Rational design for the recombinant expression of the Receptor Binding Domain of SARS-CoV-2 Spike Glycoprotein

Background: COVID-19 represents a significant threat to global human health. SARS-CoV-2, the etiologic viral agent, needs to be under-covered at the structural biology level to facilitate the rational design of diagnostic tests and vaccine candidates. SARS-CoV-2 Receptor Binding Domain of Spike protein (RBD-S) acts as the key to open the gate, to enter the cells during infection. Thus, it is a stronger candidate for designing effective antigens for vaccines and diagnostics. Here, we relied on the viral DNA codifying to RBD-S to use synthetic biology for optimizing the recombinant expression of this (rRBD-S) as a proof of concept of rational designs of bioprocess for vaccine candidates and immunogens to improved rapid diagnostic tests. Methods: rRBD-S coding sequences inspired on RBD-S ectodomain from SARS-CoV-2 were designed, codon-optimized, tagged, synthesized, cloned in an expression vector (pD444-MR), and transformed into C41(DE3)pLysS E. coli strain. Expression of recombinant RBD-S was resulting in a protein purified using Ni-IMAC (Nickel Immobilized metal affinity chromatography). Results: rRBD-S produced result in a ~30KDa protein with yields of 4.618 gr L-1. Protein was recovered from the bacterial soluble fraction without refolding process. Conclusions: rRBD-S is an important tool for immunity diagnostics as Lateral-Flow-Devices, structural biology studies, and even as vaccine candidate for combating SARS-CoV-2. Notably considering the advantages of rational subunit vaccines for immune response against other vaccines technologies whose effectiveness in the long-term process has not been demonstrated yet. Acknowledgements: We thank HIDALGO´s Council of Science, Technology and Innovation (CITNOVA) for the GRANT 20201122 to LMRM

Comparison of Mechanical and Structural Properties of Nickel-titanium Alloy with Titanium-molybdenum Alloy and Titanium-niobium Alloy as Potential Metals for Endodontic Files

Introduction: The objective of this study was to compare the mechanical and structural properties of the nickel-titanium (Ni-Ti) alloy already used in endodontics with titanium-molybdenum (Ti-Mo) and titanium-niobium (Ti-Nb) alloys to determine if these can be suggested in the manufacture of endodontic files. Methods and Materials: Orthodontic wires made of the different alloys were used. The previously mentioned alloys were characterized by energy-dispersive X-ray spectroscopy (EDX), X-ray diffraction (XRD) and torsion tests. Cyclic fatigue tests were performed on a simulated canal with a curvature of 86° to 375 rpm. The fractured surfaces of the wires were observed by means of scanning electron microscopy (SEM). A Kruskal-Wallis test and U Mann Whitney test were used to determine significant differences in cyclic fatigue between groups. Results: In the mechanical tests, similar values of torsion were found for the three alloys. In XRD, the Ti-Nb showed less structural changes. In the cyclic fatigue test, Ti-Nb was found to be significantly more resistant with respect to Ni-Ti and Ti-Mo. Conclusion: Based on our in vitro study, Ti-Nb is suggested as a possible alloy for the manufacture of rotary files due to its impressive properties

P3-062: Wood-smoke exposure as a survival predictor in non-small cell lung cancer with response to erlotinib: an open label phase II study

This paper explores the opportunity for New Zealand to establish and sustain an internationally competitive sheep dairy industry. As part of this it evaluates the role of responsible innovation (RI) within the New Zealand sheep dairy (NZSD) industry and whether this can assist in achieving a sustainable competitive advantage. In the context of agrifood supply chains RI has received little attention despite the fact that these industries have significant environmental, ethical and social impacts. The research also addresses the lack of evidence as how to put RI into practice and the claim that the practical applicability of RI is not possible. The French sheep dairy industry was used as a comparative case study for the New Zealand industry. Information was gathered through a literature search, the comparative case study and interviews with New Zealand and French industry experts. Comparisons were made between the strategic capabilities and structural forces of the sheep dairy industries in both France and New Zealand. The study found that for the NZSD industry to achieve a competitive advantage it would need to pursue a differentiation strategy that focused on customer responsiveness, innovation, sustainability and quality. Furthermore, the study identified that RI had the potential to assist the NZSD industry by providing distinctive competencies to develop a competitive advantage. This is because there were existing resources and capabilities that provided a platform for differentiation. There were also strategic and economic drivers in the NZSD industry that encouraged RI as a competitive strategy. This indicated that for RI to occur there needed to be economic incentives that encouraged companies to pursue this strategy

