Sustained gene expression in the retina by improved episomal vectors

Gene and cellular therapies are nowadays part of therapeutic strategies for the treatment of diverse pathologies. The drawbacks associated with gene therapy-low levels of transgene expression, vector loss during mitosis, and gene silencing-need to be addressed. The pEPI-1 and pEPito family of vectors was developed to overcome these limitations. It contains a scaffold/matrix attachment region, which anchors its replication to cell division in eukaryotic cells while in an extrachromosomal state and is less prone to silencing, due to a lower number of CpG motifs. Recent success showed that ocular gene therapy is an important tool for the treatment of several diseases, pending the overcome of the aforementioned limitations. To achieve sustained gene delivery in the retina, we evaluated several vectors based on pEPito and pEPI-1 for their ability to sustain transgene expression in retinal cells. These vectors stably transfected and replicated in retinal pigment epithelial (RPE) cells. Expression levels were promoter dependent with constitutive promoters cytomegalovirus immediate early promoter (CMV) and human CMV enhancer/human elongation factor 1 alpha promoter yielding the highest levels of transgene expression compared with the retina-specific RPE65 promoter. When injected in C57Bl6 mice, transgene expression was sustained for at least 32 days. Furthermore, the retina-specific RPE65 promoter showed higher efficiency in vivo compared to in vitro. In this study, we demonstrate that by combining tissue-specific promoters with a mitotic stable system, less susceptible to epigenetic silencing such as pEPito-based plasmids, we can achieve prolonged gene expression and a sustained therapeutic effect.Fundacao para a Ciencia e Tecnologia, Portugal [PEst/OE/EQB-LA 0023/2013, SFRH/BD/76873/2011, SFRH/BD/70318/2010, PTDC/SAU/BEB/098475/2008]; European Union [PIRG-GA-2009-249314

Biochemical composition and physicochemical properties of Moringa oleifera seed oil

Moringa oleifera tree has been recognized internationally for its nutritional, therapeutic and medicinal properties. Dry seeds are rich sources of oil with a high potential of commercial exploitation. The present study reports the physicochemical characterization, polyphenol content, DPPH radical scavenging capacity and fatty acid profile of moringa seed oil, and the chemical composition of the seed cultivated in Sonora, Mexico. Moisture, ash, protein and lipid contents in the seed were found to be 4.7, 5.8, 26 and 39%, respectively. The oil showed a refractive index of 1.4642. The saponification number was 183 mg KOH/g oil, iodine value: 75 g I/100 g of oil, acid value: 0.49 (% oleic acid). The polyphenol content was 0.137 mg of gallic acid equivalent/g and DPPH radical scavenging capacity was 87.39%. The moringa seed oil was rich (68%) in the major fatty acid, oleic acid (C18:1n9). Moringa oil extracted by sonication showed a fatty acid profile and physicochemical properties comparable to the oil from seeds grown in different regions of the world. The optimization of the oil extraction process on a large scale shows high potential, as the oil could be marketed as edible vegetable oil, for frying purposes, or as a functional ingredient

Draft Genome Sequence Of 11399, A Transformable Citrus-pathogenic Strain Of Xylella Fastidiosa

The draft genome of Xylella fastidiosa subsp. pauca strain 11399, a transformable citrus-pathogenic strain, is reported here. The 11399 genome size is 2,690,704 bp and has a G+C content of 52.7%. The draft genome of 11399 reveals the absence of four type I restriction-modification system genes. © 2016 Niza et al.4

The Atp-dependent Rna Helicase Hrpb Plays An Important Role In Motility And Biofilm Formation In Xanthomonas Citri Subsp. Citri

