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Human Embryonic Kidney cells (HEK293) are a popular host for recombinant protein expression and production in the biotechnological industry. This has driven within both, the scientific and the engineering communities, the search for strategies to increase their protein productivity. The present work is inserted into this search exploring the impact of adding sodium acetate (NaAc) into a batch culture of HEK293 cells. We monitored, as a function of time, the cell density, many external metabolites, and the supernatant concentration of the heterologous extra-cellular domain ECD-Her1 protein, a protein used to produce a candidate prostate cancer vaccine. We observed that by adding different concentrations of NaAc (0, 4, 6 and 8 mM), the production of ECD-Her1 protein increases consistently with increasing concentration, whereas the carrying capacity of the medium decreases. To understand these results we exploited a combination of experimental and computational techniques. Metabolic Flux Analysis (MFA) was used to infer intracellular metabolic fluxes from the concentration of external metabolites. Moreover, we measured independently the extracellular acidification rate and oxygen consumption rate of the cells. Both approaches support the idea that the addition of NaAc to the culture has a significant impact on the metabolism of the HEK293 cells and that, if properly tuned, enhances the productivity of the heterologous ECD-Her1 protein.

Desde los inicios de la ocupación española en el Perú en el siglo XVI y tras el proceso de transculturización, una gran variedad de muros, arcos y bóvedas se construyeron con tamaños similares a los ejecutados en la península Ibérica. Al inicio, estas obras fueron hechas con ladrillo, piedra y adobe; sin embargo, con los frecuentes sismos y terremotos que hicieron colapsar gran parte de estas edificaciones, se evidenció la falta de previsión constructiva de los alarifes españoles. Por tanto, se tuvo que recurrir a la quincha, material hecho de entramados de madera rolliza, caña, y cubierta de barro que fuera utilizado en la construcción de viviendas y centros administrativos prehispánicos. La presente investigación muestra los resultados de un trabajo de campo, descriptivo, de revisión bibliográfica completa que tuvo como objetivo demostrar el origen y evolución de las bóvedas de quincha en el Perú; la capacidad sismorresistente del material, así como la eficacia estructural que tuvo en la construcción de las bóvedas y cúpulas de las principales iglesias de Trujillo. Los resultados permiten una mayor comprensión de la arquitectura virreinal y evidencian su utilidad ancestral ante el contexto moderno, así como su estética cautivadora que permite la identificación del trujillano con su patrimonio cultural.

La urgente necesidad de desarrollar y producir vacunas seguras y efectivas para garantizar la reducción de la propagación del coronavirus de tipo 2 causante del síndrome respiratorio agudo severo, hizo que el Centro de Inmunología Molecular y el Instituto Finlay de Vacunas, desarrollaran dos vacunas y un candidato vacunal contra la COVID-19, que tienen como componente la molécula del dominio de unión al receptor (aa 319-541) del virus. Para establecer el proceso productivo, se realizaron experimentos en los posibles pasos del proceso de purificación de la molécula del dominio de unión al receptor (aa 319-541), con vistas a su posterior transferencia tecnológica a escala industrial. Dicha molécula está fusionada con una etiqueta de hexahistidina en su extremo C-terminal y presenta nueve residuos de cisteína en su secuencia que forman cuatro enlaces disulfuros intramoleculares, quedando una cisteína libre que permite obtener dos moléculas: dimérica y monomérica, antígenos que forman parte de las vacunas SOBERANA®02 y SOBERANA®Plus y el candidato vacunal SOBERANA 01. Se determinaron las mejores condiciones de adsorción de las matrices cromatográficas de afinidad por quelatos metálicos, intercambio catiónico y exclusión molecular. Se evaluó el desempeño del proceso a escala piloto y se caracterizó la molécula de acuerdo a sus propiedades físico-químicas y biológicas. Los resultados obtenidos mostraron un 60,02 ± 5,15% de recuperación total de la proteína de interés, con más del 98% de pureza en ambas moléculas, una eficiente remoción de contaminantes y una antigenicidad mayor del 90% referido al monómero control del dominio de unión al receptor con 99% de pureza, lo que demuestra que el proceso establecido es eficiente en la obtención de un producto con la calidad requerida.

