-infection were separated, it was verified that after 2h of adhesion, C. parapsilosis AM2 was influenced by the absence of C. albicans AM, needing more time to adapt to the new environment. However, C. albicans AM after 2h of adhesion has the ability to adapt to a new environment in the absence of C. parapsilosis AM2, either in mono-specie system and in dual-species system with C. parapsilosis AD. In simple biofilm, strains of C. parapsilosis had higher expression of virulence factors than strains of C. albicans. In mixed biofilm, the expression of virulence factors was lower than the sum of the expression of each species in simple biofilm. It was also verified that dual-species system formed by species isolated in co-infection produced more total biomass and extracellular matrix. In mixed biofilms formed by the specie isolated in co-infection and other specie isolated in mono-infection the production of polysaccharides was not detected, although each species in simple biofilm produce polysaccharides. It was also found that during adhesion Candida species have higher enzyme activity. The dual-species systems with C. parapsilosis AM2 have higher enzymatic activity than the dualspecies systems with C. parapsilosis AD. Thus, this study showed that expression of virulence factors is dependent on the condition from which the strains were isolated (single or co-infection), strain, type of culture (monospecie or dual-specie). The understanding of the interaction mechanisms between these species may contribute to a better control of their pathogenicity.

Os autores, após historiar a desccberta do Trypanosoma conorrhini (DONOVAN, 1909) (sinonimia Crithidia conorrhini DONOVAN, 1909, Trypanosoma boy lei LAFONT, 1912) no inseto transmissor e no hospedeiro vertebrado, o Rattus rattus diardi, fazem um estudo sumário do seu vector intermediário, o barbeiro cosmopolita Triatoma rubrofasciata (DE GEER, 1773) . Em seguida, referem a captura, no centro da cidade do Rio de Janeiro, Brasil, de um exemplar adulto dêste barbeiro parasitado por flagelados que foram identificados ao Trypanosoma conorrhini, fato êste pela primeira vez verificado no Novo Mundo. Certas formas evolutivas dêste parasito no inseto são muito semelhantes às do Schizotrypanum cruzi, mas os tripanosomas metacíclicos apresentam alguns caractéres morfológicos que permitem seu reconhecimento: os mais notáveis são o pequeno tamanho e a colocação do núcleo muito para trás, bem junto ao blefaroplasto. São dadas as medidas de 100 tripanosomas metaciclicos do T. conorrhini e de Schizotrypanum, cujo comprimento total médio foi, respectivamente, 13.88 u e 19.85 u. Nas seguintes espécies de barbeiro foi facilmente obtida a evolução do Trypanosoma conorrhini: Triatoma infestans, Triatoma vitticeps, Rhodnius prolixus e Panstrongylus megistus. Por inoculação de triatomas infectados foi obtida a transmissão do T. conorrhini ao camondongo branco, ao rato e ao macaco rhesus. As infecções foram muito fracas, custando-se a ver o tripanosoma no sangue, a fresco. Em gotas espêssas êle é encontrado com mais facilidade, mas o método de escôlha para verificação da sua presença no sangue dos animais é o xenodiagnóstico. Nesta prova o Triatoma infestans foi empregado com os resultados mais favoráveis. Por êste processo foi observada a infecção inaparente de um camondongo até 53 dias depois da inoculação. A forma sanguícola do T. conorrhini é um tripanosoma ttpico, de grande dimensões (40-60u), de extremidade posterior muito alongada e pontuda, membrana ondulante ampla e com largas pregas, blefaroplasto subcentral; em muitos indivíduos é notável uma estrutura particular, junto e à frente do blefaroplasto. Poucas horas depois de inoculados com as dejeções, os flagelados passam para a circulação, transformando-se em 3-4 dias nos grandes tripanosomas que acabam de ser descritos. O número de parasitos depende do número de flagelados inoculados. Não se conhece o processo de multiplicação do T. conorrhini no vertebrado, nunca tendo sido observados tripanosomas em via de divisão no sangue nem formas proliferativas nos tecidos. As sub-inoculações (de animal a animal) em geral não passam da primeira (MORISHITA, 1935). Tentativas até agora realizadas por outros pesquisadores no sentido de cultivar o T. conorrhini a partir do sangue de animais infectados, têm sido negativas. Por enquanto as pesquisas tendentes a determinar o hospedeiro vertebrado do parasito no Rio de Janeiro limitam-se a aplicação do xenodiagnós-tico em cinco ratos do Morro de Santo Antônio, cujo resultado... In the present paper details are given on the discovery of Trypanosoma conorrhini (DONOVAN, 1909) (syn. Crithidia conorrhini DONOVAN, 1909; Trypanosoma boy lei LAFONT, 1912) in its insect vector, Triatoma rubrofasciata (DE GEER, 1773) and in its vertebrate host, the rat (Rattus rattus diardi). The universal geographic distribution of Triatoma rubrofasciata is recorded. The writers report the finding in Rio de Janeiro, Brazil, of an adult example of T. rubrofasciata infected with flagellates that are referable to Trypanosoma conorrhini. The close resemblance of certain forms of this flagellate to the insectan forms of Schizotrypanum cruzi is emphasized. Metacyclic trypanosome forms of each species, however, may be distinguished; those of Trypanosoma conorrhini are shorter and their nucleus is closer to the parabasal body than in Schizotrypanum cruzi. Data concerning the measures of 100 metacyclic forms of each species are given: The mean total length of T. conorrhini was 13.88U and that of Schizotrypanum was 19.85µ. The experimental evolution of T. conorrhini has been easily obtained in Triatoma infestans, Triatoma vitticeps, Rhodnius prolixus and Panstrongylus megistus. By the intraperitoneal inoculation of white mouse, rat and Rhesus monkey light infection were produced. Parasites were rarely seen in fresh blood preparations, but could be easily demonstrated in the peripheral blood of apparently recovered mice untill 53 days after the inoculation, by "xenodiagnosis". The blood form of Trypanosoma conorrhini is a large trypanosome with long and pointed posterior end, a sub-central parabasal body with a particular pot-shaped structure just in front of it, and a wide ondulating membrane. A few hours after the peritoneal inoculation of the gut contents of infected Triatoma rubrofasciata parasites may appear in the blood, where in 3-4-days they reach the large size of the typical trypanosome form. The number of trypanosomes in the peripheral blood depends upon the number of parasites in the inoculum. It is interesting to note that so far division forms of T. conorrhini have never been found in the blood or tissues of infected animals. Sub-inoculation of the blood forms usually do not succeeded beyond the first, and even the first sub-inoculation result in very light infection. Cultures from the infected animal blood have not yet succeded (MORISHITA, 1935). So far Trypanosoma conorrhini appear to be the only species of Trypa-nosoma transmitted by reduviid-bugs; a list is given of trypanosomes which failed to develop in such bugs on experimental conditions.

