ABSTRACT
The eukaryotic green microalga Tetraselmis suecica is commonly used for aquaculture purposes because of its high stress tolerance and ease of culture in a wide spectrum of environments; they are therefore suitable candidates for biotechnology applications. To date, no data are available regarding chloroplast transformation vectors based on specific endogenous promoters and homologous targeting regions. We report on the identification of Tetraselmis suecica genes encoding the ribulose bisphosphate carboxylase/oxygenase large subunit protein, the photosystem II D1 protein and the ATP synthase CF1-beta subunit protein together with their untranslated regions (5'UTR, 3'UTR). The full-length ORFs of the putative genes with their regulatory sequences were obtained. We were also able to identify the downstream 3' end of the large subunit ribosomal RNA gene (23S) along with the 5S RNA end-to-end with the psbA gene on the complementary strand. The intergenic region between these genes appears to be a good target site for the integration of target proteins. Moreover, we identified a back-to-back promoter region among the rbcL and atpB genes. To assess the bidirectionality activities of both promoters, a dual reporter vector was constructed for Tetraselmis suecica transformation containing the cat and TurboGFP genes driven by the 5'rbcL/5'atpB divergent promoter. The vector included the 23S-5S and psbA nucleotide sequences as flanking regions. These flanking regions provided suitable insertion sites within the chloroplast genome for cassette integration via homologous recombination. Simultaneous expression of the chloramphenicol-resistant conferring gene and the gene coding for TurboGFP driven by 5'rbcL/5'atpB showed a potent natural bidirectional promoter as a reliable genetic tool.
Subject(s)
Chlorophyceae , Chloroplasts , Plasmids , Promoter Regions, Genetic , Ribulose-Bisphosphate Carboxylase/geneticsABSTRACT
El presente trabajo tiene como finalidad la valoración estudiantil respecto al uso de WhatsApp (WAP) y los Entornos Virtuales de Aprendizaje (EVA) en la Facultad de Odontología (FO), de la Udelar (Montevideo - Uruguay). Se realizó estudio descriptivo observacional en estudiantes de la generación 2016 - FO - UdelaR. Se utilizó una encuesta con 5 items en relación a tres aplicaciones tecnológicas (WhatsApp, Plataforma Moodle y Polimedias), fueron valorados de 0-12 puntos. En WhatsApp se consultó acerca de la posibilidad de consultar dudas, compartir imágenes, informarse, interactuar con el docente y compañeros, dando como resultado en cada item un puntaje promedio de 10.77-10.63-10.63-10.63 y 9.20 respectivamente. En EVA se consultaron los mismos ítems salvo el dos (fue acceder a presentaciones de clase), recibiendo un puntaje promedio de 8.53-11.27-10.33-9.47 y 5.53. Se percibe un alto grado de aceptación entre las diferentes tecnologías, con un mayor grado de aceptación en cuanto a la comunicación de WhatsApp frente a la plataforma Moodle.
The aim of this work was to have students evaluate the use of WhatsApp (WAP) and a Virtual Learning Environment (VLE) at the School of Dentistry, UdelaR (Montevideo - Uruguay). An observational descriptive study was conducted on students from the 2016 generation. A survey with five items was implemented, including three technological applications (WhatsApp, Moodle Platform and Polimedias), which were rated 0 to 12 points. As for WhatsApp, we asked about the possibility of clarifying doubts, sharing images, getting information, interacting with the teacher and classmates. The average score obtained was 10.77-10.63-10.63-10.63 and 9.20 for each item, respectively. In the VLE, the same items were included except for number two (access to class presentations), receiving an average score of 8.53-11.27-10.33-9.47 and 5.53. There is a high degree of acceptance of the different technologies, WhatsApp communication being more widely accepted than communication in the Moodle platform.
