EARLY FLOWERING 3 interactions with PHYTOCHROME B and PHOTOPERIOD1 are critical for the photoperiodic regulation of wheat heading time.

The photoperiodic response is critical for plants to adjust their reproductive phase to the most favorable season. Wheat heads earlier under long days (LD) than under short days (SD) and this difference is mainly regulated by the PHOTOPERIOD1 (PPD1) gene. Tetraploid wheat plants carrying the Ppd-A1a allele with a large deletion in the promoter head earlier under SD than plants carrying the wildtype Ppd-A1b allele with an intact promoter. Phytochromes PHYB and PHYC are necessary for the light activation of PPD1, and mutations in either of these genes result in the downregulation of PPD1 and very late heading time. We show here that both effects are reverted when the phyB mutant is combined with loss-of-function mutations in EARLY FLOWERING 3 (ELF3), a component of the Evening Complex (EC) in the circadian clock. We also show that the wheat ELF3 protein interacts with PHYB and PHYC, is rapidly modified by light, and binds to the PPD1 promoter in planta (likely as part of the EC). Deletion of the ELF3 binding region in the Ppd-A1a promoter results in PPD1 upregulation at dawn, similar to PPD1 alleles with intact promoters in the elf3 mutant background. The upregulation of PPD1 is correlated with the upregulation of the florigen gene FLOWERING LOCUS T1 (FT1) and early heading time. Loss-of-function mutations in PPD1 result in the downregulation of FT1 and delayed heading, even when combined with the elf3 mutation. Taken together, these results indicate that ELF3 operates downstream of PHYB as a direct transcriptional repressor of PPD1, and that this repression is relaxed both by light and by the deletion of the ELF3 binding region in the Ppd-A1a promoter. In summary, the regulation of the light mediated activation of PPD1 by ELF3 is critical for the photoperiodic regulation of wheat heading time.

Dynamics of Polyphenol Biosynthesis by Calli Cultures, Suspension Cultures and Wild Specimens of the Medicinal Plant Ligaria cuneifolia (Ruiz & Pav.) Tiegh. (Loranthaceae). Analysis of Their Biological Activity.

Ligaria cuneifolia (R. et P.) Tiegh. (Loranthaceae) is a South American hemiparasitic species with antioxidant, antitumoral, antimicrobial, and antilipidemic activities attributed to its polyphenolic content. We studied the polyphenolic pattern of L. cuneifolia during different phenological stages: flowering, fruiting, and post-fruiting. The highest total phenolic content was found in stems at post-fruiting (214 ± 12.1 mg gallic acid eq·g-1 DW) and fruiting (209 ± 13.7 mg gallic acid eq·g-1 DW), followed by post-fruiting leaves (207 ± 17.5 mg gallic acid eq·g-1 DW). Flavonoids accumulated at higher levels in leaves and hydroxycinnamic acids in leaves at flowering and post-fruiting. The polyphenolic pattern was similar between organs from wild plants and in vitro cultures, although at a significantly lower level in the latter ones. The performance of calli growing under a 16 h photoperiod in a modified White medium with 1-naphthalene acetic acid (2.50 µM) and Kinetin (9.20 µM) was better than in the dark. When calli grew in media only with auxins (IAA, NAA, and 2,4-D, all at 2.50 µM concentration), its growth and polyphenolic content improved. Cell suspensions with 2.50 µM NAA and 9.20 µM KIN grew slowly and produced very small amounts of polyphenols. As for the antioxidant activity, it was detected in all samples (approximately 1000 µmol trolox eq·g-1 DW) except fruits, where a lower value was found (328 µmol trolox eq·g-1 DW). In vitro cultures have the lowest antioxidant activity when compared to methanolic extracts from organs of wild specimens. Finally, antimutagenic or mutagenic activity in wild plants and in vitro culture extracts was not detected by the Ames test.

