Systematic study of the genetic response of a variable virus to the introduction of deleterious mutations in a functional capsid region.

We have targeted the intersubunit interfaces in the capsid of foot-and-mouth disease virus to investigate the genetic response of a variable virus when individual deleterious mutations are systematically introduced along a functionally defined region of its genome. We had previously found that the individual truncation (by mutation to alanine) of 28 of the 42 amino acid side chains per protomer involved in interactions between capsid pentameric subunits severely impaired infectivity. We have now used viral RNAs individually containing each of those 28 deleterious mutations (or a few others) to carry out a total of 96 transfections of susceptible cells, generally followed by passage(s) of the viral progeny in cell culture. The results revealed a very high frequency of fixation in the capsid of second-site, stereochemically diverse substitutions that compensated for the detrimental effect of primary substitutions at many different positions. Most second-site substitutions occurred at or near the capsid interpentamer interfaces and involved residues that are spatially very close to the originally substituted residue. However, others occurred far from the primary substitution, and even from the interpentamer interfaces. Remarkably, most second-site substitutions involved only a few capsid residues, which acted as "second-site hot spots." Substitutions at these hot spots compensated for the deleterious effects of many different replacements at diverse positions. The remarkable capacity of the virus to respond to the introduction of deleterious mutations in the capsid with the frequent fixation of diverse second-site mutations, and the existence of second-site hot spots, may have important implications for virus evolution.

Engineering viable foot-and-mouth disease viruses with increased thermostability as a step in the development of improved vaccines.

We have rationally engineered foot-and-mouth disease virus to increase its stability against thermal dissociation into subunits without disrupting the many biological functions needed for its infectivity. Amino acid side chains located near the capsid intersubunit interfaces and either predicted or found to be dispensable for infectivity were replaced by others that could establish new disulfide bonds or electrostatic interactions between subunits. Two engineered viruses were normally infectious, genetically stable, and antigenically indistinguishable from the natural virus but showed substantially increased stability against irreversible dissociation. Electrostatic interactions mediated this stabilizing effect. For foot-and-mouth disease virus and other viruses, some evidence had suggested that an increase in virion stability could be linked to an impairment of infectivity. The results of the present study show, in fact, that virion thermostability against dissociation into subunits may not be selectively constrained by functional requirements for infectivity. The thermostable viruses obtained, and others similarly engineered, could be used for the production, using current procedures, of foot-and-mouth disease vaccines that are less dependent on a faultless cold chain. In addition, introduction of those stabilizing mutations in empty (nucleic acid-free) capsids could facilitate the production of infection-risk-free vaccines against the disease, one of the economically most important animal diseases worldwide.

Thermostable variants are not generally represented in foot-and-mouth disease virus quasispecies.

A severe limitation to fully realize the dramatic potential for adaptation of RNA virus quasispecies may occur if mutations in vast regions of the sequence space of virus genomes lead to significant decreases in biological fitness. In this study the detection and selection by heat of thermostable variants from different foot-and-mouth disease virus (FMDV) populations were attempted, in order to explore whether FMDV may generally accept a substantial increase in thermostability without compromising its infectivity. The results obtained with both uncloned and cloned populations of different serotypes, recovered from cytolytic or persistent infections and subjected to either very few passages or extensive passaging in cells, indicate that the presence of thermostable virus variants, even in small proportions, is not a general feature of FMDV quasispecies. This suggests that no substantial increase in the thermostability of FMDV may readily occur without a negative effect on viral function.

Determinación de emisión de dióxido de carbono (Co2) y el impacto económico y medioambiental en la construcción de viviendas de interés social de El Salvador: aplicación en sector construcción / Determination of carbon dioxide (Co2) emission and the economic and environmental impact in the construction of low-income housing in El Salvador: application in the construction sector

El objetivo de este proyecto fue crear un informe técnico del valor económico y el impacto ambiental del dióxido de carbono (CO2), que genera la construcción de las viviendas de interés social en El Salvador. Se procedió a reconocer terminología técnica sobre los diferentes factores que intervienen en el análisis de la relación de costos económicos y medioambientales. Luego se identificaron los modelos de cuantificación de Emisiones de CO2 para construcciones. Finalmente se creó una herramienta para el levantamiento de información de tipo cualitativa y cuantitativa de viviendas y para la obtención de datos técnicos. De acuerdo con lo investigado, se encontró que los materiales de construcción con mayores emisiones de CO2 por kilogramo son el acero y el hormigón armado utilizado en las partes estructurales, así como el vidrio y losas para las envolventes, seguidos de la madera y el cemento, siendo el menor el hormigón premezclado, ladrillos rojos y arena.

The objective of this project was to create a technical report of the economic value and the environmental impact of carbon dioxide (CO2), generated by the construction of social housing in El Salvador. We proceeded to recognize technical terminology on the different factors involved in the analysis of the relationship between economic and environmental costs. Then the quantification models of CO2 Emissions for constructions were identified. Finally, a tool was created to collect qualitative and quantitative information on housing and to obtain technical data. According to what was investigated, it was found that the construction materials with the highest CO2 emissions per kilogram are steel and reinforced concrete used in the structural parts, as well as glass and slabs for the envelopes, followed by wood and cement, the least being pre-mixed concrete, red bricks and sand.