The genus Swietenia includes two mahogany species, Swietenia macrophylla (King.), commonly known as Honduran mahogany, and Swietenia mahagoni (L.) Jacq, commonly known as Cuban mahogany. There are reports of morphologically intermediate forms between Cuban and Honduran mahogany that have been localized in some Caribbean islands. The main objective of this research is to distinguish morphologically intermediate forms between the parental species S. mahagoni and S. macrophylla based on leaf morphological characters. Phenotypic data from a total of 357 mahogany trees were collected in the province of Sancti Spiritus, Cuba. Rachis length, petiole length, leaflet length, leaflet width, number of leaflets and number of nerves were evaluated. Morphological descriptor analysis revealed significant differences in the morphometric variables of the evaluated leaves, with rachis length, number of leaflets, leaflet length, petiole length, leaflet width, and number of nerves being significantly higher in S. macrophylla than in S. mahagoni. ANOVA reflected the variance between all leaf morphological parameters evaluated between the two pure species with statistically significant difference. Using the function obtained in the first DC of the two species, the individuals in the mixed stands were classified into 71 morphologically intermediate forms, 45 S. macrophylla and 64 S. mahagoni. In the second DC with the two species and the morphologically intermediate forms, Wilks’ partial lambda indicates that the variable "leaflet length" with 0.67 contributed most to the overall discrimination.

Habitual consumption of highly palatable foods when not in metabolic need (HPF eating) is linked to obesity. High HPF consumption is also linked to mental health disorder (MHD) symptoms. Mindfulness-based interventions are popular treatments for obesity and MHDs, but little is known about the relationship between trait mindfulness and motive-based HPF eating. Therefore, a total of 927 young adults completed a survey that included the Palatable Eating Motives Scale-7 (which identifies Coping-, Reward enhancement-, Social-, and Conformity-eating), the Mindful Attention Awareness Scale, the Perceived Stress Scale, and demographic and body mass index (BMI) questions. An MHD questionnaire allowed a comparison of HPF eating between participants with and without various MHDs. Regressions revealed that Coping-eating was independently associated with lower mindfulness and also greater perceived stress, higher BMI, and female sex. Of these variables, only lower mindfulness was independently associated with Reward-, Social-, and Conformity-eating. Coping- and Reward-eating were more frequent in participants with versus without an anxiety disorder, depression, ADD/ADHD, and PTSD. Coping-eating was also more frequent in participants with body dysmorphic disorder. These findings warrant investigations in participants with clinically validated diagnoses for DSM-specific MHDs. Results from such investigations and the uncovered nature of associations between motive-specific HPF eating and trait mindfulness could provide novel targets to improve mindfulness-based interventions for obesity and MHDs.

Abstract The COVID-19 pandemic has caused a large number of diseases worldwide. There are few vaccines to constrain this disease and the value of them is high. In this sense, the antigens of the vaccine platform Soberana, the receptor binding domain from SARS-CoV-2 Spike protein, both the monomeric (mRBD) and dimeric (dRBD) forms, have been developed. This study encompassed several analyses by different techniques like circular dichroism (CD), fluorescence spectroscopy (FS) and Gel Filtration- High Performance Liquid ChLC of mRBD and dRBD. Monomer and dimer exhibited similar far-UV CD spectral characteristics with 54% of β-sheet content. Similar conformational features according to near-UV CD and FS studies were observed in both RBD. Stress stability studies by far-UV CD, FS, biological activity and GF-HPLC at 37 °C showed that mRBD is very stable. On the other hand, dRBD fluorescent emission showed a shift towards higher wavelengths as the incubation time increases, suggesting exposition of tryptophan residues, unlike what happens with mRBD. Biological activity outcome confirms these results. GF-HPLC profiles showed that in mRBD, the product of molecular stress are dimers and does not increase over time. However, dRBD showed dimer fragmentation as the main degradation species. This study reveals the usefulness of CD techniques for the analysis of degradation of RBD molecules as well as showed the difference in stability of both RBD molecules. Besides, our work provides useful insights into the production of a key protein used in diagnosis and therapeutics to fight COVID-19 pandemia.

We have developed a single process for producing two key COVID-19 vaccine antigens: SARS-CoV-2 receptor binding domain (RBD) monomer and dimer. These antigens are featured in various COVID-19 vaccine formats, including SOBERANA 01 and the licensed SOBERANA 02, and SOBERANA Plus. Our approach involves expressing RBD (319-541)-His6 in Chinese hamster ovary (CHO)-K1 cells, generating and characterizing oligoclones, and selecting the best RBD-producing clones. Critical parameters such as copper supplementation in the culture medium and cell viability influenced the yield of RBD dimer. The purification of RBD involved standard immobilized metal ion affinity chromatography (IMAC), ion exchange chromatography, and size exclusion chromatography. Our findings suggest that copper can improve IMAC performance. Efficient RBD production was achieved using small-scale bioreactor cell culture (2 L). The two RBD forms - monomeric and dimeric RBD - were also produced on a large scale (500 L). This study represents the first large-scale application of perfusion culture for the production of RBD antigens. We conducted a thorough analysis of the purified RBD antigens, which encompassed primary structure, protein integrity, N-glycosylation, size, purity, secondary and tertiary structures, isoform composition, hydrophobicity, and long-term stability. Additionally, we investigated RBD-ACE2 interactions, in vitro ACE2 recognition of RBD, and the immunogenicity of RBD antigens in mice. We have determined that both the monomeric and dimeric RBD antigens possess the necessary quality attributes for vaccine production. By enabling the customizable production of both RBD forms, this unified manufacturing process provides the required flexibility to adapt rapidly to the ever-changing demands of emerging SARS-CoV-2 variants and different COVID-19 vaccine platforms.