We describe two new missense mutations in the glucose-6-phosphate dehydrogenase (G6PD) gene associated with chronic hemolytic anemia: mutation 1205CÆÆA in exon 10 predicts the amino acid change 402ThrÆÆAsn in the bb-sheet M of the polypeptide chain, within the dimer interface (G6PD Covão do Lobo); mutation 1366GÆÆA in exon 12 predicts the amino acid substitution 456AspÆÆHis in the aa-helix N, at the protein surface (G6PD Figueira da Foz).

Companies have recognized the importance of viral marketing that has an increasing role in the marketing world because in this interactive process consumers may easily and quickly connect with companies and other consumers ; they can exchange information about products and have complete freedom in the transfer of experience. On the other hand, companies can directly approach to customers, get feedback regarding their satisfaction and thus create a long-term relationship with them. Regardless of the technology and the internet that change the habits of consumers and strongly influence the buying decision, the effect of word-of-mouth and recommendations of people who are close to the customers will always be significant. The purpose of this paper is to compare the viral marketing and word-of-mouth, two very important marketing communication channels that function very similarly, but at the same time use different techniques to reach out the customers. This paper attempts to compare the impact of viral marketing and word-of-mouth when making decisions about buying and to create awareness of the brand and strengthen the brand image through research.

Innovations are the foundation of progress, especially innovations in fields with fast and dynamic cycles. The purpose of this paper is to give an overview of key elements in implementation of new marketing and communication channels strategy. The paper demonstrate that new and innovative marketing tools contribute to the overall success of small and medium-sized companies ; these often have limited budgets and therefore have to be smarter in selecting the ways to use them. The basis for this analysis is an overview of available literature. Diversity and relevance of the source matter for the overall quality of the work and its practical application. First, the present status of the tools and channels is analysed, then predictions are made about changes in a company after the introduction of the proposed changes in marketing and communication tools.

Nautical tourism is the most perspective industry in the Republic of Croatia. Its growth and development in the last ten years made nautical tourism one of the most competitive and profitable „tourist product“. The paper analyzes and evaluates current state of the nautical tourism in the Republic of Croatia with specific references to the management of the nautical ports. The aim of the paper is to indicate a high level of development and further growth of nautical tourism and to attempt to provide an answer if the nautical tourism has a significant and economic impact on the gravitating destination. The profit from nautical tourism is not commensurate to its potentials. Analyzing current state and dynamics of its growth we can define some basic regulations, which are indicating that its significant growth is still to expect. The results revealed constant growth of nautical tourism and its major impact on destination development.