Cell-Mediated Immunity Generated in Response to a Purified Inactivated Vaccine for Dengue Virus Type 1

Dengue is the most prevalent arboviral disease afflicting humans, and a vaccine appears to be the most rational means of control. Dengue vaccine development is in a critical phase, with the first vaccine licensed in some countries where dengue is endemic but demonstrating insufficient efficacy in immunologically naive populations. Since virus-neutralizing antibodies do not invariably correlate with vaccine efficacy, other markers that may predict protection, including cell-mediated immunity, are urgently needed. Previously, the Walter Reed Army Institute of Research developed a monovalent purified inactivated virus (PIV) vaccine candidate against dengue virus serotype 1 (DENV-1) adjuvanted with alum. The PIV vaccine was safe and immunogenic in a phase I dose escalation trial in healthy, flavivirus-naive adults in the United States. From that trial, peripheral blood mononuclear cells obtained at various time points pre- and postvaccination were used to measure DENV-1-specific T cell responses. After vaccination, a predominant CD4+ T cell-mediated response to peptide pools covering the DENV-1 structural proteins was observed. Over half (13/20) of the subjects produced interleukin-2 (IL-2) in response to DENV peptides, and the majority (17/20) demonstrated peptide-specific CD4+ T cell proliferation. In addition, analysis of postvaccination cell culture supernatants demonstrated an increased rate of production of cytokines, including gamma interferon (IFN-γ), IL-5, and granulocyte-macrophage colony-stimulating factor (GM-CSF). Overall, the vaccine was found to have elicited DENV-specific CD4+ T cell responses as measured by enzyme-linked immunosorbent spot (ELISpot), intracellular cytokine staining (ICS), lymphocyte proliferation, and cytokine production assays. Thus, together with antibody readouts, the use of a multifaceted measurement of cell-mediated immune responses after vaccination is a useful strategy for more comprehensively characterizing immunity generated by dengue vaccines

The effect of orthodontic forces on calcitonin gene-related peptide (CGRP) expression in the human periodontal ligament and its relationship with the human dental pulp

The purpose of this study was to quantify the effect of moderate and severe orthodontic forces on Calcitonin gene-related peptide (CGRP) expression in the healthy human periodontal ligament (PDL) and its possible relationship with the human dental pulp

Genomic diversity of human papillomavirus-16, 18, 31, and 35 isolates in a Mexican population and relationship to European, African, and Native American variants