Subject(s)
Students, Dental , Education, DistanceABSTRACT
The angiosperm seed is composed of three genetically distinct tissues: the diploid embryo that originates from the fertilized egg cell, the triploid endosperm that is produced from the fertilized central cell, and the maternal sporophytic integuments that develop into the seed coat1. At the onset of embryo development in Arabidopsis thaliana, the zygote divides asymmetrically, producing a small apical embryonic cell and a larger basal cell that connects the embryo to the maternal tissue2. The coordinated and synchronous development of the embryo and the surrounding integuments, and the alignment of their growth axes, suggest communication between maternal tissues and the embryo. In contrast to animals, however, where a network of maternal factors that direct embryo patterning have been identified3,4, only a few maternal mutations have been described to affect embryo development in plants5-7. Early embryo patterning in Arabidopsis requires accumulation of the phytohormone auxin in the apical cell by directed transport from the suspensor8-10. However, the origin of this auxin has remained obscure. Here we investigate the source of auxin for early embryogenesis and provide evidence that the mother plant coordinates seed development by supplying auxin to the early embryo from the integuments of the ovule. We show that auxin response increases in ovules after fertilization, due to upregulated auxin biosynthesis in the integuments, and this maternally produced auxin is required for correct embryo development.
Subject(s)
Arabidopsis/growth & development , Indoleacetic Acids/pharmacology , Plant Development/drug effects , Seeds/drug effects , Arabidopsis/metabolism , Seeds/growth & developmentABSTRACT
With the purpose of expanding the structural variety of chemical compounds available as pharmacological tools for the treatment of Alzheimer's disease, we synthesized and evaluated a novel series of indole-benzoxazinones (Family I) and benzoxazine-arylpiperazine derivatives (Family II) for potential human acetylcholinesterase (hAChE) inhibitory properties. The most active compounds 7a and 7d demonstrated effective inhibitory profiles with Ki values of 20.3 ± 0.9 µM and 20.2 ± 0.9 µM, respectively. Kinetic inhibition assays showed non-competitive inhibition of AChE by the tested compounds. According to our docking studies, the most active compounds from both series (Families I and II) showed a binding mode similar to donepezil and interact with the same residues.
Subject(s)
Acetylcholinesterase/drug effects , Benzoxazines/pharmacology , Cholinesterase Inhibitors/pharmacology , Piperazines/pharmacology , Acetylcholinesterase/metabolism , Alzheimer Disease/drug therapy , Benzoxazines/chemical synthesis , Benzoxazines/chemistry , Cholinesterase Inhibitors/chemical synthesis , Cholinesterase Inhibitors/chemistry , Donepezil , Drug Design , Humans , Indans/pharmacology , Molecular Docking Simulation , Piperazines/chemical synthesis , Piperazines/chemistry , Piperidines/pharmacology , Protein Binding , Structure-Activity RelationshipABSTRACT
Diversas especies de abejas sin aguijón producen miel de botija en Venezuela, la cual no está incluida en las normas venezolanas de calidad. Una de estas especies, la Meli po na favosa, vive en la Península de Paraguaná, donde se conoce como erica o maba. A fin de conocer las mieles de M. favosa, se realizó la determinación de su origen botánico, su caracterización físicoquímica según los métodos de la norma COVENIN 2136-84 para miel de abejas y contenido de nitrógeno por microkjeldahl, y de actividad biológica con métodos espectrofotométricos y de concentración inhibitoria mínima, en seis mieles. Son mieles claras de color ámbar entre 18 y 79 mm Pfund. La composición físicoquímica varió así: acidez libre 12,72-95,86 meq/kg, pH 3,53-4,44, humedad 25,40-32,00 g agua/100 g, 0,01-0.16 g ce ni zas/100g, 10,48-57,55 mgN/100 g, 62,60-69,50 g azúcares reductores/100 g, 0,60-5,10 g sacarosa aparente/100 g, flavonoides 0,10-8,15 mgEQ/100g miel, y polifenoles 51,50-217,19 mgEAG/100 g. La actividad antibac teriana (g miel/100 mL medio de cultivo) fue mayor contra E. coli (12.50-50,00) que contra S. aureus (50,00- 50,00). La actividad antioxidante se ubicó en un rango bajo a alto para mieles, con 45,91-227,92 mmoles equivalentes de Trolox/100 g. El análisis melisopalinológico indicó polen dominante del género Portulaca y la especie Carica papaya.
