Bioprospecção de polissacarídeos sulfatados de macroalgas marinhas do litoral do Rio Grande do Norte: caracterização de uma heterofucana extraída da alga marron Sargassum filipendula que induz apoptose em células HeLa

The coast of Rio Grande do Norte has more than 100 species of seaweed, mostly unexplored regarding their pharmacological potential. The sulfated polysaccharides (PS) are by far the more seaweed compounds studied, these present a range of biological properties, such as anticoagulant activity, anti-inflammatory, antitumor and antioxidant properties. In this study, we extract sulfated polysaccharide rich-extracts of eleven algae from the coast of Rio Grande do Norte (Dictyota cervicornis; Dictiopterys delicatula; Dictyota menstruallis; Dictyota mertensis; Sargassum filipendula; Spatoglossum schröederi; Gracilaria caudata; Caulerpa cupresoides; Caulerpa prolifera; Caulerpa sertularioides e Codim isthmocladum), and these were evaluated for the potential anticoagulant, antioxidant and antiproliferative. All polysaccharide extracts showed activity for anticoagulant, antioxidant and/or antiproliferative activity, especially D. delicatula and S. filipendula, which showed the most prominent pharmacological potential, thereby being chosen to have their sulfated polysaccharides extracted. By fractionating method were obtained six fractions rich in sulfated polysaccharides to the algae D. delicatula (DD-0,5V, DD-0, 7V, DD-1,0v, DD-1,3v, DD-1,5v and DD-2,0) and five fractions to the alga S. filipendula (SF-0,5V, SF-0,7V, SF-1,0v, SF-1,5v and SF-2,0v). For the anticoagulant assay only the fractions of D. delicatula showed activity, with emphasis on DD-1, 5v that presented the most prominent activity, with APTT ratio similar to clexane® at 0.1 mg/mL. When evaluated the antioxidant potential, all fractions showed potential in all tests (total antioxidant capacity, hydroxyl and superoxide radicals scavenging, ferrous chelation and reducing power), however, the ability to chelate iron ions appears as the main mechanism antioxidant of sulfated polysaccharides from seaweed. In antiproliferative assay, all heterofucanas showed dose-dependent activity for the inhibition of cell proliferation of HeLa, however, with the exception of SF-0,7V, SF- 1,0v and SF-1,5v, all fractions showed antiproliferative activity against MC3T3, a normal cell line. The heterofucana SF-1,5V had its antiproliferative mechanism of action evaluated. This heterofucan induces apoptosis in HeLa cells by a pathway caspase independent, promoting the release of apoptosis Inducing Factor (AIF) in the cytosol, which in turn induces chromatin condensation and DNA fragmentation into 50Kb fragments. These results are significant in that they provide a mechanistic framework for further exploring the use of SF-1.5v as a novel chemotherapeutics against human cervical cancer.

The RGBarrier assay: the parallel study of gene regulatory element performance at defined chromosomal locations

Vertebrate genomes are organised into a variety of nuclear environments and chromatin states that have profound effects on the regulation of gene transcription. This variation presents a major challenge to the expression of transgenes for experimental research, genetic therapies and the production of biopharmaceuticals. The majority of transgenes succumb to transcriptional silencing by their chromosomal environment when they are randomly integrated into the genome, a phenomenon known as chromosomal position effect (CPE). It is not always feasible to target transgene integration to transcriptionally permissive “safe harbour” loci that favour transgene expression, so there remains an unmet need to identify gene regulatory elements that can be added to transgenes which protect them against CPE. Dominant regulatory elements (DREs) with chromatin barrier (or boundary) activity have been shown to protect transgenes from CPE. The HS4 element from the chicken beta-globin locus and the A2UCOE element from a human housekeeping gene locus have been shown to function as DRE barriers in a wide variety of cell types and species. Despite rapid advances in the profiling of transcription factor binding, chromatin states and chromosomal looping interactions, progress towards functionally validating the many candidate barrier elements in vertebrates has been very slow. This is largely due to the lack of a tractable and efficient assay for chromatin barrier activity. In this study, I have developed the RGBarrier assay system to test the chromatin barrier activity of candidate DREs at pre-defined isogenic loci in human cells. The RGBarrier assay consists in a Flp-based RMCE reaction for the integration of an expression construct, carrying candidate DREs, in a pre-characterised chromosomal location. The RGBarrier system involves the tracking of red, green and blue fluorescent proteins by flow cytometry to monitor on-target versus off-target integration and transgene expression. The analysis of the reporter (GFP) expression for several weeks gives a measure of the protective ability of each candidate elements from chromosomal silencing. This assay can be scaled up to test tens of new putative barrier elements in the same chromosomal context in parallel. The defined chromosomal contexts of the RGBarrier assays will allow for detailed mechanistic studies of chromosomal silencing and DRE barrier element action. Understanding these mechanisms will be of paramount importance for the design of specific solutions for overcoming chromosomal silencing in specific transgenic applications.

