Publications by authors named "Ricardo J Silva"

A biodegradable polymer packaging system for 'Benitaka' table grapes ( L.) was developed to inhibit the development of gray mold during refrigerated storage. The system consisted of packages and sachets containing NaSO to release sulfur dioxide (SO), both produced with biodegradable films of starch, glycerol, and poly (adipate co-butylene terephthalate) (PBAT) produced via blown extrusion.

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Staphylococcus aureus is increasingly recognized as a facultative intracellular pathogen, although the significance and pervasiveness of its intracellular lifestyle remain controversial. Here, we applied fluorescence microscopy-based infection assays and automated image analysis to profile the interaction of 191 S. aureus isolates from patients with bone/joint infections, bacteremia, and infective endocarditis, with four host cell types, at five times post-infection.

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Modulation of the host cell cycle has emerged as a common theme among the pathways regulated by bacterial pathogens, arguably to promote host cell colonization. However, in most cases the exact benefit ensuing from such interference to the infection process remains unclear. Previously, we have shown that actively induces G/M arrest of host cells, and that infection is severely inhibited in cells arrested in G.

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MicroRNAs (miRNAs) are increasingly recognized for their role in infection by bacterial pathogens, although the effect of each individual miRNA remains largely unknown. Here, we used a comparative genome-wide microscopy-based functional screening approach to identify miRNAs controlling infection by two bacterial pathogens-Salmonella enterica serovar Typhimurium and Shigella flexneri. Despite the similarities between these pathogens, we found infections to be controlled by largely non-overlapping subsets of miRNAs, seemingly reflecting different requirements prompted by their distinct intracellular lifestyles.

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The indirect identification of doping in sports can be performed by assessing athletes' hematological perturbations from the analysis of blood collected on different occasions. Because prosecution for doping based on this information requires expensive and time-consuming interpretation of blood analysis results by various expert hematologists, mathematical data screening is performed to decide which cases should be forwarded to hematologists. The current Bayesian and univariate screening of data does not process the multivariate trends of blood parameters or take the time interval between samplings into account.

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MicroRNAs (miRNAs) are small non-coding RNAs that post-transcriptionally modulate gene expression and orchestrate a wide range of biological and pathological processes. The use of high-throughput screening technologies, in particular microscopy-based screenings (also known as high-content screenings), coupled with genome-wide libraries for modulation of miRNA levels, allow for comprehensive functional analysis of each member of the miRNome in different phenotypic cell-based assays. The wealth of information obtained from such screenings spans across various fields of research, including cancer, cardiovascular, cell reprogramming, and infection biology.

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A hybrid simulation model (macro-molecular dynamics and Monte Carlo method) is proposed to reproduce neurosecretion and exocytosis. A theory has been developed for vesicular dynamics based on quasi-static electric interactions and a simple transition-state model for the vesicular fusion. Under the non-equilibrium electric conditions in an electrolytic fluid, it is considered that the motion of each synaptic vesicle is influenced by electrostatic forces exerted by the membranes of the synaptic bouton, other vesicles, the intracellular and intravesicular fluids, and external elements to the neuron.

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MicroRNAs play an important role in the interplay between bacterial pathogens and host cells, participating as host defense mechanisms, as well as exploited by bacteria to subvert host cellular functions. Here, we show that microRNAs modulate infection by Shigella flexneri, a major causative agent of bacillary dysentery in humans. Specifically, we characterize the dual regulatory role of miR-29b-2-5p during infection, showing that this microRNA strongly favors Shigella infection by promoting both bacterial binding to host cells and intracellular replication.

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The conservation and sustainable management of Annona coriacea requires knowledge of its floral and reproductive biology, and of its main pollinators and their life cycles. In this work, we analyzed these aspects in detail. Floral biology was assessed by observing flowers from the beginning of anthesis to senescence.

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The use of human mesenchymal stem cells (hMSC) in clinical applications has been increasing over the last decade. However, to be applied in a clinical setting hMSC need to comply with specific requirements in terms of identity, potency and purity. This study reports the improvement of established tangential flow filtration (TFF)-based washing strategies, further increasing hMSC purity, using negative mode expanded bed adsorption (EBA) chromatography with a new multimodal prototype matrix based on core-shell bead technology.

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This commentary is intended to find possible explanations for the low impact of computational modeling on pain research. We discuss the main strategies that have been used in building computational models for the study of pain. The analysis suggests that traditional models lack biological plausibility at some levels, they do not provide clinically relevant results, and they cannot capture the stochastic character of neural dynamics.

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A strategy for designing valid standard addition calibrations and for optimising their uncertainty is presented. The design of calibrations involves the development of models of the sensitivity and precision of the instrumental signal, in a wide range of analyte concentration (or any other studied quantity), and the definition of sample dilution and standard addition procedures that allow fulfilling the assumptions of the linear unweighted regression model in, typically, a smaller range of standard addition calibrations. Calibrators are prepared by diluting the sample and adding analyte with negligible uncertainty to fit in a concentration range where signals are homoscedastic.

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The integration of up- and downstream unit operations can result in the elimination of hold steps, thus decreasing the footprint, and ultimately can create robust closed system operations. This type of design is desirable for the bioprocess of human mesenchymal stem cells (hMSC), where high numbers of pure cells, at low volumes, need to be delivered for therapy applications. This study reports a proof of concept of the integration of a continuous perfusion culture in bioreactors with a tangential flow filtration (TFF) system for the concentration and washing of hMSC.

