Efficient Base-Catalyzed Kemp Elimination in an Engineered Ancestral Enzyme

The routine generation of enzymes with completely new active sites is a major unsolved problem in protein engineering. Advances in this field have thus far been modest, perhaps due, at least in part, to the widespread use of modern natural proteins as scaffolds for de novo engineering. Most modern proteins are highly evolved and specialized and, consequently, difficult to repurpose for completely new functionalities. Conceivably, resurrected ancestral proteins with the biophysical properties that promote evolvability, such as high stability and conformational diversity, could provide better scaffolds for de novo enzyme generation. Kemp elimination, a non-natural reaction that provides a simple model of proton abstraction from carbon, has been extensively used as a benchmark in de novo enzyme engineering. Here, we present an engineered ancestral beta-lactamase with a new active site that is capable of efficiently catalyzing Kemp elimination. The engineering of our Kemp eliminase involved minimalist design based on a single function-generating mutation, inclusion of an extra polypeptide segment at a position close to the de novo active site, and sharply focused, low-throughput library screening. Nevertheless, its catalytic parameters (k(cat)/K-M similar to 2.10(5) M-1 s(-1), k(cat)similar to 635 s(-1)) compare favorably with the average modern natural enzyme and match the best proton-abstraction de novo Kemp eliminases that are reported in the literature. The general implications of our results for de novo enzyme engineering are discussed.Human Frontier Science Program RGP0041/2017Spanish Government RTI-2018-097142-B100 EQC2019-006403-PFEDER/Junta de Andalucia-Consejeria de Economia y Conocimiento E.FQM.113.UGR1

FAST-SE: An Esb Based Framework for SLA Trading

SLA driven service transaction has been identified as a key challenge to take advantage of a SOA. FAST System provides a software framework for the automated creation of SLAs. in particular it have been developed as an extension to the ESB (Enterprise Service Bus) paradigm to create a transparent SLA management layer that drives any service invocation. Our framework has been successfully applied in two different scenarios and provides an extensible architecture to address new domains

WS-Governance Tooling: SOA Governance Policies analysis and authoring

Governance is a capital issue in current Service Oriented Arcuitectures, and governance policies are at its base. The governance policies definition must be supported by proper languages and tools, allowing for comfortable and collaborative editing, consistency checking and the evaluation policy meeting. In this paper we present a policy analizer for WS-Governance (a governance policy definition language created by authors, described in [1]) together with an online editor and test suite with classical examples of WS-Governance Documents for consistency validation. Both the test model and the analysis tool prove the suitability of WS-Governance to define SOA governance policiesComisión Interministerial de Ciencia y Tecnología (CICYT) SETI (TIN2009-07366)Junta de Andalucía TIC-2533Junta de Andalucía TIC-590

Automated analysis of feature models: Quo vadis?

Feature models have been used since the 90's to describe software product lines as a way of reusing common parts in a family of software systems. In 2010, a systematic literature review was published summarizing the advances and settling the basis of the area of Automated Analysis of Feature Models (AAFM). From then on, different studies have applied the AAFM in different domains. In this paper, we provide an overview of the evolution of this field since 2010 by performing a systematic mapping study considering 423 primary sources. We found six different variability facets where the AAFM is being applied that define the tendencies: product configuration and derivation; testing and evolution; reverse engineering; multi-model variability-analysis; variability modelling and variability-intensive systems. We also confirmed that there is a lack of industrial evidence in most of the cases. Finally, we present where and when the papers have been published and who are the authors and institutions that are contributing to the field. We observed that the maturity is proven by the increment in the number of journals published along the years as well as the diversity of conferences and workshops where papers are published. We also suggest some synergies with other areas such as cloud or mobile computing among others that can motivate further research in the future.Ministerio de Economía y Competitividad TIN2015-70560-RJunta de Andalucía TIC-186