The data for provide evidences of the multi steady state of the human cell line HEK 293 was obtained from 2 L bioreactor continuous culture. A HEK 293 cell line transfected to produce soluble HER1 receptor was used. The bioreactor was operated at three different dilution rates in sequential manner. Daily samples of culture broth were collected, a total of 85 samples were processed. Viable cell concentration and culture viability was addressing by trypan blue exclusion method using a hemocytometer. Heterologous HER1 supernatant concentration was quantified by a specific ELISA and the metabolites by mass spectrometry coupled to a liquid chromatography.The primary data were collected in excel files, where it was calculated the kinetic and other variables by using mass balance and mathematical principles. It was compared the steady states behavior each other's to find out the existence of steady states’ multiplicity, taking into account the stationary phase with respect to the cell density (which means its coefficient of variation is less than 20 %).From the metabolic measurements by using Liquid Chromatography coupled to mass spectrometry (LC-MS), it was also built the data matrix with the specific rates of the 76 metabolites obtained. The data were processed and analyzed, using multivariate data asssnalysis (MVDA) to reduce the complexity and to find the main patterns present in the data.We describe also the full data of the metabolites not only for steady states but also in the time evolution, which could help others in terms of modeling and deep understanding of HEK293 metabolism, especially under different culture conditions.

Use of irrigation in forage production systems based on perennial C4 grasses has been limited because of inconsistent responses of biomass production to water. The effects of three different levels of irrigation on growth of Paspalum notatum (Bahia grass) and P. dilatatum (dallis grass) over two consecutive growing seasons were studied. The phenological state of the plants was inferred by the proportion of lamina and non-lamina components. While irrigation failed to increase annual dry matter yield of forage, phenological composition of forage was affected by irrigation with both species showing changes in distribution of dry matter accumulation among different above-ground plant parts. Future research on these species should study the effects of moisture deficit on phenology to better understand the effects of irrigation.

Introduction Acute radiation syndrome (ARS) is a high-risk, low-frequency diagnosis that can be fatal and is difficult to diagnose without an obvious history of ionizing radiation exposure. Methods Twenty-two emergency medicine residents and one pharmacy resident participated in an hour-long simulation session. To accommodate all learners, the simulation was conducted eight times over a block of scheduled time (two to four learners/session). Sessions included a prebriefing, pre/post questionnaires, the ARS case, and a debriefing. Learners evaluated and managed a 47-year-old male (manikin) with the hematopoietic and cutaneous subsyndromes of ARS who presented with hand pain/erythema/edema and underlying signs of infection 2 weeks after an unrecognized radiation exposure. Learners had to perform a history and physical, recognize/manage abnormal vitals, order/interpret labs, consult appropriate disciplines, and initiate supportive care. Results There was a mean reported increase in ability to recognize signs and symptoms of ARS (p < .001) and appropriately manage a patient with this condition (p = .03) even after controlling for baseline confidence in ability to make and manage uncommon diagnoses, respectively. Learners rated this simulation as a valuable learning experience, effective in teaching them how to diagnose and treat ARS, and one they would recommend to other health care professionals. Discussion This simulation aimed to teach the diagnosis and initial management of the hematopoietic and cutaneous subsyndromes of ARS. It should be used to increase awareness of the potential for ionizing radiation exposure under less obvious conditions and raise the index of suspicion for ARS in the undifferentiated patient.

Durante su vida universitaria, el estudiante de arquitectura realiza diversos trabajos manuales que son imprescindibles en su formación académica, como la elaboración de maquetas, planos e informes que, en su mayoría, son posteriormente desechados sin considerar su valorización. La presente investigación tuvo como objetivo el análisis de la gestión de los residuos sólidos generados en el Programa de estudio de Arquitectura de la Facultad de Arquitectura, Urbanismo y Artes (FAUA) de la Universidad Privada Antenor Orrego (UPAO), considerando los principios de las 5R y la cultura de reciclaje que los alumnos poseen. Para ello, primero se caracterizaron los residuos producidos según su tiempo de vida y toxicidad. Luego se aplicaron 218 encuestas a alumnos escogidos bajo muestreo probabilístico aleatorio simple con el fin de conocer sus hábitos ecológicos y su predisposición para el reciclaje, logrando una estimación de la cantidad de materiales utilizados y desechados durante el ciclo académico; asimismo, se realizó una entrevista a la directiva para tener conocimiento de la gestión actual. Finalmente, se propuso un plan integral de residuos para la Facultad, que incluyó la implementación de un área de reciclaje dentro del pabellón, el cual permitirá organizar adecuadamente los residuos sólidos promoviendo una cultura ambiental en el estudiante, plana docente y administrativa; todo con el fin de llevar a la práctica el respeto por el medio ambiente, en pro de una mejora en la utilización de recursos y valorización de residuos, contribuyendo así a la huella ecológica.

