Inhibition of hepatitis C virus RNA replication by ISG15 does not require its conjugation to protein substrates by the HERC5 E3 ligase

Chronic infection of the liver by hepatitis C virus (HCV) induces a range of host factors including IFN-stimulated genes such as ISG15. ISG15 functions as an antiviral factor that limits virus replication. Previous studies have suggested that ISG15 could influence HCV replication in both a positive and a negative manner. In this report, we determined the effect of ISG15 on HCV RNA replication in two independent cell lines that support viral genome synthesis by inhibiting ISG15 expression through small interfering RNA, short-hairpin RNA and CRISPR/Cas9 gene knockout approaches. Our results demonstrated that ISG15 impairs HCV RNA replication in both the presence and absence of IFN stimulation, consistent with an antiviral role for ISG15 during HCV infection. ISG15 conjugation to protein substrates typically requires the E3 ligase, HERC5. Our results showed that the inhibitory effect of ISG15 on HCV RNA replication does not require its conjugation to substrates by HERC5

A single amino acid substitution in the novel H7N9 influenza A virus NS1 protein increases CPSF30 binding and virulence

Although an effective interferon antagonist in human and avian cells, the novel H7N9 influenza virus NS1 protein is defective at inhibiting CPSF30. An I106M substitution in H7N9 NS1 can restore CPSF30 binding together with the ability to block host gene expression. Furthermore, a recombinant virus expressing H7N9 NS1-I106M replicates to higher titers in vivo, and is subtly more virulent, than parental. Natural polymorphisms in H7N9 NS1 that enhance CPSF30 binding may be cause for concern

Co-limitation of phytoplankton by N and P in a shallow coastal lagoon (Ria Formosa): implications for eutrophication evaluation

The Ria Formosa coastal lagoon is a highly productive shallow ecosystem in southern Portugal, subjected to nutrient inputs from anthropogenic and natural sources. Nutrients are major abiotic drivers of phytoplankton in this system, but their effects on phytoplankton assemblages and the occurrence of nutrient limitation are still poorly understood. The main goal of this study was, thus, to evaluate the occurrence, type, and effects of nutrient limitation on phytoplankton community and specific functional groups in the Ria Formosa coastal lagoon. We conducted nutrient enrichment experiments with factorial additions of nitrogen (N) and phosphorus (P) using natural phytoplankton assemblages from distinct locations in the Ria Formosa, throughout a yearly cycle. Phytoplankton composition and abundance were evaluated using inverted and epifluorescence microscopies, and spectrophotometric methods were used for biomass. Limitation was defined as higher phytoplankton growth following enrichment with a particular nutrient in relation to the non-enriched control. The most common type of phytoplankton limitation was simultaneous co-limitation by N and P; diatoms, as r-strategists, were the most frequently limited group. Single N and P limitation, and serial P limitation were also observed, as well as negative responses to nutrient enrichment. Group-specific responses to nutrient enrichment were not reflected in the relative abundance of phytoplankton groups within the whole assemblage, due to the numerical dominance of pico-sized groups (cyanobacteria and eukaryotic picophytoplankton). Ambient nutrient ratios and concentrations did not predict phytoplankton nutrient limitation, given the different nutrient utilisation traits among phytoplankton functional groups. Therefore, nutrient ratios should not be used as indicators of nutrient limitation in eutrophication assessment.LA/P/0069/2020info:eu-repo/semantics/publishedVersio

Investigação de componentes específicos e essenciais para exportação de mRNA de tripanossomatídeos