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A simple, yet efficient, two-column simulated moving-bed (2CSMB) process for purifying adenovirus serotype 5 (Ad5) by size-exclusion chromatography (SEC) is presented and validated experimentally, and a general procedure for its robust design under parameter uncertainty is described. The pilot-scale run yielded a virus recovery of 86 percent and DNA and HCP clearances of 90 and 89 percent, respectively, without any fine tuning of the operating parameters. This performance compares very favorably against that of single-column batch chromatography for the same volume of size-exclusion resin.

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MicroRNAs (miRNAs) have emerged as a class of small, endogenous, regulatory RNAs that exhibit the ability to epigenetically modulate the translation of mRNAs into proteins. This feature enables them to control cell phenotypes and, consequently, modify cell function in a disease context. The role of inflammatory miRNAs in Alzheimer's disease (AD) and their ability to modulate glia responses are now beginning to be explored.

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Background: Performance indicators assessing the quality of medical care and linked to pay for performance may cause disagreement. Portuguese indicators included in recent health care reform are controversial.

Objectives: To obtain consensus from opinion leaders in family medicine regarding the performance indicators for practice management used in the evaluation of Family Health Units in Portugal.

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Adenovirus serotype 5 (Ad5) was successfully separated by size-exclusion chromatography (SEC) using a simple, yet efficient, two-column, quasi-continuous, simulated moving-bed process operated in an open-loop configuration. The operating cycle is divided into two identical half-cycles, each of them consisting of the following sequence of sub-steps: (i) elution of the upstream column and direction of the effluent of the downstream column to waste; (ii) elution of the upstream column and redirection of its effluent to waste while the downstream column is fed with the clarified bioreaction bulk and its effluent collected as purified product; (iii) operation of the system as in step (i) but collecting the effluent of the downstream column as product; (iv) elution of the upstream column and direction of its effluent to waste while the flow through the downstream column is temporarily halted. Clearance of impurities, namely DNA and host cell protein (HCP), were experimentally assessed.

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We present a new class of multicolumn chromatographic processes that change the classical way of handling the product outlets of simulated moving-bed (SMB) chromatography to avoid the use of flow controllers or an extra pump-the objective is to have just two- or three-way valves at a column outlet-while maintaining the analogy with the SMB in terms of displaced volumes of fluid per switch interval. In this class of processes the flow through a zone (or column) is always in one of the three states: (i) frozen, (ii) completely directed to the next zone (or column), or (iii) entirely diverted to a product line. We use the term relayed stream to refer to this particular type of manipulation of the outflow from a column.

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Chemical oxygen demand (COD) is one of the most relevant chemical parameters for the management of wastewater treatment facilities including the control of the quality of an effluent. The adequacy of decisions based on COD values relies on the quality of the measurements. Cost effective management of the minor sources of uncertainty can be applied to the analytical procedure without affecting measurement quality.

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Ecological theory of habitat heterogeneity and limited niche-similarity assumes that more heterogeneous environments provide a greater amount and diversity of resources than simple environments, resulting in a greater diversity of species. This study aimed to evaluate the effect of the habitat heterogeneity on the richness of dung beetles and to examine the spatial patterns of assemblage structure in relation to patterns of habitat heterogeneity. Dung beetles were collected using pitfall traps without bait in 30 points distributed in an area of cerrado sensu lato, in the region of Tangará da Serra, MT, Brazil, including areas of cerrado sensu stricto, campo sujo, cerradão and gallery forest.

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A comprehensive description of a new process--the GSSR (Gradient with Steady State Recycle) process--for center-cut separation by solvent-gradient chromatography is provided, highlighting its versatility, flexibility, and ease of operation. The GSSR process is particularly suited for ternary separation of bioproducts: it provides three main fractions or cuts, with a target product contained in the intermediate fraction. The process comprises a multicolumn, open-loop system, with cyclic steady state operation, that simulates a solvent gradient moving countercurrently with respect to the solid phase.

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Testing safety of foodstuffs of plant origin involves the analysis of hundreds of pesticide residues. This control is only cost-effective through the use of methods validated for the analysis of many thousands of analyte/matrix combinations. Several documents propose representative matrices of groups of matrices from which the validity of the analytical method can be extrapolated to the represented matrices after summarised experimental check of within group method performance homogeneity.

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We report on a numerical and experimental study of two-column versions of streamlined, multicolumn, semi-continuous chromatography for binary separation. The systems combine a flexible node design, cyclic flow-rate modulation, and relayed operation of the inlet/outlet ports to extend the mass-transfer zone over the largest possible length, while keeping it inside the system at all times. One advantage of these streamlined designs is the simplicity of their physical realization: regardless of the number of columns, they only require two pumps to supply feed and desorbent into the system, while the flow rates of liquid withdrawn from the system are controlled by material balance using simple two-way valves.

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A general methodology for a systematic evaluation of the uncertainty was derived for each particular ion in stock combined calibration standards in which concentrations of different ions extend over up to five orders of magnitude resulting in detailed uncertainty budgets with the aim of recognising the major contributions to combined uncertainties. This work confirmed that it is justifiable that the mass fraction of impurities in other chemicals is taken into account when calculating the mass concentration of an ion in combined calibration standard solution similarly to what is already common practice in accounting for the purity of chemical. It was proven that impurities in chemicals which are sources of major ions have significant effect on uncertainty budget of minor ions; already if the major ion exceeds 25 times the minor ion's concentration.

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Five polyphosphazenes with different hydrophilicites were synthesized and screened in vitro. The purpose was to identify unique types of polymeric substrates that distinctly favored or markedly prevented cellular adhesion. The SK-N-BE(2c) human neuroblastoma cell line, utilized for its electrogenic responses, was used to test this differential adhesion.

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