Background: RNA helicases are enzymes that catalyze the separation of double-stranded RNA (dsRNA) using the free energy of ATP binding and hydrolysis. DEAD/DEAH families participate in many different aspects of RNA metabolism, including RNA synthesis, RNA folding, RNA-RNA interactions, RNA localization and RNA degradation. Several important bacterial DEAD/DEAH-box RNA helicases have been extensively studied. In this study, we characterize the ATP-dependent RNA helicase encoded by the hrpB (XAC0293) gene using deletion and genetic complementation assays. We provide insights into the function of the hrpB gene in Xanthomonas citri subsp. citri by investigating the roles of hrpB in biofilm formation on abiotic surfaces and host leaves, cell motility, host virulence of the citrus canker bacterium and growth in planta. Results: The hrpB gene is highly conserved in the sequenced strains of Xanthomonas. Mutation of the hrpB gene (Δ;hrpB) resulted in a significant reduction in biofilms on abiotic surfaces and host leaves. Δ;hrpB also exhibited increased cell dispersion on solid medium plates. Δ;hrpB showed reduced adhesion on biotic and abiotic surfaces and delayed development in disease symptoms when sprayed on susceptible citrus leaves. Quantitative reverse transcription-PCR assays indicated that deletion of hrpB reduced the expression of four type IV pili genes. The transcriptional start site of fimA (XAC3241) was determined using rapid amplification of 5′-cDNA Ends (5′RACE). Based on the results of fimA mRNA structure predictions, the fimA 5′ UTR may contain three different loops. HrpB may be involved in alterations to the structure of fimA mRNA that promote the stability of fimA RNA. Conclusions: Our data show that hrpB is involved in adherence of Xanthomonas citri subsp. citri to different surfaces. In addition, to the best of our knowledge, this is the first time that a DEAH RNA helicase has been implicated in the regulation of type IV pili in Xanthomonas. © 2016 Granato et al.16

Soccer and homosexuality: the conflicts that lie within the affective game of the coach-adolescent athlete relationship

Discussions regarding homosexuality within a sports context are mediated by emotional nuances, especially in the world of soccer, where practitioners are required to have attributes such as strength, masculinity and aggressiveness. As a consequence, this work aimed to analyze the symbolic representation of homosexual relationships between adolescent soccer players and their coaches; and, furthermore, to understand the intention behind homoerotic relationships that are established between members of the management team and its players. In this study, researchers used the life history methodology, with participation from three adolescent athletes, ages 14, 15 and 16, and three former athletes, ages 26, 28 and 45, who we called speakers. Based on the collection and analysis of research participants’ testimonies, homosexual relations were found to have occurred between athletes and coaches, or between athletes and other members of the management team, as a part of a “social contract” that is exclusively linked to their professional objectives. These relationships appear to be purely objective; authentic affective links are absent, due apparently to the transient nature of the athletes in these groups. Too, this context is usually marked by the athlete’s physical and emotional vulnerability, and the agreements he makes within this context are accompanied with moral dilemmas, because such adolescents idealize the coach as a paternal archetype.“Futebol e homossexualidade: Os conflitos que se encontram dentro do jogo afetivo da relação treinador-atleta adolescente.” As discussões a respeito da homossexualidade no contexto esportivo são mediadas por nuanças emocionais, especialmente no mundo do futebol que exige de seus praticantes atributos de força, virilidade e agressividade. Por este motivo este trabalho teve como objetivos diagnosticar existência de relações homossexuais entre atletas adolescentes da modalidade de futebol e seus técnicos e sua representação simbólica e ainda, compreender a intencionalidade das relações homoafetivas que se estabelecem entre componentes da equipe diretiva e os jogadores. Esta pesquisa é de natureza qualitativa e a técnica adotada foi a “história de vida” e a participação de três atletas adolescentes em atividade, com idade de 14, 15 e 16 anos e três ex-atletas profissionais, com idades 26, 28 e 45 anos serviram de base metodológica. A partir da análise e tratamento dos depoimentos verificou-se que ocorreram relações homossexuais nas categorias de base envolvendo técnico e atleta, ou com outros membros da equipe diretiva, como parte dos contratos sociais, ligados exclusivamente a objetivos profissionais. Estas relações parecem ser puramente objetais, não parecendo existir vínculos afetivos duradouros, aparentemente, devido à natureza transitória de atletas nesses grupos. Também, nesses ambientes, muitas vezes marcados pela vulnerabilidade física e emocional dos atletas, os acertos para as relações homossexuais são permeadas por dilemas morais, tendo em vista que os atletas adolescentes idealizam o treinador como o arquétipo paterno ideal.“El fútbol y la homosexualidad: Los conflictos que se encuentran dentro del juego afectivo de la relación atleta-entrenador de los adolescentes.” Las discusiones sobre la homosexualidad en contexto deportivo están mediadas por matices emocionales, sobre todo en el mundo del fútbol que exige a sus jugadores atributos como la fuerza, la virilidad y los jugadores. En este estudio, los investigadores utilizaron la metodología de la historia de vida, con la participación de tres atletas adolescentes, con 14, 15 y 16 años, y tres ex atletas con 26, 28 y 45 años, que llamamos ‘oradores’. Basado en la recogida y análisis de testimonios de participantes en la investigación, se encontró que las relaciones homosexuales que se han producido entre los atletas y entrenadores, o entre atletas y otros miembros del equipo de gestión, son como parte de un “contrato social” que está vinculado exclusivamente a sus objetivos profesionales. Estas relaciones parecen ser solamente como un objeto; porque a veces no hay vínculos afectivos auténticos, debido aparentemente a la naturaleza transitoria de los atletas en estos grupos. Demasiado, este contexto es generalmente marcado por la vulnerabilidad física y emocional del deportista, y los acuerdos que hace dentro de este contexto se acompañan con dilemas morales, debido a que tales adolescentes idealizan el entrenador como un arquetipo paterno. la agresividad. Por lo tanto, este estudio tuvo como objetivo diagnosticar la existencia de relaciones homosexuales entre adolescentes atletas de fútbol y sus entrenadores y la representación simbólica de estas relaciones. También comprender la intencionalidad de las relaciones homoafetivas que se intercambian entre algunos componentes del equipo directivo y(undefined