Orientadora : Dra. Andréa Rodrigues ÁvilaCoorientador : Prof. Dr. Alexandre Haruo InoueDissertação (mestrado) - Universidade Federal do Paraná, Setor de Ciências Biológicas, Programa de Pós-Graduação em Biologia Celular e Molecular. Defesa: Curitiba, 23/02/2017Inclui referências : f. 151-158Resumo: Trypanosoma cruzi é o protozoário causador da doença de Chagas, uma endemia da América Latina que afeta milhões de pessoas. Este parasita tem um ciclo de vida alternado entre hospedeiros vertebrados e invertebrados, e necessita se adaptar a mudanças em diferentes ambientes que encontra ao longo do ciclo de transmissão. O T. cruzi controla a expressão de genes a partir de eventos majoritariamente pós-transcricionais. Vias que controlam a estabilidade do mRNA ou a tradução de proteínas são bem conhecidas, enquanto que mecanismos moleculares de exportação de mRNA do núcleo para o citoplasma ainda não são bem compreendidos nem em tripanossomatídeos, nem em outros patógenos. Sabe-se, até o momento, que a maioria das proteínas envolvidas com o transporte de mRNA em outros eucariotos não está conservada em diversas espécies de parasitas. Exceto pela proteína Sub2 de T. cruzi (TcSub2), homóloga das proteínas de mamífero e levedura (UAP56/Sub2) e essencial para exportação de mRNA em tripanossomatídeos. Logo, para investigar os componentes desta via, nosso grupo tem investido em identificar proteínas exclusivas e essenciais para a exportação de mRNA em T. cruzi. Abordagens de proteômica, usando como alvo TcSub2, vem permitindo a identificação de fatores com função ainda desconhecida e/ou que são conservados apenas entre espécies de tripanossomatídeos, tornando-os alvos de interesse. Assim, o objetivo deste trabalho foi avaliar a interação de proteínas alvo com TcSub2 e seu papel na exportação de mRNA em T.cruzi. Duas proteínas não caracterizadas apresentaram motivos conservados e foram denominadas como TcFOP-like e TcAPI5-like. Interessantemente, estes motivos estão presentes em proteínas de outros organismos que estão envolvidas com a via de exportação de mRNA. Aqui, nós confirmamos que TcFOP-like e TcAPI5-like são proteínas nucleares e parecem interagir com TcSub2. Além disso, silenciamento e supereexpressão de TcFOP-like afeta o crescimento do parasita, e nós confirmamos sua interação com outros componentes da via de exportação de mRNA, tais como TcMex67 e TcHel45. Estas evidências suportam presença de components diveregentes na via de exportação de mRNA em Tripanossomatídeos. Palavras chaves: Trypanosoma cruzi. Exportação de mRNA. TcSub2.TcHel45Abstract: Trypanosoma cruzi is the protozoan causative of Chagas disease, an endemic disease in Latin America that affects millions of people. This parasite alternates between vertebrate and invertebrate hosts during his life cycle, and needs to adapt to the changes in different environments along the transmission cycle. T. cruzi controls the gene expression mostly by post-transcriptional events. Pathways that control mRNA stability or protein translation are well known, while the molecular mechanisms for mRNA export from nucleus to cytoplasm are still not well understood nether in trypanosomatids or other pathogens. So far, it's known that the most of the proteins that are involved in mRNA transport of other eukaryotes are not conserved in many species of parasites. One exception is T. cruzi Sub2 (TcSub2), a homologue of the proteins in mammalian and yeast (UAP56 / Sub2) and essential for mRNA export in tripanossomatids. Therefore, to investigate the components of this pathway, our group has invested in identifying unique and essential proteins for the export of mRNA in T. cruzi. Proteomic approaches, using TcSub2 as target, have allowed identification of factors with the unknown function and/or that are conserved only in species of trypanosomatids, turning into interesting targets. Then, the aim of this work was to evaluate the interaction of target proteins with TcSub2 and their role in the mRNA export pathway in T. cruzi. Two uncharacterized proteins presented conserved motifs and were named here as TcFOP-Like and TcAPI5-like. Interestingly, those motifs are present in proteins of other organisms which are involved in mRNA export pathway. Here, we confirmed that TcFOP-like and TcAPI5-like are nuclear proteins and seems to interact with TcSub2. Furthemore, knockdown and overexpression of TcFOP-like affect the growth of parasites and we confirmed its interaction with other components of the mRNA export pathway, such as TcMex67 and TcHel45. Those evidences supporte the presence of divergent components in the mRNA export pathway of Tripanossomatids. Keywords: Trypanosoma cruzi. Export of mRNA. TcSub2. TcHel45