Diseño innovador para remodelación arquitectónica y estudio de eficiencia energética en áreas de rehabilitación integral de la Fundación Teleón Pro Rehabilitación: en asocio con FUNTER / Innovative design for remodeling architectural and efficiency study energy in rehabilitation areas of Teletón Foundation Pro Rehabilitation: in association with FUNTER

Se desarrolló una propuesta de intervención arquitectónica y constructiva de iluminación natural, ventilación y confort térmico para el área de rehabilitación del edificio de la Fundación Teletón Pro Rehabilitación FUNTER. Se diseñó una propuesta de interiorismo para uso eficiente de espacios, mobiliario, iluminación artificial, texturas y colores apropiados para el desarrollo de terapias. Se realizó un estudio de eficiencia energética para proponer medidas de ahorro y uso eficiente de la energía en el edificio. Se diseñó una nueva distribución de espacios en la zona de terapia y se aplicaron criterios de bioclimatismo pasivo, como la ventilación cruzada, ventilación cenital y la apertura de áreas de ventilación basándose en el análisis de vientos del sitio para mejorar la sensación térmica interna. La implementación de un jardín sensorial ayudará a los pacientes de terapia de marcha a interactuar en un área más cercana a las superficies reales del ambiente.

An architectural and constructive intervention proposal for natural lighting, ventilation and thermal comfort was developed for the rehabilitation area of ​​the Teletón Pro Rehabilitation Foundation FUNTER building. An interior design proposal was designed for the efficient use of spaces, furniture, artificial lighting, textures and appropriate colors for the development of therapies. An energy efficiency study was carried out to propose measures for saving and efficient use of energy in the building. A new distribution of spaces was designed in the therapy area and passive bioclimatic criteria were applied, such as cross ventilation, overhead ventilation and the opening of update areas in the site wind analysis to improve the internal thermal sensation. Implementing a sensory garden will help gait therapy patients interact in an area closer to the actual surfaces of the environment.

Vitrificación de embriones y transferencia en ciclo posterior como estrategia segura y eficiente en pacientes altas respondedoras / No disponible

OBJETIVO: El objetivo de este estudio retrospectivo fue comparar el impacto de dos estrategias de maduración ovocitaria en un grupo de pacientes alta respondedoras con riesgo de hiperestimulación ovárica. MATERIAL Y MÉTODOS: La pauta de estimulación empleada, basada en el empleo concomitante de la hormona folículo estimulante (FSHr) (75 IU) y la gonadotropina menopaúsica humana (hMG-HP) (75 IU), consistió en un protocolo corto con antagonistas de la hormona liberadora de gonadotropinas (GnRH) en combinación con dos pautas de maduración ovocitaria: administración de un bolo de agonista de la GnRH (aGnRH) (n=37) seguida de la criopreservación de embriones y transferencia embrionaria en ciclo posterior o de gonadotropina coriónica humana (hCG) (n=49) y transferencia embrionaria en fresco. RESULTADOS: Los resultados obtenidos fueron comparables para la tasa de fecundación siendo significativamente superior en el grupo donde se administró el aGnRH como inductor de la ovulación (739/888 [83,2%] vs. 441/618 [71,3%], p < 0,05). No se obtuvieron diferencias significativas para las tasas de implantación (35/100 [35%] vs. 41/127 [32,3%]) o embarazo clínico (30/36 [83,3%] vs. 32/41 [78%]). CONCLUSIÓN: La estrategia conseguida al combinar la administración de un bolo de aGnRH con la vitrificación programada de embriones ofrece una alternativa segura y eficiente para evitar la hiperestimulación ovárica sin restar posibilidades de embarazo. La inducción de la maduración ovocitaria con agonistas de la GnRH en pacientes con alta respuesta consigue la obtención de un mayor número de embriones para realizar una transferencia embrionaria sin comprometer el estado clínico de la paciente ni los resultados del ciclo

OBJECTIVE: The aim of this retrospective study was to compare the impact of two strategies for oocyte maturation in a group of high responders at risk of ovarian hyperstimulation. MATERIALS AND METHODS: The stimulation based on a concomitant administration of recombinant follicle stimulating hormone (rFSH) (75 UI) and human menopausal gonadotrophin (hMG-HP) (75 UI), was arranged as a gonadotrophin releasing hormone (GnRH) antagonist short protocol combined with two different approaches of final oocyte maturation: GnRH agonist triggering (aGnRH) and embryo transfer in a later non stimulated cycle (n=37) or human chorionic gonadotropin (hCG) followed by fresh embryo transfer (n=49). RESULTS: The fertilization rate resulted significantly higher when aGnRH was used as ovulation inducer (739/888 [83.2%] vs. 441/618 [71.3%], p <0.05). No differences between the subgroups were detected in terms of cumulative implantation (35/100 [35%] vs. 41/127 [32.3%]) and clinical pregnancy rates (30/36 [83.3%] vs. 32/41 [78%]). CONCLUSION: The strategy achieved by combining the administration of a single bolus of aGnRH with elective vitrification of all embryos offers a safe and efficient alternative to avoid the risk of ovarian hyperstimulation without decreasing the chances of pregnancy. The induction of oocyte maturation with GnRH agonists in high responders provides a high number of embryos to be transferred without compromising the clinical condition of the patient and reproductive outcome