Adulto mayor: envejecimiento, discapacidad, cuidado y centros día. Revisión de tema / Older Adult: Aging, Disability, Care, and Day Centers. A Review

RESUMEN Objetivo: Describir el proceso y modelos de envejecimiento, discapacidad, cuidado y centros día para atención de población adulta mayor. Metodología: Revisión narrativa de tema utilizando bases de datos como PubMed, información de la Organización Mundial de la Salud, guías de geriatría, revistas, artículos científicos y trabajos de grado acerca de los temas de vejez, aumento de la población y centros día. Resultados: El envejecimiento como proceso natural hace parte del ciclo de vida y comprende un amplio conjunto de procesos biológicos, psicológicos y sociales. Los centros día son una opción para no institucionalizar al adulto mayor dependiente, y en el adulto mayor sano, una opción de actividad, recreación y dignificación. Además, tienen objetivos dirigidos al cuidador. Actualmente el envejecimiento poblacional es una realidad mundial en la que los centros día pueden ser parte de la respuesta a la hora de cubrir las necesidades de cuidado, dignificación e integración de la población adulta mayor. Conclusión: Una opción para brindar cuidado integral de los adultos mayores son los centros día, los cuales son una alternativa intermedia entre conservar su ambiente habitual/ familiar y casos de institucionalización. El tema de los centros día para adultos mayores es un tópico que continúa en desarrollo, respecto a los cuales existen varias definiciones, formas de trabajo y procesos terapéuticos que se pueden llevar a cabo en estos. Es necesario conservar y fortalecer la salud y bienestar de los adultos mayores, y promover procesos de envejecimiento exitoso, saludable y/o activo.

ABSTRACT Objective: Describe the process and models of aging, disability, care and day-care centers for the elderly population. Methodology: A narrative review of the topic using databases such as PubMed, information from the World Health Organization, geriatric guides, journals, scientific articles, and graduate projects on the issues of old age, population growth, and daycare centers. Results: Aging as a natural process is part of the life cycle and includes a broad set of biological, psychological, and social processes. Daycare centers are an option not to institutionalize the dependent older adult, and, for the healthy older adult, an option for activity, recreation, and dignity. In addition, they have goals directed at the caregiver. Currently, population aging is a global reality where daycare centers can be part of the answer to meeting the needs of care, dignity, and integration of older adults. Conclusion: Care goes beyond medical care. One option to provide comprehensive care for older adults is daycare centers, which are an intermediate alternative between preserving their usual/family environment and cases of institutionalization. The topic of daycare centers for older adults is a topic that continues to develop. There are various definitions, ways of working, and therapeutic processes that can be carried out. It is necessary to preserve and strengthen the health and well-being of older adults, and to promote successful, healthy, and active aging processes.

Characterization of the Bacteriophage vB_VorS-PVo5 Infection on Vibrio ordalii: A Model for Phage-Bacteria Adsorption in Aquatic Environments.

A mathematical first-order difference equation was designed to predict the dynamics of the phage-bacterium adsorption process in aquatic environments, under laboratory conditions. Our model requires knowledge of bacteria and bacteriophage concentrations and the measurements of bacterial size and velocity to predict both the number of bacteriophages adsorbed onto their bacterial host and the number of infected bacteria in a given specific time. It does not require data from previously performed adhesion experiments. The predictions generated by our model were validated in laboratory. Our model was initially conceived as an estimator for the effectiveness of the inoculation of phages as antibacterial therapy for aquaculture, is also suitable for a wide range of potential applications.

Los sistemas vegetales como plataformas de producción de proteínas de interés para la industria farmacéutica / Plant platforms for producing recombinant proteins for the pharmaceutical industry