El presente estudio tiene por objetivo determinar la necesidad de incorporar una perspectiva de género en la política de perfeccionamiento doctoral de la Facultad de Ciencias Jurídicas de la Universidad de Antofagasta. Para tal efecto, se estudió la situación de las mujeres en la academia a nivel general, pudiendo evidenciar que existe una brecha de género en las universidades a nivel mundial. Bajo dicho contexto, se delimitó el objetivo de esta investigación en el proceso de selección del profesorado de la Facultad de Ciencias Jurídicas para proseguir programas de doctorado. Se constató que la Universidad de Antofagasta desde sus primeros planes de desarrollo declaró como una de sus fortalezas contar con académicos/as doctores/as, fijando estrategias, objetivos e indicadores para incrementarlos, y reguló, a contar del 2002, el acceso a los estudios de postgrado, fijando el proceso y criterios de selección de los académico/as. Dado ello, se revisó la implementación de la política de perfeccionamiento doctoral en el Departamento de Derecho y en la Facultad de Ciencias Jurídicas, constatándose que no existe un enfoque de género en la selección de los académicos/as que han proseguido estudios doctorales, al ser sólo varones los beneficiados, por lo que se propone como mecanismo para disminuir dicha brecha la incorporación de la perspectiva de género en la política de perfeccionamiento de dichas unidades académicas.

El diseño e implementación de sistemas informáticos para las organizaciones deben basarse en las políticas y reglas del negocio. Hoy día se considera ventajoso aplicar el enfoque de reglas de negocio en el desarrollo de sistemas de información, y de esta manera garantizar la inserción y modificación de las reglas de manera automática. Una de las formas para implementar un sistema de información usando reglas de negocio, es a través del tipo de herramienta independiente de los gestores de datos, pero que genera recursos de bases de datos. Ante la necesidad de desarrollar un sistema de información transaccional para la gestión de menús de comedores en la UCLV, se hizo necesaria una aplicación cuya interfaz de entrada realice las operaciones de inserción y actualización en una base de datos. La existencia de un conjunto de reglas de negocio que deben ser chequeadas al realizar estas operaciones, hace posible la utilización de la herramienta LPT SQL con vistas a disminuir el esfuerzo de programación al insertar el código, que garantiza el cumplimiento de las reglas, y su posible modificación independientemente de la interfaz de usuario.

Many of the studies in Campos grasslands focus on management aspects such as the control of herbage allowance, and application of nutrients and/or overseeding with legumes. However, there is little literature on how the Campos grassland resource is utilised, especially regarding the grazing pattern and the relationship between pasture quantity and quality on daily grazing activities. The study of the ingestive behaviour in species-rich and heterogeneous native grasslands during daylight hours, and understanding how animals prioritise quality or quantity of intake in relation to pasture attributes, are important to comprehend the ingestive-digestive processes modulating the energy intake of animals and to achieve a better grazing management. Therefore, the objective was to describe and quantify the daily grazing behaviour of growing cattle grazing native pasture with different structures as a result of different management practices, and study the relationship of pasture attributes and intake through multivariate analysis. The study was carried out at the Faculty of Agronomy, Paysandú, Uruguay. Treatments were native grassland, overseeding with Trifolium pratense and Lotus tenuis + phosphorus, and native pasture + nitrogen-phosphorus. Grazing activities were discriminated into grazing, searching (defined when animals take 1–2 bites in one feeding station and then change to another feeding station and so on), ruminating and idling. The probability of time allocated to each activity was continuously measured during daylight hours (0700–1930) and was related to pasture structure and forage quality using regression tree models, while the bite rate was determined every 2 h. The diurnal pattern of growing cattle showed grazing and searching sessions, followed by ruminating and idling sessions. The length of sessions (as the probability of time allocated to each activity) varied throughout the day. The grazing probability was greater during afternoon than morning and midday (0.74 vs 0.45 vs 0.46, respectively), and it was associated with higher bite rate (34.2 bites/min). Regression tree models showed different grazing, searching and ruminating strategies according to pasture attributes. During the morning, animals modified grazing, searching, ruminating and idling strategies according to bite rate, crude protein in diet and herbage allowance. At midday, they only adjusted ruminating and idling, while during afternoon sessions, grazing activities were modified by pasture quantity attributes such as herbage mass and herbage allowance. By controlling the herbage allowance, herbage mass and pasture height, animals prioritise quality in the morning and quantity in the afternoon, integrating and modifying the grazing-searching and ruminating-idling pattern.