Diverse species of stingless bees produce por honey in Venezuela,which is not included in the Venezuelan regulations of honey quality. One of these species , the Melipona favosa, lives in the Paraguaná Peninsula, where it is known as erica or maba. In order to know the honeys of M. favosa, the botanical origin was determined, their physicochemical characterization was done according to the methods of the norm COVENIN 2136-84 for honey, the nitrogen content by microkjeldahl, and biological activity by spectrophotometric and minimum inhibitory concentration methods, in six ho neys. They are light honey amber color between 18 and 79 mm Pfund. The physicochemical composition varied as follows: free acidity 12.72- 95.86 meq/kg, pH 3.53-4.44, moisture 25.40-32.00 g water/100 g, 0.01-0.16 g ash/100g, 10.48-57.55 mgN/100 g, 62.60-69.50 g reducing su gars/100 g, 0.60-5.10 g apparent sucrose/100 g, flavonoids 0.10-8.15 mgQE/100g honey, and polyphenols 51.50-217.19 mgGAE/100 g. The antibacterial ac tivity (g ho ney/100 mLculture media) was higher against E. coli (12.50-50.00) than S. aureus (50.00-50.00). The antioxidant activity was in a low to high range for honeys, with 45.91- 227.92 mmoles Trolox equivalents/100 g.
Subject(s)
Humans , Male , Female , Anti-Bacterial Agents/therapeutic use , Antioxidants/administration & dosage , Bees/classification , Public Health , HoneyABSTRACT
At present, there is strong commercial demand for recombinant proteins, such as antigens, antibodies, biopharmaceuticals, and industrial enzymes, which cannot be fulfilled by existing procedures. Thus, an intensive search for alternative models that may provide efficiency, safety, and quality control is being undertaken by a number of laboratories around the world. The chloroplast of the eukaryotic microalgae Haematococcus pluvialis Flotow has arisen as a candidate for a novel expression platform for recombinant protein production. However, there are important drawbacks that need to be resolved before it can become such a system. The most significant of these are chloroplast genome characterizations, and the development of chloroplast transformation vectors based upon specific endogenous promoters and on homologous targeting regions. In this study, we report the identification and characterization of endogenous chloroplast sequences for use as genetic tools for the construction of H. pluvialis specific expression vectors to efficiently transform the chloroplast of this microalga via microprojectile bombardment. As a consequence, H. pluvialis shows promise as a platform for expressing recombinant proteins for biotechnological applications, for instance, the development of oral vaccines for aquaculture.
ABSTRACT
La normativa nacional e internacional permite adicionar a las mezclas deshidratadas para caldos y sopas resaltadores del sabor tales como glutamato monosódico (GMS) y los ácidos inosínico y guanílico o sus sales fosfatadas (IMF y GMF, respectivamente), solos o combinados para lograr un efecto sinergístico. Los objetivos de este estudio fueron: 1) determinar mediante un panel sensorial, el efecto sinergístico en el sabor de una sopa de pollo deshidratada cuando se usa mezcla de resaltadores y 2) cuantificar los 5-ribonucleótidos en dicha matriz. La intensidad del sabor a pollo se determinó empleando un panel de 6 personas previamente entrenadas. Los 5-ribonucleótidos se cuantificaron empleando HPLC. Los resultados con el panel entrenado comprobaron que la combinación utilizada de GMS, IMF y GMF potencia significativamente (p<0,05) el sabor de la sopa de pollo deshidratada. El análisis químico de los 5´-ribonucleótidos reflejó un porcentaje de recuperación de 93,6 por ciento para GMF y 90,5 por ciento para IMF