La biotecnología vegetal logró armonizar los nuevos desarrollos relacionados con los sistemas productivos vegetales con la necesidad de proteínas recombinantes de la industria farmacéutica. La plataforma vegetal resultó una alternativa válida de producción principalmente por poseer la maquinaria adecuada para la síntesis proteica, incluyendo glicoproteínas y proteínas multiméricas. Dentro de las ventajas de estos sistemas se encuentran: la bioseguridad, pues no hay posibilidad de contaminación con patógenos, priones, oncogenes o endotoxinas; la facilidad de escalado, en el caso de los cultivos a campo, pues se realiza con bajos incrementos en costos; el desarrollo del proceso productivo en condiciones ambientales controladas, en el caso de los cultivos in vitro, pudiendo trabajarse en condiciones de Buenas Prácticas de Laboratorio y Buenas Prácticas de Manufactura y en el caso de la expresión transitoria, la rápida producción de proteínas terapéuticas. Una de las limitaciones de las proteínas producidas en vegetales es la diferencia en cuanto a la glicosilación respecto a las de origen animal. Esto ha logrado solucionarse mediante la inactivación de glicosiltransferasas específicas de plantas o su complementación con glicosiltransferasas animales heterólogas. Otra de las limitaciones para la explotación comercial de la producción de proteínas en plantas son sus bajos rendimientos. No obstante, se han desarrollado estrategias a nivel genético, del mecanismo de expresión (estable o transitoria), de las condiciones y del sistema de cultivo para incrementarlos. Así es como mediante transformación estable o transitoria de diferentes especies se ha logrado producir una amplia gama de proteínas recombinantes en cultivos agronómicos o cultivos in vitro. Proteínas funcionales de origen animal tales como anticuerpos, antígenos vacunales, citoquinas, hormonas de crecimiento, enzimas, biopolímeros y otras proteínas industriales han sido expresadas en especies tan diversas como Nicotiana tabacum, N. benthamiana, Daucus carota, Lactuca sativa, Lycopersicon esculentum, Solanum tuberosum, Oryza sativa, Zea mays, Glycine max, entre otras.

Plant biotechnology succeeded in combining new developments related to plant production systems with the need for recombinant proteins in the pharmaceutical industry. The plant platform turned out to be a valid production alternative mainly because it had adequate machinery for protein synthesis, including glycoproteins and multimeric proteins. Among the advantages of these systems are: biosafety, since there is no possibility of contamination with pathogens, prions, oncogenes or endotoxins; the ease of scaling up, in the case of field crops, as it is done with low cost increases; the development of the production process under controlled environmental conditions, in the case of in vitro cultures, being able to work under conditions of Good Laboratory Practices and Good Manufacturing Practices and the transitory expression, in the case of production of therapeutic proteins, which is done very quickly. One of the limitations of proteins produced in vegetables is the difference in glycosylation with respect to those of animal origin. This has been solved by the inactivation of plant-specific glycosyltransferases and/or their complementation with heterologous animal glycosyltransferases. Another limitation of the commercial exploitation of protein production in plants is its low yields. However, strategies have been developed at the genetic level, the expression mechanism (stable or transient), the culture conditions, and the culture system to increase them. This is how, through a stable or transient transformation of different species, a wide range of recombinant proteins has been produced in agronomic or in vitro cultures. Functional proteins of animal origin such as antibodies, antigens, cytokines, growth hormones, enzymes, biopolymers, and other industrial proteins have been expressed in species as diverse as Nicotiana tabacum, N. benthamiana, Daucus carota, Lactuca sativa, Lycopresicon esculentum, Solanum tuberosum, Oryza sativa, Zea mays, Glycine max, among others.

Expression of an scFv antibody fragment in Nicotiana benthamiana and in vitro assessment of its neutralizing potential against the snake venom metalloproteinase BaP1 from Bothrops asper.

Human accidents with venomous snakes represent an overwhelming public health problem, mainly in rural populations of underdeveloped countries. Their high incidence and the severity of the accidents result in 81,000 to 138,000 deaths per year. The treatment is based on the administration of purified antibodies, produced by hyper immunization of animals to generate immunoglobulins (Igs), and then obtained by fractionating hyper immune plasma. The use of recombinant antibodies is an alternative to conventional treatment of snakebite envenoming, particularly the Fv fragment, named the single-chain variable fragment (scFv). We have produced recombinant single chain variable fragment scFv against the venom of the pit viper Bothrops asper at high levels expressed transiently and stably in transgenic plants and in vitro cultures that is reactive to BaP1 (a metalloproteinase from B. asper venom). The yield from stably transformed plants was significantly (p > 0.05) higher than the results in from transient expression. In addition, scFvBaP1 yields from systems derived from stable transformation were: transgenic callus 62 µg/g (±2); biomass from cell suspension cultures 83 µg/g (±0.2); culture medium from suspensions 71.75 mg/L (±6.18). The activity of scFvBaP1 was confirmed by binding and neutralization of the fibrin degradation induced by BnP1 toxins from B. neuwiedi and by Atroxlysin Ia from B. atrox venoms. In the present work, we demonstrated the potential use of plant cells to produce scFvBaP1 to be used in the future as a biotechnological alternative to horse immunization protocols to produce anti-venoms to be used in human therapy against snakebites.

