Enzymes are in high demand for very diverse biotechnological applications. However, natural biocatalysts often need to be engineered for fine-tuning their properties towards the end applications, such as the activity, selectivity, stability to temperature or co-solvents, and solubility. Computational methods are increasingly used in this task, providing predictions that narrow down the space of possible mutations significantly and can enormously reduce the experimental burden. Many computational tools are available as web-based platforms, making them accessible to non-expert users. These platforms are typically user-friendly, contain walk-throughs, and do not require deep expertise and installations. Here we describe some of the most recent outstanding web-tools for enzyme engineering and formulate future perspectives in this field.

Soils provide important regulating ecosystem services and have crucial implications for human well-being and environmental conservation. However, soil degradation and particularly soil erosion jeopardize the maintenance and existence of these services. This study explores the spatio-temporal relationships of soil erosion to understand the distribution patterns of sediment retention services in mainland Portugal. Based on Corine Land Cover maps from 1990 to 2018, the InVEST Sediment Delivery Ratio (SDR) model was used to evaluate the influence of sediment dynamics for soil and water conservation. Spatial differences in the sediment retention levels were observed within the NUTS III boundaries, showing which areas are more vulnerable to soil erosion processes. Results indicated that the Region of Leiria, Douro and the coastal regions have decreased importantly sediment retention capacity over the years. However, in most of the territory (77.52%) changes in sediment retention were little or not important (i.e., less than 5%). The statistical validation of the model proved the consistency of the results, highlighting the usefulness of this methodology to analyse the state of soil erosion in the country. These findings can be relevant to support strategies for more efficient land use planning regarding soil erosion mitigation practices.

In this paper we investigate in different scenarios the feasibility of using massive multiple-input multiple-output (mMIMO) with Reconfigurable Intelligent Surfaces (RISs) to boost the throughput and coverage performances with high energy efficiency, considering sub-6 GHz, mmWave and THz bands. With that objective, a centralized radio access network (C-RAN) suitable for beyond fifth generation (B5G) systems is considered, where we integrate the base stations (BSs) with multiple RISs and devices or user equipment. RISs with a large number of quasi-passive reflecting elements constitute a low-cost approach capable of shaping the radio wave propagation and improve wireless connectivity. We consider a scenario where multiple RISs are combined with mMIMO in the uplink in order to provide connectivity to Smart City (thousands of active low power devices) wirelessly, in the 3.6GHz and 28GHz bands. We also address a scenario where RISs are adopted with mMIMO in the downlink so as to offer connectivity to a Stadium with Pitch, (thousands of active user equipment) in the 28GHz band. Finally, we have also studied the connectivity at 100GHz of a Factory where several RIS panels replacing most of the BSs equipped with mMIMO assure improved throughput and coverage. We concluded that RISs are capable of improving the performance in any of these analyzed scenarios at the different frequency bands, justifying that they are a key enabling technology for 6G.

A single farmed fish species assimilates about 20% of the nutrients in the supplied diet. This study evaluated if the culture of complementary ecological-function species can recover nutrients dispersed into the water and transform them into high-valued biomass. A completely randomized experiment was designed with three treatments and four replications of each production system: monoculture of lambari (Astyanax lacustris); integrated aquaculture of lambari and Amazon river prawn (Macrobrachium amazonicum); and integrated aquaculture of lambari, Amazon river prawn, and curimbatá (Prochilodus lineatus). Fingerlings of lambari (0.8 ± 0.8 g) were stocked in twelve earthen-ponds (0.015 ha) at the density of 50 fish m^-2. Eight ponds, were stocked with juveniles of Amazon river prawn (1.1 ± 0.2 g) at the density of 25 prawn m⁻². Four of these eight ponds were stocked with curimbatá fingerlings (0.2 ± 0.1 g) at a density of 13 fish m^-². Only lambari was fed twice a day with an extruded commercial diet. The experiment lasted 60 days when lambari attained commercial size. The inclusion of prawn increased the total species yield from 1.8 to 2.4 t ha^-1 cycle^-1 and reduced the feed conversion ratio (FCR) from 2.5 to 1.8. The inclusion of prawn and curimbatá increased the total yield to 3.2 t ha^-1 cycle^-1 and reduced the FCR to 1.4. Therefore, the integrated culture of lambari, prawn, and curimbatá improves the use of space, water, feed, and benthic species can recover the large quantity of nutrients accumulated in the bottom of lambari pond production, converting them into high-nutritional and monetary-valued biomass.

Locally advanced rectal cancer (LARC) has traditionally been treated with trimodality therapy consisting of neoadjuvant radiation +/- chemotherapy, surgery and adjuvant chemotherapy. There is currently a clinical need for biomarkers to predict treatment response and outcomes, especially during neoadjuvant therapy. Liquid biopsy in the form of circulating tumour cells (CTCs) and circulating nucleic acids in particular microRNAs (miRNA) are novel, highly stable and clinically relevant regulators of disease. We studied a prospective cohort of 52 patients with LARC, and obtained samples at baseline, during treatment, and post-treatment. We enumerated CTCs during chemoradiation at these three time-points, using the IsofluxTM CTC Isolation and detection platform. We then subjected the isolated CTCs to miRNA expression analyses, using a panel of 106 miRNA candidates. We identified CTCs in 73% of patients at baseline, and numbers fell during treatment and miRNA expression profiles also changed during treatment. Between baseline and during treatment (week 3) time-points three microRNAs (hsa-miR-95, hsa-miR-10a and hsa-miR-16-1*) were highly differentially expressed. Importantly, hsa-miR-19b-3p and hsa-miR-483-5p were found to correlate with good response to treatment. The latter (hsa-miR-483-5p) was also found to be differentially expressed between good responders and poor responders. They represent potential predictive biomarkers and a potential miRNA-based treatment strategy. In this study, we demonstrate that CTCs are present and can be isolated in the non-metastatic early stage cancer setting, and their associated miRNA profiles can potentially be utilized to predict treatment response.