Flapless implant surgery: a review of the literature and 3 case reports

Since the 1970s, modern Implantology is based on a concept of surgery with flap elevation. Gradually, several clinical trials demonstrated that a mid-crestal incision gives similar success rates compared to those obtained using the classical protocol. However, over the past decade in medicine it has been established the concept of minimally invasive surgery, consisting in taking advantage of advancements experienced in diagnostic techniques and specific surgical instruments, to perform surgical procedures infringing as less damage as possible to the patient The present work aims to produce a thorough review of the literature published on the field of Implantology with flapless surgery, to determine the current scientific evidence of the technique, along with illustrating the results with different clinical cases. After presenting the clinical cases, and the review of literature, we can say that flapless surgeries should be restricted to well-selected cases in which a proper clinical and radiological planning has been made. Patients treated with anticoagulant drugs or medically compromised equally can get benefitted by this minimal invasion technique

El foro romano de Iuliobriga (Cantabria): nuevas investigaciones arqueológicas

This article offers a general interpretation of Iuliobriga’s forum, emphasizing its characteristics and peculiarities from the information obtained in the latest archaeological research. It focuses on the results of an excavation made in 2004 inside the church of Santa María in Retortillo. This archaeological dig has allowed us to broaden the view we had of the forum’s square, thanks to the discovery of a new functional space inside it. The archaeological research has also revealed information about the origin of the habitat in Iuliobriga and permits the forum to be dated with certainty in the Flavian period.Se ofrece una interpretación general del foro de Iuliobriga, destacando sus características y peculiaridades tras la información obtenida en las últimas investigaciones arqueológicas. En especial, se presentan los resultados de una excavación realizada en 2004 en el interior de la iglesia de Santa María de Retortillo, la cual ha permitido ampliar la visión que se tenía de la plaza porticada del foro, con el descubrimiento de un nuevo espacio funcional dentro de éste. La misma intervención arqueológica ha aportado datos sobre el origen del hábitat en Iuliobriga y sobre la datación flavia del conjunto monumental del foro

Hidratació i rendiment: pautes per a una elusió efectiva de la deshidratació per exercici

El rendiment de les capacitats físiques i mentals durant la pràctica esportiva o la competició, es troba disminuït sota condicions de deshidratació. La pèrdua de pes es deu a la pèrdua d’aigua (suor i respiració) i quan aquesta és important, afecta de manera decisiva els sistemes nerviós, cardiovascular, termoregulador, endocrí i/o metabòlic, i pot provocar fins i tot la mort. Per eludir aquests efectes negatius de la deshidratació així com la disminució del rendiment, l’atleta haurà de beure prou quantitat de líquid abans, durant, i després de la competició o la pràctica d’exercici. Idealment, la beguda serà una solució composta d’aigua, electròlits i carbohidrats, en quantitat adequada per garantir, d’una banda, un rendiment òptim durant la competició i, d’una altra, reposar eficaçment i completament les pèrdues hídriques, electrolítiques i energètiques. La composició i el volum de la beguda tindrà en compte els factors extrínsecs (temperatura, humitat, altitud, vent, etc.) que envolten la competició, els factors intrínsecs de l’atleta (nivell de deshidratació, problemes gastrointestinals, tipus de competició) i les pròpies característiques de la beguda (gust, temperatura, composició). En termes generals, els atletes no acostumen a beure, durant la pràctica esportiva, el volum necessari per reemplaçar la pèrdua de fluids i electròlits, cosa que fa que calgui establir pautes perquè ho facin. En aquest treball es presenten aquestes pautes i la base fisiològica que les sosté

Hidratación y rendimiento: pautas para una elusión efectiva de la deshidratación por ejercicio

El rendimiento de las capacidades físicas y mental durante la práctica deportiva o la competición, está mermado bajo condiciones de deshidratación. La pérdida de peso se debe a la pérdida agua (sudor y respiración) y cuando ésta es importante, afecta de manera decisiva a los sistemas nervioso, cardiovascular, termorregulador, endocrino y/o metabólico, pudiendo provocar incluso la muerte. Para eludir estos efectos negativos de la deshidratación así como la disminución del rendimiento, el atleta deberá beber suficiente cantidad de líquido antes, durante y después de la competición o la práctica de ejercicio. Idealmente, la bebida será una solución compuesta de agua, electrolitos y carbohidratos en cantidad adecuada para garantizar, por un lado, un óptimo rendimiento durante la competición y, por otro, reponer eficaz y completamente las pérdidas hídricas, electrolíticas y energéticas. La composición y el volumen de la bebida tendrá en cuenta los factores extrínsecos (temperatura, humedad, altitud, viento, etc.) que rodean la competición, los factores intrínsecos del atleta (nivel de deshidratación, problemas gastrointestinales, tipo de competición) y las propias características de la bebida (sabor, temperatura, composición). En términos generales, los atletas no suelen beber, durante la práctica deportiva, el volumen necesario para reemplazar la pérdida de fluidos y electrolitos, lo que hace necesario establecer pautas para que lo hagan. En el presente trabajo se presentan esas pautas y la base fisiológica que las sustentan