Expression of an scFv antibody fragment in Nicotiana benthamiana and in vitro assessment of its neutralizing potential against the snake venom metalloproteinase BaP1 from Bothrops asper

Human accidents with venomous snakes represent an overwhelming public health problem, mainly in ruralpopulations of underdeveloped countries. Their high incidence and the severity of the accidents result in 81,000to 138,000 deaths per year. The treatment is based on the administration of purified antibodies, produced byhyper immunization of animals to generate immunoglobulins (Igs), and then obtained by fractionating hyperimmune plasma. The use of recombinant antibodies is an alternative to conventional treatment of snakebiteenvenoming, particularly the Fv fragment, named the single-chain variable fragment (scFv). We have producedrecombinant single chain variable fragment scFv against the venom of the pit viperBothrops asperat high levelsexpressed transiently and stably in transgenic plants andin vitrocultures that is reactive to BaP1 (a metallo-proteinase fromB. aspervenom). The yield from stably transformed plants was significantly (p > 0.05) higherthan the results in from transient expression. In addition, scFvBaP1 yields from systems derived from stabletransformation were: transgenic callus 62µg/g ( ± 2); biomass from cell suspension cultures 83µg/g ( ± 0.2);culture medium from suspensions 71.75 mg/L ( ± 6.18). The activity of scFvBaP1 was confirmed by binding andneutralization of thefibrin degradation induced by BnP1 toxins fromB. neuwiediand by Atroxlysin Ia fromB.atroxvenoms. In the present work, we demonstrated the potential use of plant cells to produce scFvBaP1 to beused in the future as a biotechnological alternative to horse immunization protocols to produce anti-venoms tobe used in human therapy against snakebites.

Expression of an scFv antibody fragment in Nicotiana benthamiana and in vitro assessment of its neutralizing potential against the snake venom metalloproteinase BaP1 from Bothrops asper

Human accidents with venomous snakes represent an overwhelming public health problem, mainly in ruralpopulations of underdeveloped countries. Their high incidence and the severity of the accidents result in 81,000to 138,000 deaths per year. The treatment is based on the administration of purified antibodies, produced byhyper immunization of animals to generate immunoglobulins (Igs), and then obtained by fractionating hyperimmune plasma. The use of recombinant antibodies is an alternative to conventional treatment of snakebiteenvenoming, particularly the Fv fragment, named the single-chain variable fragment (scFv). We have producedrecombinant single chain variable fragment scFv against the venom of the pit viperBothrops asperat high levelsexpressed transiently and stably in transgenic plants andin vitrocultures that is reactive to BaP1 (a metallo-proteinase fromB. aspervenom). The yield from stably transformed plants was significantly (p > 0.05) higherthan the results in from transient expression. In addition, scFvBaP1 yields from systems derived from stabletransformation were: transgenic callus 62µg/g ( ± 2); biomass from cell suspension cultures 83µg/g ( ± 0.2);culture medium from suspensions 71.75 mg/L ( ± 6.18). The activity of scFvBaP1 was confirmed by binding andneutralization of thefibrin degradation induced by BnP1 toxins fromB. neuwiediand by Atroxlysin Ia fromB.atroxvenoms. In the present work, we demonstrated the potential use of plant cells to produce scFvBaP1 to beused in the future as a biotechnological alternative to horse immunization protocols to produce anti-venoms tobe used in human therapy against snakebites.

Laryngeal chondrosarcoma, case report and literature review.