Influenciadores digitais como fonte de valor das marcas: quem disse, Berenice?: um estudo de caso

Os influenciadores digitais são uma realidade do séc. XXI, na qual as marcas deixam de transmitir as mensagens em nome próprio e passam a usar uma pessoa, um influenciador, detentor de um poder de influência sobre uma comunidade online. Assim, as marcas fazem uso de uma comunidade já existente e do líder de opinião que vai desencadear um feedback positivo por parte destes. O objetivo do presente estudo é entender, através de uma análise descritivo-analítica, de que forma é que as publicações feitas por um influenciador geram electronic word-of-mouth. Pretende-se também perceber, do ponto de vista de uma marca, de que forma é que os influenciadores digitais podem ser responsáveis pela geração de valor para a marca. Assim, e de forma particular, o estudo foca-se em 10 publicações em que a marca “quem disse, berenice?” fez uso de influenciadores, para transmitir as mensagens pretendidas. O estudo analisa cada uma das publicações, o eWOM de cada um deles e de que forma é que isto pode contribuir para o valor da marca. O estudo observa de que forma é que a personalidade das influenciadoras está presente nas publicações que fazem, a relação que essa personalidade tem com a personalidade da marca e de que forma é que elas conseguem criar associações entre marca-seguidores e gerar resposta por parte destes últimos

Control of hepatitis C virus genome replication by viral factors and mediators of host intrinsic immunity

No abstract available

Control software for processing of materials and devices by ultraviolet laser

Orientadores: Jacobus Willibrordus Swart, Ednan JoanniDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia Elétrica e de ComputaçãoResumo: É implementado um conjunto de software de controle para modificação de superfícies e a fabricação de componentes através da incidência de um laser ultravioleta (UV) pulsado. Para tal é necessário realizar o controle sincronizado (por software dedicado) de uma controladora de movimentos com sistema de tempo real VxWorks, de posicionadores de alta precisão e de um laser UV de Nd:YVO4. A linguagem de programação escolhida foi Python. A finalidade do desenvolvimento do software é a utilização para o processamento de materiais, dispositivos e o tratamento de superfícies com o laser. Dentre os materiais utilizados estão: silício, vidros, alumina e polímeros. É necessário o estudo das condições ideais de trabalho para cada tipo de substrato. O software viabiliza a utilização dos componentes de forma integrada e precisa, permitindo a realização de trabalhos de microusinagem a laser, por processamento de imagens e via execução de trajetórias. Dentre as aplicações realizadas através do software implementado estão: o inchamento de polímeros (utilizado para a construção de moldes de microfluídica); o corte de células fotovoltaicas para produção de painéis customizados de energia solar; a produção de máscaras de sombra em vidro para deposição por sputtering de filmes finos e nanoestruturas em regiões pré-definidas; a escrita direta de eletrodos em substratos de alumina com deposição de ouro ou platina, para sensores de gásAbstract: It is implemented a set of control software for surface modification and manufacturing components via incidence of an ultraviolet laser (UV) pulsed. This requires performing a synchronized control (for dedicated software) of a motion controller with Real Time Operating System VxWorks, high performance positioners and a UV laser Nd: YVO4. The chosen programming language is Python. The purpose of software development is the use for processing of materials, devices and surface treatment with the laser. These materials are used: silicon, glasses and polymers. It is necessary to study an ideal working conditions for each type of substrate. The software enables the use of components in an integrated and precise manner, allowing the performance of work of laser micromachining, for image processing and track running paths. Among the applications performed by the software are implemented: the swelling polymer (used for the construction of microfluidic molds); cutting photovoltaic cells to produce customized solar panels; the production of shadow masks in glass deposition by sputtering of thin films and nanostructures on predefined regions; direct writing electrodes alumina substrates with deposition of gold or platinum, for gas sensorsMestradoEletrônica, Microeletrônica e OptoeletrônicaMestra em Engenharia Elétrica133971/2014-7CNP

