Efficient extraction of the RuBisCO enzyme from spinach leaves using aqueous solutions of biocompatible ionic liquids

Ribulose-1,5-biphosphate carboxylase/oxygenase (RuBisCO) is the most abundant protein on the planet, being present in plants, algae and various species of bacteria, with application in the pharmaceutical, chemical, cosmetic and food industries. However, current extraction methods of RuBisCO do not allow high yields of extraction. Therefore, the development of an efficient and selective RuBisCOs’ extraction method is required. In this work, aqueous solutions of biocompatible ionic liquids (ILs), i.e., ILs derived from choline and analogues of glycine-betaine, were applied in the RuBisCO’s extraction from spinach leaves. Three commercial imidazolium-based ILs were also investigated for comparison purposes. To optimize RuBisCO’s extraction conditions, response surface methodology was applied. Under optimum extraction conditions, extraction yields of 10.92 and 10.57 mg of RuBisCO/g of biomass were obtained with the ILs cholinium acetate ([Ch][Ac]) and cholinium chloride ([Ch]Cl), respectively. Circular dichroism (CD) spectroscopy results show that the secondary structure of RuBisCO is better preserved in the IL solutions when compared to the commonly used extraction solvent. The obtained results indicate that cholinium-based ILs are a promising and viable alternative for the extraction of RuBisCO from vegetable biomass.publishe

Integrated platform resorting to ionic liquids comprising the extraction, purification and preservation of DNA

The large-scale production of therapeutically targeted-deoxyribonucleic acid (DNA) has passed through several challenges, postponing the tangible implementation of an effective, economic and sustainable manufacturing system. Such challenges comprise the need to develop an integrative downstream process able to extract, purify and long-term preserve DNA, whilst reducing the risk of degradation by endonucleases that would compromise their effectiveness as therapeutic products. In this work, three-phase partitioning (TPP) systems formed by the application of aqueous biphasic systems (ABS) composed of several biocompatible cholinium-based ionic liquids (ILs), are proposed for the separation of double stranded DNA (dsDNA) from the endonuclease deoxyribonuclease I (DNase I). By taking advantage of the tailor-made properties of ILs, dsDNA can be completely extracted to the IL-rich phase, whereas DNase I is precipitated at the ABS interphase. The ABS/TPP formed by IL cholinium glycolate ([N 111(2OH) ][Gly]) fulfills the aim of this work, i.e. at ensuring the technical viability of IL-based ABS/ TPP for the “one-pot” extraction, purification and long-term preservation of dsDNA. The results reveal the potential of this system to be applied in the bioprocessing of DNA, particularly relevant when envisioning DNA- based therapeutic products.publishe

Sustainable liquids support as a media for biocatalytic reactions using aqueous biphasic systems

Over the last few years a significant interest in the application of biocatalyst in several industries as an alternative to the commonly used chemical biocatalyst has emerged, since enzymes, such as laccase, allow for an improved, sustainable and biodegradable catalytic process partially due to their high specificity to its substrates. Nevertheless, its application can still be considered a costly process thus to overcome this shortcoming emerges the need for the enzyme reutilization employed in the catalytic reaction 1. One of the alternatives is to implement aqueous biphasic systems (ABS), which are a biocompatible liquid-liquid extraction as a liquid support. Moreover, the introduction of ionic liquids (ILs) in ABS have shown promising results in the system overall performance and could potentially allow for an improved biocatalyst performance. Therefore, our goal was to form an ABS composed of cholinium-based ILs and polypropylene glycol (PPG 400) as novel liquid supports for enzymes, using laccase as models in order to evaluated the degradation of the textile dye Remazol Brilliant Blue R (RBBR) as well as the capacity to reutilized the biocatalyst for further applications.publishe

High resolution forecasts of wind in rough orography and in coastal conditions

The increase of the share of wind energy in total electrical power requires the development of reliable forecasts of wind power at different time scales, from minutes to days, on a regional and national basis. That development is multidisciplinary, involving meteorological and engineering components. In the present stage, though, it is thought that for time scales beyond 3 hours, the largest improvements are expected from the meteorological side, if meteorological forecasts are able to deal with mesoscale flow. In this paper three case studies of mesoscale modeling are analyzed and compared against wind observations in sites selected for their high potential for wind energy. Two cases deal with flow in the Islands of Madeira and San Jorge (Azores), characterized by rough orography, where measurements were made at the top of elevated plateaus. In these two cases, the large scale flow is obtained from ECMWF analysis and the mesoscale flow is computed, for one summer and one winter month, with two mesoscale models at resolutions between 6 and 0.5 km. In the third case, one full year of simulationis Continental Portugal is performed at resolutions between 27 and 3km, using boundary conditions from the NCEP/NCAR reanalysis and the MM5 model. Results indicate the interest of high horizontal resolution for wind forecasts in complex terrain but also some difficulties in the representation of boundary layer effects in conditions of large thermal forcing.