INTRODUCTION: Chondrosarcoma constitutes 0.2% of all malignant tumors of the larynx. Many surgeons treat it with total laryngectomy due to the limited experience with this neoplasm because its rarity, and although several conservative approaches have been proposed, the data of these techniques are limited and based on retrospective series. PRESENTATION OF CASE: A 52-year-old male with a transglottic submucosal tumor and glottic stenosis in fiberoptic examination showed by tomography a laryngeal tumor that infiltrates vocal cords, glottis, cricoid and thyroid cartilage of 3 × 2.7 × 4 cm. Patient was submitted to total laryngectomy with selective bilateral neck dissection because obstructive tumor. Pathology reported a cricoid cartilage tumor consistent with grade 2 chondrosarcoma. DISCUSSION: Biopsy by laryngoscopy is considered the standard procedure for the diagnosis of laryngeal tumors, however the need for general anesthesia and the difficulty in intubation in some patients with large tumors make difficult to obtain an adequate biopsy in some cases with submucosal tumor. Conservative surgeries should be individualized based on the size and location of the tumor as well as on the patient's general conditions. Radical treatment is recommended for high-grade and large tumors in which conservative surgery would destabilize the cricoid ring. CONCLUSION: There is no diagnostic and treatment approach established for laryngeal chondrosarcoma, we believe that percutaneous biopsy would be the diagnostic test of choice because it is less invasive and has a high sensitivity and specificity; it could also identify patients who are candidates for conservative surgeries.

Influencia de reguladores de crecimiento sobre el establecimiento de cultivos in vitro de Salvia hispanica L. ("chía") y sobre su contenido de ácidos grasos / Influence of growth regulators on the establishment of in vitro cultures of Salvia hispanica L. ("chia") and its content of fatty acids

Salvia hispanica L. (Labiateae), comúnmente conocida como "chía", es una especie herbácea anual cuyo cultivo está ampliamente extendido por América del Sur. Es utilizada como cultivo industrial y para la ela-boración de alimentos funcionales por su contenido en proteínas, antioxidantes, fibras y lípidos esenciales. El aceite de sus semillas contiene la mayor proporción (68 %) de omega-3 que cualquier fuente vegetal conocida. El objetivo de este trabajo fue establecer cultivos in vitro de "chía" y analizar la influencia de diferentes reguladores de crecimiento sobre la inducción de callos, sobre el crecimiento de estos y sobre su contenido de ácidos grasos. Se iniciaron cultivos de callos in vitro a partir de explantos de tallos sin nudos y de hojas de plántulas axénicas de 20 días de edad, utilizando 3 tratamientos de reguladores de crecimiento diferentes. El medio Murashige & Skoog modificado (MSRT) con el agregado de ácido 2,4-diclorofenoxia-cético (2,4-D) a una concentración de 2,25 µM y un fotoperíodo de 16 horas fueron las condiciones óptimas para la inducción de callos. Para el mantenimiento de los callos el tratamiento más adecuado resultó ser bencilaminopurina (BAP) a una concentración de 1 µM. La cinética de crecimiento se caracterizó por un período de latencia hasta el día 20 de cultivo, seguido de un período de crecimiento exponencial entre los días 20 y 54. El tratamiento con 2,4-D (2,25 µM) mostró la más alta velocidad específica de crecimiento (0,22 ± 0,01 /día), el tiempo de duplicación más bajo (31,51 ± 1,00 día) y la mayor biomasa máxima (1,46 ± 0,01 g PF). El contenido de ácidos grasos en los callos de "chía" fue 0,83 % en promedio de todos los tratamientos después de 6 meses en cultivo y no mostró variaciones significativas (0,2 % - 0,3 %) entre los tratamientos aplicados (p < 0,05). (AU)

Kemp Eliminase Activity of Ketosteroid Isomerase.