Hard diffractive quarkonium hadroproduction at high energies

We present a study of heavy quarkonium production in hard diffractive process by the Pomeron exchange for Tevatron and LHC energies. The numerical results are computed using recent experimental determination of the diffractive parton density functions in Pomeron and are corrected by unitarity corrections through gap survival probability factor. We give predictions for single as well as central diffractive ratios. These processes are sensitive to the gluon content of the Pomeron at small Bjorken-x and may be particularly useful in studying the small-x physics. They may also be a good place to test the different available mechanisms for quarkonium production at hadron colliders.Comment: 7 pages, 3 figures, 1 table. Final version to be published in European Physical Journal

New triazine bridged triads based on BODIPY-porphyrin systems: extended absorption, efficient energy transfer and upconverted emission

Two novel triads connecting a BODIPY to ethylenediamine substituted porphyrins via triazine linker have been synthesized and characterized. One of the triads is a linear D-A structure with one BODIPY (D) and one porphyrin (A) bridged by the triazine linker and the other one is a branched A-D4 structure with the porphyrin core linked to four BODIPY units. The triads show extended absorption in the visible region with contributions from both porphyrin (Soret band centred at 410–430 nm) and BODIPY units (strong absorption at ≈ 502 nm) in good agreement with the expected molar ratio. Both triads exhibit linear and nonlinear optical properties featuring an efficient energy transfer from the BODIPY donor to the porphyrin acceptor. The nonlinear upconverted emission properties of the triads were studied by two-photon excitation in the Near-infrared (NIR, 710–930 nm). The maximum two-photon absorption cross-section values for the triads (40–70 GM) are larger than those typically reported in this wavelength range for porphyrins and BODIPY. Both the green emission of BODIPY (≈514 nm) and the red emission of porphyrins (650–750 nm) were observed under NIR excitation at 930 nm. The distinct features of triads, namely i) an extended absorption; ii) an efficient energy transfer and iii) the nonlinear upconverted emission featuring a large separation between the excitation and emission wavelengths could be beneficial for application in sensing and imaging procedures.publishe

Time-Fractional KdV Equation: Formulation and Solution using Variational Methods

In this work, the semi-inverse method has been used to derive the Lagrangian of the Korteweg-de Vries (KdV) equation. Then, the time operator of the Lagrangian of the KdV equation has been transformed into fractional domain in terms of the left-Riemann-Liouville fractional differential operator. The variational of the functional of this Lagrangian leads neatly to Euler-Lagrange equation. Via Agrawal's method, one can easily derive the time-fractional KdV equation from this Euler-Lagrange equation. Remarkably, the time-fractional term in the resulting KdV equation is obtained in Riesz fractional derivative in a direct manner. As a second step, the derived time-fractional KdV equation is solved using He's variational-iteration method. The calculations are carried out using initial condition depends on the nonlinear and dispersion coefficients of the KdV equation. We remark that more pronounced effects and deeper insight into the formation and properties of the resulting solitary wave by additionally considering the fractional order derivative beside the nonlinearity and dispersion terms.Comment: The paper has been rewritten, 12 pages, 3 figure