AbstractCervical cancer, mainly caused by infection with human papillomaviruses (HPVs), is a major public health problem in Mexico. During a study of the prevalence of HPV types in northeastern Mexico, we identified, as expected from worldwide comparisons, HPV-16, 18, 31, and 35 as highly prevalent. It is well known that the genomes of HPV types differ geographically because of evolution linked to ethnic groups separated in prehistoric times. As HPV intra-type variation results in pathogenic differences, we analyzed genomic sequences of Mexican variants of these four HPV types. Among 112 HPV-16 samples, 14 contained European and 98 American Indian (AA) variants. This ratio is unexpected as people of European ethnicity predominate in this part of Mexico. Among 15 HPV-18 samples, 13 contained European and 2 African variants, the latter possibly due to migration of Africans to the Caribbean coast of Mexico. We constructed phylogenetic trees of HPV-31 and 35 variants, which have never been studied. Forty-six HPV-31 isolates from Mexico, Europe, Africa, and the United States (US) contained a total of 35 nucleotide exchanges in a 428-bp segment, with maximal distances between any two variants of 16 bp (3.7%), similar to those between HPV-16 variants. The HPV-31 variants formed two branches, one apparently the European, the other one an African branch. The European branch contained 13 of 29 Mexican isolates, the African branch 16 Mexican isolates. These may represent the HPV-31 variants of American Indians, as a 55% prevalence of African variants in Mexico seems incomprehensible. Twenty-seven HPV-35 samples from Mexico, Europe, Africa, and the US contained 11 mutations in a 893-bp segment with maximal distances between any two variants of only 5 mutations (0.6%), including a characteristic 16-bp insertion/deletion. These HPV-35 variants formed several phylogenetic clusters rather than two- or three-branched trees as HPV-16, 18, and 31. An HPV-35 variant typical for American Indians was not identifiable. Our research suggests type specific patterns of evolution and spread of HPV-16, 18, 31, and 35 both before and after the worldwide migrations of the last four centuries. The high prevalence of highly carcinogenic HPV-16 AA variants, and the extensive diversity of HPV-18, 31, and 35 variants with unknown pathogenic properties raise the possibility that HPV intra-type variation contributes to the high cervical cancer burden in Mexico

Evaluación de la diversidad de Manihot esculenta mediante huellas genéticas para reducir la redundancia en las colecciones de germoplasma

El análisis de las huellas genéticas es una poderosa herramienta para comprender la diversidad genética de las poblaciones, ya que cada individuo tiene fragmentos únicos de secuencias de ADN que lo identifican. Una adecuada comprensión de la diversidad de las poblaciones de yuca proporciona información relevante para la toma de decisiones en el cultivo, su conservación y mejoramiento. Evaluamos la diversidad genética de la yuca utilizando el ensayo tipo SNP-Chip de Fluidigm con el objetivo de explorar las relaciones genéticas entre individuos. Esto permitirá reducir la redundancia en los bancos de germoplasma y así mismo el tiempo y costo de mantenimiento de las colecciones

Algunas evidencias de aplicación

Libro temático especializadoLa sustentabilidad también debe aplicarse al sistema de producción, buscando impulsar transformaciones graduales de los estilos y modelos productivos tradicionales a unas de mayor eficiencia. Y donde se incorpore la dimensión ambiental y geográfico-espacial, para crear estructuras productivas más progresivas y equitativas en las sociedades. Todo esto, como alternativa para revertir las tendencias de escasez y agotamiento de los recursos naturales, así como de los desequilibrios globales, cuyos costos permean todos los tejidos humanos. De esta manera, la “sustentabilidad productiva” se concibe como la generación de bienes y servicios con ciertos estándares de calidad, bajo un esquema de eficiencia, rendimiento y de organización inclusiva e integrada, con baja presión al ambiente y uso racional de los recursos, garantizando la estadía y permanencia de los insumos y materiales en el tiempo. Desde esta perspectiva, la producción sustentable y el crecimiento de largo plazo pueden ser explicados por la capacidad que tienen las economías para generar e incorporar conocimientos y tecnologías. De ahí que, la educación y las cualificaciones del capital humano, los cambios en la organización de la producción y la calidad institucional, sean elementos nodales para avanzar en la consolidación de este ambiente productivo

Timber production and biodiversity conservation in Nothofagus forests of Tierra del Fuego and southern Patagonia.