Kemp eliminases represent the most successful class of computationally designed enzymes, with rate accelerations of up to 109-fold relative to the rate of the same reaction in aqueous solution. Nevertheless, several other systems such as micelles, catalytic antibodies, and cavitands are known to accelerate the Kemp elimination by several orders of magnitude. We found that the naturally occurring enzyme ketosteroid isomerase (KSI) also catalyzes the Kemp elimination. Surprisingly, mutations of D38, the residue that acts as a general base for its natural substrate, produced variants that catalyze the Kemp elimination up to 7000-fold better than wild-type KSI does, and some of these variants accelerate the Kemp elimination more than the computationally designed Kemp eliminases. Analysis of the D38N general base KSI variant suggests that a different active site carboxylate residue, D99, performs the proton abstraction. Docking simulations and analysis of inhibition by active site binders suggest that the Kemp elimination takes place in the active site of KSI and that KSI uses the same catalytic strategies of the computationally designed enzymes. In agreement with prior observations, our results strengthen the conclusion that significant rate accelerations of the Kemp elimination can be achieved with very few, nonspecific interactions with the substrate if a suitable catalytic base is present in a hydrophobic environment. Computational design can fulfill these requirements, and the design of more complex and precise environments represents the next level of challenges for protein design.

The expression of the 14D9 catalytic antibody in suspended cells of Nicotiana tabacum cultures increased by the addition of protein stabilizers and by transference from Erlenmeyer flasks to a 2-L bioreactor.

The effect of two protein stabilizers (polyvinylpyrrolidone [PVP] and gelatine) on growth and 14D9 yield of Nicotiana tabacum cell suspension cultures (Ab-KDEL and sec-Ab) was analyzed. The addition of PVP at a concentration of 1.0 g L(-1) produced the highest total 14D9 yield (biomass + culture medium) in the Ab-KDEL line (4.82% total soluble protein [TSP]). With the addition of gelatine, the highest total 14D9 yield (2.48% TSP) was attained in the Ab-KDEL line at 5.0 g L(-1) gelatine. When the Ab-KDEL suspended cells were cultured in a 2-L bioreactor, the highest 14D9 yield was 8.1% TSP at a 5% w/v inoculum size, which was the best 14D9 yield so far obtained in the platforms tested (E. coli, N. tabacum leaves and seeds, N. tabacum hairy roots, and cell suspension cultures).

La comprensión verbal y la lectura en niños con y sin retraso lector / Verbal Comprehension and Reading in Children with Reading Delay / A compreensÃo verbal e a leitura em crianças com e sem retraso leitor

Objetivo. Describir diferencias entre el desempeño en tareas de lectura y de comprensión verbal en niños con y sin retraso lector, así como la relación entre ambos tipos de desempeños. Método. Se utilizaron tareas de lectura de la Evaluación neuropsicológica infantil (ENI) y el índice de Comprensión Verbal (ICV) de la escala de inteligencia Wechsler (WISC-IV) para niños. Los participantes fueron 40 niños de ambos sexos, entre 7 y 9 años de edad, quienes cursaban entre 2° y 4° grado de primaria en un colegio de estrato socioeconómico bajo de la ciudad de Cali, con un coeficiente intelectual (CI) promedio de 86. Los participantes fueron divididos en dos grupos, 20 niños que presentaban retraso lector y 20 niños con un desarrollo lector típico, según los puntajes obtenidos en las tareas de lectura de la ENI. Resultados. Se encontraron diferencias en el rendimiento de ambos grupos de niños evaluados, así como una correlación significativa entre el ICV y el subdominio de comprensión de la ENI. Conclusión. Estos resultados son acordes con la hipótesis de que el desarrollo del lenguaje oral es uno de los principales facilitadores en la adquisición del aprendizaje de la lectura, puesto que en la medida en que el niño comprende la semántica oral, estará con mejor preparación para dotar de significado lo leído.

Objective. This study aims to describe differences between performance in reading tasks and the level of verbal comprehension in subjects, with and without reading delay, as the relationship between both measures. Method. For this purpose, the reading subtests of the Evaluación Neuropsicológica Infantil (ENI) and the verbal comprehension index of the Wechsler Intelligence Scale for Children (WISC-IV) were used. The sample consisted of 40 children, aged from 7 to 9 years old, in 2nd to 4th grade, from a low socioeconomic strata school in Cali, and with an intelligence quotient (IQ) which averaged 86 Participants were divided into two groups; 20 with reading disabilities and 20 who were normal readers, according to the scores obtained in the ENI reading tasks. Results. The results showed differences between the perfomances of both groups of children, as well as a significant positive correlation between ICV and reading comprehension of the ENI. Conclusion. These results are in accordance with the hypothesis that states that oral language development is one of the most important facilitators of learning in reading acquisition, because while children understand what they say and what they are told, they will be able to make sense of what they read.

