One‐Step Formulation of Protein Microparticles with Tailored Properties: Hard Templating at Soft Conditions

Formulation of therapeutic proteins into particulate forms is a main strategy for site‐specific and prolonged protein delivery as well as for protection against degradation. Precise control over protein particle size, dispersity, purity, as well as mild preparation conditions and minimal processing steps are highly desirable. It is, however, hard to fit all these criteria with conventional preparation techniques. Here a one‐step hard‐templating synthesis of microparticles composed of functional, non‐denatured protein is reported. The method is based on filling porous CaCO₃ microtemplates with the protein near to its isoelectric point (pI) followed by pH‐ or EDTA‐mediated dissolution of the tempplates. In principle, a wide variety of proteins can be converted into microparticles using this approach. The main requirement is an overlap of the protein insolubility and a template solubility for a certain parameter (here pH or EDTA). Here the formulation of insulin particles is studied in detail and it is shown that particles consisting of high molecular weight protein (catalase) can also be prepared. In this context, the synthesis of CaCO₃ templates with controlled size, the mechanism of the protein microparticle formation and mechanical properties of the microparticles are discussed. For the first time, the fabrication of mesoporous monodispersed CaCO₃ microtemplates with identical porocity but tuned diameter from 3 to 20 μm is demonstrated. The protein particle diameter can be adjusted by choosing the appropriate template size that is critical for successful pulmonary delivery of insulin. As a first step towards insulin delivery, the in vitro release of insulin at physiological conditions is studied.     

Analytical Tools Used to Distinguish Chemical Profiles of Plants Widely Consumed as Infusions and Dietary Supplements: Artichoke,Milk Thistle,and Borututu

Artichoke, borututu, and milk thistle are three medicinal plants widely consumed as infusions or included in dietary supplements (e.g., pills and syrups). Despite the high consumption of these plants, studies on their nutritional value and primary metabolites are scarce, with their composition in secondary metabolites such as phenolic compounds being only reported. Therefore, their nutritional value was assessed, and analytical tools (liquid and gas chromatography coupled to different detectors) were used to distinguish their chemical profiles according to their hydrophilic (sugars and organic acids) and lipophilic (fatty acids and tocopherols) components. Chromatographic techniques are important analytical tools used in the identification and quantification of several molecules, also being a standard requirement to distinguish different profiles. Borututu gave the highest energetic value with the highest content of carbohydrates and fat, sucrose and total sugars, shikimic and citric acids, and α-, β-, δ-, and total tocopherols. Artichoke had the highest ash and protein contents, oxalic acid, SFA (mainly palmitic acid), and γ-tocopherol, and also the best n-6/n-3 ratio. Milk thistle showed the highest levels of fructose and glucose, quinic acid and total organic acids, and PUFA, mainly linoleic acid, and the best PUFA/SFA ratio. The hydrophilic compounds identified in the studied plants, mostly sugars, are responsible for the energy contribution of their widely consumed infusions. Otherwise, the bioactivity of lipophilic compounds, namely, unsaturated fatty acids and tocopherols, is lost in those preparations but can be recovered in dietary supplements based on the plants. As far as we know, this is the first report on the detailed composition of the three plants regarding their nutritional features.     

Gain scheduling adaptive control applied to a particular mixer-settler equipment

The mixer-settler based on phase inversion (MSBPI) is a system used for treatment of wastewaters in order to reduce the content of oils and greases from wastewaters produced in oil industry. Its operation principle combines mixing, extraction and separation by decantation in a unique process. In particular, for control purpose, the level of the interface between organic solvent and water, in the separation section, is a critical variable to guarantee performance and security during operation of the MSBPI. In this work, an image-based detector to monitor the organic solvent–water interface level is proposed and a feedback control loop for the process is presented. Initially, a conventional controller (PID) was implemented and, subsequently, a strategy of gain scheduling adaptive control was developed in order to improve the process response in closed-loop. The experimental results showed that the adaptive control presented an efficient performance, even when disturbances of significant magnitude were applied to the process.     

Application of full-factorial design for optimization of the moisture determination in wheat flour using infrared scales

We studied the parameters that may affect the analysis of the moisture content in wheat flour using infrared scales. The parameters were studied by one-factor at a time methodology and then, by full-factorial design (2³). The best condition using infrared scales occurred at 120 °C, with a sample weight of 1.5 g and an exposure time to infrared radiation of 6 min. We observed the interaction effect between the three factors studied. The analysis time was 60 times lower than classical method. The optimized method for determination of the moisture content in wheat flour using infrared scales was validated to range between 9 and 17 %.     

In vitro intestinal absorption of amino acid mixtures extracted from codfish (Gadus morhua L.) salting wastewater

This study reports on the intestinal permeability of salt‐containing mixtures of amino acids extracted from codfish salting wastewater. Permeability was evaluated in vitro using the Caco‐2 cell line model; cell integrity during exposure to mixtures of amino acids was estimated by measuring the transepithelial electrical resistance (TEER). The effect of salt (NaCl) on the permeability and on the intestinal cell's integrity was also examined. Permeation rate (i.e. transport) was ≥95% for all amino acids except for creatine, for which it was 6%. Values for apparent permeability coefficients, P_app > 10^?5 cm s^?1, for mixture with isotonic concentration of NaCl suggest that amino acids are very likely to be absorbed in humans. Mixture with a hypertonic level of NaCl exerts a cytotoxic effect in intestinal cells resulting in a loss of epithelium integrity. Results show that isotonic mixture of amino acids extracted from codfish salting wastewater could be used in food, feed, cosmetic and pharmaceutical formulations. These applications could contribute to the fish industry sustainability.     

Automatic word naming recognition for an on-line aphasia treatment system

One of the most common effects among aphasia patients is the difficulty to recall names or words. Typically, word retrieval problems can be treated through word naming therapeutic exercises. In fact, the frequency and the intensity of speech therapy are key factors in the recovery of lost communication functionalities. In this sense, speech and language technology can have a relevant contribution in the development of automatic therapy methods. In this work, we present an on-line system designed to behave as a virtual therapist incorporating automatic speech recognition technology that permits aphasia patients to perform word naming training exercises. We focus on the study of the automatic word naming detector module and on its utility for both global evaluation and treatment. For that purpose, a database consisting of word naming therapy sessions of aphasic Portuguese native speakers has been collected. In spite of the different patient characteristics and speech quality conditions of the collected data, encouraging results have been obtained thanks to a calibration method that makes use of the patients’ word naming ability to automatically adapt to the patients’ speech particularities.     

Microencapsulation of Propolis in Protein Matrix Using Spray Drying for Application in Food Systems

Propolis presents several health benefits due to the presence of bioactive compounds, mainly phenolic compounds; however, its application in food is limited due to undesirable odor and low water solubility. The bioactive compounds are usually susceptible to degradation by exposure to light, heat, or oxygen or by interaction with other compounds, which may limit its biological activity. The study aimed the propolis extract microencapsulation using rice, pea, soybean, and ovoalbumin proteins as wall material by spray drying and to analyze their in vitro digestion. The propolis extract presented a high concentration of apigenin. Encapsulation efficiency was greater than 70%, and it was maintained the antioxidant activity of propolis (88% inhibition of DPPH for propolis extract and >?73% for the microparticles). The DSC, ATR-FTIR, and X-ray diffraction techniques confirmed the encapsulation. The microparticles showed different shapes, sizes, and physical characteristics. The microparticles encapsulated with pea protein could be used in formulations of Minas Frescal cheese due to the controlled released, whereas the other microparticles could be used in pudding formulations.