Multi-Mode adhesives performance and success/retention rates in NCCLs restorations: randomised clinical trial one-year report

Aim: Compare clinical performance and success/retention rates of two multi-mode (MM) adhesives, applied in self-etch (SE) or etch-and-rinse (ER) modes, with SE-all-in-one adhesive (SE/SE with enamel etching) in NCCL restorations at one-year follow-up. Material and methods: Prospective, double-blind RCT approved by the University Fernando Pessoa and the National-Clinical-Research-Ethics Committees (CEIC-20150305), ClinicalTrials.gov registered (NCT02698371), in 38 participants with 210 restorations (AdmiraFusionVR ) randomly allocated to six groups (Adhesives_Adhesion mode), each with 35 restorations: G1-Control FuturabondVRDC_SE; G2-Control FuturabondVRDC_SE with enamel etching; G3-FuturabondVRU_ER; G4-FuturabondVR U_SE; G5-AdheseVR Universal_ER; G6-AdheseVR Universal_SE. Restorations evaluated at baseline and one-year by three calibrated examiners (ICC 0.952) using FDI criteria and statistical analysis with nonparametric tests (alpha=0.05). Results: At one-year recall 36 participants, 199 restorations were available for examination; five (2.5%) restorations (G1 n=2; G2, G3, G4 n=1) were lost due to retention (p>.05); G1 showed less satisfying marginal adaptation (p<.05) than G2 and MM adhesives groups, particularly G6. Overall success rates (p>.05) were: 93.9% (G1), 97.0% (G2; G3; G4) and 100.0% (G5; G6). Conclusions: MM adhesives (FuturabondVRU and AdheseVR Universal) showed similar and acceptable performance/success rates but also better clinical outputs than the SE-all-in-one adhesive (FuturabondVR DC), particularly in SE mode. Success and retention rates were similar and not dependent on materials or adhesion modes.Fundacão Fernando Pessoa, Porto, Portugalinfo:eu-repo/semantics/publishedVersio

Amplified and Homozygously Deleted Genes in Glioblastoma: Impact on Gene Expression Levels

This is an open-access article distributed under the terms of the Creative Commons Attribution License.-- et al.[Background]: Glioblastoma multiforme (GBM) displays multiple amplicons and homozygous deletions that involve relevant pathogenic genes and other genes whose role remains unknown. [Methodology]: Single-nucleotide polymorphism (SNP)-arrays were used to determine the frequency of recurrent amplicons and homozygous deletions in GBM (n = 46), and to evaluate the impact of copy number alterations (CNA) on mRNA levels of the genes involved. [Principal Findings]: Recurrent amplicons were detected for chromosomes 7 (50%), 12 (22%), 1 (11%), 4 (9%), 11 (4%), and 17 (4%), whereas homozygous deletions involved chromosomes 9p21 (52%) and 10q (22%). Most genes that displayed a high correlation between DNA CNA and mRNA levels were coded in the amplified chromosomes. For some amplicons the impact of DNA CNA on mRNA expression was restricted to a single gene (e.g., EGFR at 7p11.2), while for others it involved multiple genes (e.g., 11 and 5 genes at 12q14.1-q15 and 4q12, respectively). Despite homozygous del(9p21) and del(10q23.31) included multiple genes, association between these DNA CNA and RNA expression was restricted to the MTAP gene. [Conclusions]: Overall, our results showed a high frequency of amplicons and homozygous deletions in GBM with variable impact on the expression of the genes involved, and they contributed to the identification of other potentially relevant genes.This work was partially supported by Fundação para a Ciência e a Tecnologia, Portugal [FCT PIC/IC/83108/2007]; PhD fellowships from Fundação para a Ciência e a Tecnologia, Portugal [SFRH/BD/23086/2005, SFRH/BD/11820/2003]; Fundación Mutua Madrileña, Madrid, Spain [AP87692011]; and Instituto de Salud Carlos III, Ministerio de Ciencia e Innovación, Madrid, Spain [RTICC RD06/0020/0035].Peer Reviewe