Escopo. Descrever as diferenças entre o desempenho em tarefas de leitura e compreensÃo verbal em crianças com e sem atraso, ea relaçÃo entre os dois tipos de performances. Metodologia. Foram utilizadas tarefas de leitura da AvaliaçÃo Neuropsicológica Infantil (ENI por suas siglas em espanhol) e o índice de CompreensÃo Verbal (ICV) da escada de inteligência Wechsler (WISC-IV) para crianças. Os participantes foram 40 crianças de ambos sexos, entre 7 e 9 anos de idade, que cursavam entre 2 e 4 grado de primária em um colégio de baixa extraçÃo socioeconómica na cidade de Cali, com um coeficiente intelectual (CI) com 86 como média. Os participantes foram divididos em dois grupos, 20 crianças que presentavam retraso leitor e 20 crianças com um desenvolvimento leitor típico, segundo as pontuações obtidas nas tarefas de leitura da ENI. Resultados. em grupos Mabos, ICV e compreensÃo do IPD foram encontrados. correlações foram achadas diferenças significativas entre o desempenho de ambos grupos das crianças avaliadas, e uma correlaçÃo significativa entre o ICV e subdomínio de compreensÃo de leitura da ENI. ConclusÃo. Estes resultados estÃo acordes com a hipótese de que o desenvolvimento da linguagem oral é um dos principais facilitadores a aquisiçÃo da aprendizagem da leitura, posto que na medida em que a criança compreende a semântica oral estará melhor preparada para dotar de sentido o que lê.

Immunocompetent truncated E2 glycoprotein of bovine viral diarrhea virus (BVDV) expressed in Nicotiana tabacum plants: a candidate antigen for new generation of veterinary vaccines.

The bovine viral diarrhea virus (BVDV) is the etiological agent responsible for a wide spectrum of clinical diseases in cattle. The glycoprotein E2 is the major envelope protein of this virus and the strongest inductor of the immune response. There are several available commercial vaccines against bovine viral diarrhea (BVD), which show irregular performances. Here, we report the use of tobacco plants as an alternative productive platform for the expression of the truncated version of E2 glycoprotein (tE2) from the BVDV. The tE2 sequence, lacking the transmembrane domain, was cloned into the pK7WG2 Agrobacterium binary vector. The construct also carried the 2S2 Arabidopsis thaliana signal for directing the protein into the plant secretory pathway, the Kozak sequence, an hexa-histidine tag to facilitate protein purification and the KDEL endoplasmic reticulum retention signal. The resulting plasmid (pK-2S2-tE2-His-KDEL) was introduced into Agrobacterium tumefaciens strain EHA101 by electroporation. The transformed A. tumefaciens was then used to express tE2 in leaves of Nicotiana tabacum plants. Western blot and ELISA using specific monoclonal antibodies confirmed the presence of the recombinant tE2 protein in plant extracts. An estimated amount of 20 µg of tE2 per gram of fresh leaves was regularly obtained with this plant system. Injection of guinea pigs with plant extracts containing 20 µg of rtE2 induced the production of BVDV specific antibodies at equal or higher levels than those induced by whole virus vaccines. This is the first report of the production of an immunocompetent tE2 in N. tabacum plants, having the advantage to be free of any eventual animal contaminant.

Expression of the antibody 14D9 in Nicotiana tabacum hairy roots

Nicotiana tabacum hairy roots that express the antibody 14D9 were established. The 14D9 antibody yield obtained after 20 days of culture was 5.95 μg 14D9ml-1. The addition of the reticulum endoplasmic retention sequence KDEL demonstrated a positive effect over the intracellular 14D9 amounts with a yield increase up to 20.82 µg ml-1. DMSO increased the antibody amount in the biomass from 20.00 to 64.03 µg ml-1 while PVP (at 1.5 gl-1) and gelatine (at 5.0 gl-1) increased total 14D9 amounts in the culture medium to 25 µg and 14 µg respectively.