Infecciones orofaciales de origen odontogénico

La naturaleza polimicrobiana de las infecciones odontógenas así como la heterogeneidad de los cuadros clínicos asociados son consecuencia de la diversidad de la microbiota bucal y de la complejidad anatómica y funcional de la cavidad oral. Así mismo, estos procesos pueden dar lugar a múltiples complicaciones que pueden limitarse a afecciones locales o derivar hasta compromisos sistémicos. En el tratamiento farmacológico de estas infecciones es crucial la elección del antibiótico y la pauta posológica más eficaces. La farmacodinamia proporciona los parámetros que hacen posible valorar como varía la actividad de los antibióticos en función del tiempo. Como norma general, en el manejo inicial de las infecciones orofaciales en el adulto, incluidas las infecciones odontógenas, se utilizará amoxicilina/ac. Clavulánico en dosis de 875 mg de amoxicilina y 125 mg de ac. clavulánico pautado cada 8 horas. El cumplimiento terapéutico es clave para evitar la aparición de resistencias, por lo que se ha de maximizar la aceptación por parte de los pacientes. En este sentido se ha demostrado que la nueva presentación de 2000/125 mg de Augmentine Plus pautado cada 12 horas tanto en profilaxis como tratamiento disminuye significativamente la tasa de complicaciones infecciosas derivada de la extracción del tercer molar.The polymicrobial nature of the odontogenic infections as well as the variety of associated conditions are a consequence of the diversity of the buccal microbiota and the anatomical and functional complexity of the oral cavity. In addition to this, all these processes can give way to multiple complications which range from the local to the systemic level. The appropriate choice of antibiotic and posology is crucial in the successful management of these infections. Pharmacodynamics provides those parameters that make it possible to assess how antibiotics activity varies in time. As a general rule, the first step in the initial management of orofacial infections in adults, included odontogenic infections, will be the administration of 875 mg of amoxicillin and 125 mg of clavulanic every 8 hours. Therapeutic compliance is paramount to avoid resistance, therefore patient's acceptance must be sought. In this sense, it has been proved that Augmentine Plus (2000/125) every twelve hours both as profylaxis and as treatment significantly decreases the rate of infective complications associated to extraction of the third molar

Protection of Catalytic Cofactors by Polypeptides as a Driver for the Emergence of Primordial Enzymes

Enzymes catalyze the chemical reactions of life. For nearly half of known enzymes, catalysis requires the binding of small molecules known as cofactors. Polypeptide-cofactor complexes likely formed at a primordial stage and became starting points for the evolution of many efficient enzymes. Yet, evolution has no foresight so the driver for the primordial complex formation is unknown. Here, we use a resurrected ancestral TIM-barrel protein to identify one potential driver. Heme binding at a flexible region of the ancestral structure yields a peroxidation catalyst with enhanced efficiency when compared to free heme. This enhancement, however, does not arise from proteinmediated promotion of catalysis. Rather, it reflects the protection of bound heme from common degradation processes and a resulting longer lifetime and higher effective concentration for the catalyst. Protection of catalytic cofactors by polypeptides emerges as a general mechanism to enhance catalysis and may have plausibly benefited primordial polypeptide-cofactor associations.Human Frontier Science Program grant RGP0041/2017National Science Foundation grant 2032315Department of Defense grant MURI W911NF-16-1-0372National Institutes of Health grant R01AR069137Spanish Ministry of Science and Innovation/ FEDER Funds grant PID2021-124534OB-100Grant PID2020-116261GB-I0