Robustness Diagram: a bridge between business modelling and system design

Use case driven development has proven being a good approach for capturing problem semantics in an orderly, structured description. However, it specifies an abstraction beyond practicity to guide system design process. Robustness diagrams are a simple solution for drafting a more formal description for business modelling. This very simplicity may, however, detract its value, falling short of capturing the rich business semantics. Stereotyping is the essence behind its robustness diagram mechanics. Symbols convey the abstraction necessary to catch the model semantics. Increasing the number of stereotypes we can achieve a closer match from model to design. Rules carefully stated for robustness diagram can help to translate highleveI information into well-behaved and predictable symbolic descriptions. This enhancement to robustness diagram helps to patch the gap between abstract model and project into a paved continuum. It has been used to train programmers to extract working and consistent systems out of use case specifications

Simulated climate change, but not predation risk, accelerates Aedes aegypti emergence in a microcosm experiment in western Amazonia

Climate change affects individual life-history characteristics and species interactions, including predator-prey interactions. While effects of warming on Aedes aegypti adults are well known, clarity the interactive effects of climate change (temperature and CO2 concentration) and predation risk on the larval stage remains unexplored. In this study, we performed a microcosm experiment simulating temperature and CO2 changes in Manaus, Amazonas, Brazil, for the year 2100. Simulated climate change scenarios (SCCS) were in accordance with the Fourth Assessment Report of Intergovernmental Panel on Climate Change (IPCC). Used SCCS were: Control (real-time current conditions in Manaus: average temperature is ~25.76°C ± 0.71°C and ~477.26 ± 9.38 parts per million by volume (ppmv) CO2); Light: increase of ~1, 7°C and ~218 ppmv CO2; Intermediate: increase of ~2.4°C and ~446 ppmv CO2; and Extreme: increase of ~4.5°C and ~861 ppmv CO2, all increases were relative to a Control SCCS. Light, Intermediate and Extreme SCCS reproduced, respectively, the B1, A1B, and A2 climatic scenarios predicted by IPCC (2007). We analyzed Aedes aegypti larval survivorship and adult emergence pattern with a factorial design combining predation risk (control and predator presence-Toxorhynchites haemorrhoidalis larvae) and SCCS. Neither SCCS nor predation risk affected Aedes aegypti larval survivorship, but adult emergence pattern was affected by SCCS. Accordingly, our results did not indicate interactive effects of SCCS and predation risk on larval survivorship and emergence pattern of Aedes aegypti reared in SCCS in western Amazonia. Aedes aegypti is resistant to SCCS conditions tested, mainly due to high larval survivorship, even under Extreme SCCS, and warmer scenarios increase adult Aedes aegypti emergence. Considering that Aedes aegypti is a health problem in western Amazonia, an implication of our findings is that the use of predation cues as biocontrol strategies will not provide a viable means of controlling the accelerated adult emergence expected under the IPCC climatic scenarios

Total fishmeal replacement by defatted Tenebrio molitor larvae meal induces alterations in intermediary metabolism of European sea bass (Dicentrarchus labrax)

The replacement of fishmeal (FM) by insect meal (IM) in aquafeed formulation has been thoroughly studied lately, but little is known about their impact on nutrient metabolism of fish. This study evaluated the impact not only of partial but also total FM replacement by IM on intermediary metabolism of European sea bass (Dicentrarchus labrax). A fishmeal-based diet was used as a control (CTRL) and two other diets were formulated to include 20% and 40% of defatted Tenebrio molitor larvae meal (dTM), replacing 50% (TM50) and 100% (TM100) of fishmeal (FM), respectively. After a 16-week feeding trial, a multidisciplinary approach including assessment of histological, biochemical, molecular, and enzymatic parameters was adopted to investigate hepatic and plasmatic responses to the different dietary formulations. The results obtained demonstrated that dTM can be successfully used to replace 50% of FM in diets for European sea bass, without adversely affecting liver health or intermediary metabolism of nutrients. As for TM100, although no signs of steatosis were observed in the liver, the activity of glycolytic and lipogenic genes and enzymes increased when compared to CTRL diet (P < 0.05), resulting in higher levels of plasmatic non-esterified fatty acids and triacylglycerides (P < 0.05), which in the long-term may compromise fish health, thus precluding such a high degree of substitution for use in practical diets for European sea bass.Agencia Estatal de Investigación | Ref. PID2019-103969RB-C31Xunta de Galicia | Ref. ED431B 2022/01Fundação para a Ciência e a TecnologiaUniversidade de Vigo/CISU

Smart systems for monitoring buildings - an IoT application

Life in society has initiated a search for comfort and security in social centers. This search generated revolutions within the knowledge about the technologies involved, making the environments automated and integrated. Along with this increase, ecological concerns have also arisen, which have been involved since the design of intelligent buildings, remaining through the years of their use. Based on these two pillars, the present study aims to monitor three central systems inside the apartments of the Apolo Building (Bragan¸cacity, Portugal). The electrical energy consumption, water flow, and waste disposal systems are integrated through a single database. The data is sent remotely via WiFi through the microcontroller. For better visualization and analytics of the data, a web application is also developed, which allows for real-time monitoring. The obtained results demonstrate to the consumer his behavior regarding household expenses. The idea of showing the consumer their expenditure is to create an ecological awareness. Through the data collected and the environmental alternatives found, it is possible to observe whether there was a behavior change when receiving this data, either in the short or long term.The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support through national funds FCT/MCTES (PIDDAC) to CeDRI (UIDB/05757/2020 and UIDP/05757/2020) and SusTEC (LA/P/0007/2021). Thadeu Brito was supported by FCT PhD grant SFRH/BD/08598 /2020 and João Braun received the support of a fellowship from ”la Caixa” Foundation (ID 100010434) with code LCF/BQ/DI20/11780028.info:eu-repo/semantics/acceptedVersio