Las propuestas de manejo forestal para Tierra del Fuego y Patagonia Sur se basan en el manejo silvopastoril para Nothofagus antarctica (ñire), y cortas de protección y raleo para Nothofagus pumilio (lenga). Ambas propuestas producen impactos sobre los componentes bióticos y abióticos del bosque original. El objetivo de este capítulo es analizar las propuestas de manejo, planteando alternativas que prioricen el equilibrio entre producción y conservación a partir de las investigaciones actuales. Se analizan diferentes escalas en la planificación del manejo forestal y estrategias de conservación (macro, meso y micro-escala), describiendo ventajas y costos incrementales de su aplicación. En particular, se describe la aplicación de la retención variable como técnica complementaria de las cortas de protección, para minimizar los impactos de la cosecha a escala de rodal sobre las variables abióticas y bióticas. Asimismo, se analiza la regeneración natural como la variable de mayor importancia en los monitoreos postcosecha, junto con los factores limitantes del ciclo y la posterior dinámica en parcelas de investigación a largo plazo, así como los resultados de ensayos de raleos y podas comerciales. Finalmente, se describen las carencias en el conocimiento científico y técnico desarrollado hasta el presente, a fin de mejorar la implementación del manejo forestal actual. A partir de este análisis se proponen diez desafíos a tener en cuenta para la próxima década. Summary: Forest management for Tierra del Fuego and southern Patagonia are based mainly on silvopastoral use of Nothofagus antarctica (ñire), and shelterwood cuts and thinnings for Nothofagus pumilio (lenga). Both proposals had impacts over biotic and abiotic components of the original forests. The objective of this chapter was to analyze these management practices by introducing a concept of equilibrium between timber production and conservation. Different planning scales (macro-, meso- and micro-scale) were analyzed for forest management and conservation including advantages and implementation. In particular, variable retention implementation was described as complementary technique of the shelterwood cuts, where harvesting impacts over abiotic and biotic scale at stand level were minimized. Natural regeneration, as one of the most important variable in the post-harvesting monitoring, also was analyzed. Limiting factors in the whole reproductive cycle, their natural dynamics in long-term research plots, commercial thinnings and prunings were described. Finally, it was identified the main actual scientific and technical knowledge gaps in order to improve the future research and implementation in the current forest management.Forest management for Tierra del Fuego and southern Patagonia are based mainly on silvopastoral use of Nothofagus antarctica (ñire), and shelterwood cuts and thinnings for Nothofagus pumilio (lenga). Both proposals had impacts over biotic and abiotic components of the original forests. The objective of this chapter was to analyze these management practices by introducing a concept of equilibrium between timber production and conservation. Different planning scales (macro-, meso- and micro-scale) were analyzed for forest management and conservation including advantages and implementation. In particular, variable retention implementation was described as complementary technique of the shelterwood cuts, where harvesting impacts over abiotic and biotic scale at stand level were minimized. Natural regeneration, as one of the most important variable in the post-harvesting monitoring, also was analyzed. Limiting factors in the whole reproductive cycle, their natural dynamics in long-term research plots, commercial thinnings and prunings were described. Finally, it was identified the main actual scientific and technical knowledge gaps in order to improve the future research and implementation in the current forest management.EEA Santa CruzFil: Martínez Pastur, Guillermo José. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Austral de Investigaciones Científicas (CADIC); Argentina.Fil: Peri, Pablo Luis. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Santa Cruz; Argentina.Fil: Peri, Pablo Luis. Universidad Nacional de la Patagonia Austral; Argentina.Fil: Peri, Pablo Luis. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina.Fil: Lencinas, María Vanessa. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Austral de Investigaciones Científicas (CADIC); Argentina.Fil: Cellini, Juan Manuel. Universidad Nacional de la Plata. Facultad de Ciencias Naturales y Museo. Laboratorio de Investigaciones en Maderas; Argentina.Fil: Barrera, Marcelo Daniel. Universidad Nacional de la Plata. Facultad de Ciencias Naturales y Museo. Laboratorio de Investigaciones en Maderas; Argentina.Fil: Soler Esteban, Rosina Matilde. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Austral de Investigaciones Científicas (CADIC); Argentina.Fil: Ivancich, Horacio Simón. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Austral de Investigaciones Científicas (CADIC); Argentina.Fil: Mestre, Luciana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Austral de Investigaciones Científicas (CADIC); Argentina.Fil: Moretto, Alicia Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Austral de Investigaciones Científicas; Argentina.Fil: Anderson, Christopher B. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina.Fil: Anderson, Christopher B. Universidad Nacional de Tierra del Fuego; Argentina.Fil: Pulido, Fernando. Universidad de Extremadura. EUIT Forestal. Forest Research Group; España