Projeto de uma ferramenta específica para cravação de terminais elétricos

A necessidade contínua de incrementar a produtividade em todos os sectores leva a que sejam utilizados equipamentos cada vez mais sofisticados, permitindo novas técnicas de abordagem dos processos de fabrico, velocidades mais elevadas e maior precisão no produto final. No entanto, quase todos os equipamentos necessitam de ferramentas adequadas, que tirem partido efetivamente das potencialidades dos equipamentos disponíveis. A Engenharia tem aqui um papel extremamente importante, já que terá de conceber as ferramentas atendendo à satisfação de um elevado número de requisitos, que passam genericamente por tirar o máximo partido dos fatores tecnológicos proporcionados pelos equipamentos, cumprir com a cadência de produção necessária, assegurar a qualidade estipulada, garantir a segurança dos colaboradores que com ela trabalham e permitir uma fácil montagem e afinação, reduzindo assim os tempos de preparação e as possibilidades de fabrico de peças fora das especificações. Este trabalho foi desenvolvido em torno de uma necessidade real, tendo sido estipulados os requisitos necessários pelo cliente e tendo sido elaborado todo o projeto da ferramenta em torno desses mesmos requisitos. Foi efetuada uma otimização da ferramenta ainda na fase de anteprojeto, permitindo minimizar o seu custo e maximizar o seu rendimento e aptidão para o fim em vista. Os materiais foram cuidadosamente selecionados, tendo em vista a utilização de cada componente e a relação custo‐benefício. Foi ainda efetuada uma orçamentação da mesma, assim como um plano de instruções para a operação e manutenção da ferramenta. Deste modo, este trabalho é o resumo de grande parte do conhecimento adquirido, quer ao longo da vida académica, quer ao longo da experiência profissional.

Draft Genome of Australian Environmental Strain WM 09.24 of the Opportunistic Human Pathogen Scedosporium aurantiacum

International audience

2005-2006 ACCOUNTABILITY REPORT

Each year South Carolina State University produces an annual accountability report for the South Carolina General Assembly and the Budget and Control Board. Included is an executive summary, agency discussion and analysis, and strategic planning documents.

Accountability Report

Each year South Carolina State University produces an annual accountability report for the South Carolina General Assembly and the Budget and Control Board. Included is an executive summary, agency discussion and analysis, and strategic planning documents.

2007-2008 ACCOUNTABILITY REPORT

Each year South Carolina State University produces an annual accountability report for the South Carolina General Assembly and the Budget and Control Board. Included is an executive summary, agency discussion and analysis, and strategic planning documents.

2008-2009 ACCOUNTABILITY REPORT

Each year South Carolina State University produces an annual accountability report for the South Carolina General Assembly and the Budget and Control Board. Included is an executive summary, agency discussion and analysis, and strategic planning documents.

2009-2010 ACCOUNTABILITY REPORT

Each year South Carolina State University produces an annual accountability report for the South Carolina General Assembly and the Budget and Control Board. Included is an executive summary, agency discussion and analysis, and strategic planning documents.

2010-2011 ACCOUNTABILITY REPORT

Each year South Carolina State University produces an annual accountability report for the South Carolina General Assembly and the Budget and Control Board. Included is an executive summary, agency discussion and analysis, and strategic planning documents.

2011-2012 ACCOUNTABILITY REPORT

Each year South Carolina State University produces an annual accountability report for the South Carolina General Assembly and the Budget and Control Board. Included is an executive summary, agency discussion and analysis, and strategic planning documents.

2012-2013 ACCOUNTABILITY REPORT

Each year South Carolina State University produces an annual accountability report for the South Carolina General Assembly and the Budget and Control Board. Included is an executive summary, agency discussion and analysis, and strategic planning documents.

Dual Quorum Quenching Capsules: Disrupting two bacterial communication pathways that lead to virulence

Healthcare Associated Infections (HAIs) in the United States, are estimated to cost nearly $10 billion annually. And, while device-related infections have decreased, the 60% attributed to pneumonia, gastrointestinal pathogens and surgical site infections (SSIs) remain prevalent. Furthermore, these are often complicated by antibacterial resistance that ultimately cause 2 million illnesses and 23,000 deaths in the US annually. Antibacterial resistance is an issue increasing in severity as existing antibiotics are losing effectiveness, and fewer new antibiotics are being developed. As a result, new methods of combating bacterial virulence are required. Modulating communications of bacteria can alter phenotype, such as biofilm formation and toxin production. Disrupting these communications provides a means of controlling virulence without directly interacting with the bacteria of interest, a strategy contrary to traditional antibiotics. Inter- and intra-species bacterial communication is commonly called quorum sensing because the communication molecules have been linked to phenotypic changes based on bacterial population dynamics. By disrupting the communication, a method called ‘quorum quenching’, bacterial phenotype can be altered. Virulence of bacteria is both population and species dependent; each species will secrete different toxic molecules, and total population will affect bacterial phenotype9. Here, the kinase LsrK and lactonase SsoPox were combined to simultaneously disrupt two different communication pathways with direct ties to virulence leading to SSIs, gastrointestinal infection and pneumonia. To deliver these enzymes for site-specific virulence prevention, two naturally occurring polymers were used, chitosan and alginate. Chitosan, from crustacean shells, and alginate, from seaweed, are frequently studied due to their biocompatibility, availability, self-assembly and biodegrading properties and have already been verified in vivo for wound-dressing. In this work, a novel functionalized capsule of quorum quenching enzymes and biocompatible polymers was constructed and demonstrated to have dual-quenching capability. This combination of immobilized enzymes has the potential for preventing biofilm formation and reducing bacterial toxicity in a wide variety of medical and non-medical applications.

Lander University Accountability Report

Each year Lander University produces an annual accountability report for the South Carolina General Assembly and the Budget and Control Board. Included is an executive summary, agency discussion and analysis, and strategic planning documents.

Lander University Accountability Report, 2005-2006

Each year Lander University produces an annual accountability report for the South Carolina General Assembly and the Budget and Control Board. Included is an executive summary, agency discussion and analysis, and strategic planning documents.