The development of sustainable protein sources is imperative for addressing the global challenge of food/feed security. Microalgae, which may be sustainably cultivated, are a promising source of proteins, gaining a progressive acceptance among consumers. The purpose of this work is to study the recovery of the protein C-phycocyanin from the microalga Arthrospira platensis (Spirulina), using a biphasic extraction system composed of sustainable solvents. The extraction system studied involves a feed phase, consisting of an aqueous salt solution and the target protein, and an extracting phase composed of a Natural Deep Eutectic Solvent (NADES) with affinity to the target protein. The performance of a specific NADES depends on the characteristics of the components of the NADES, in terms of its hydrophobicity/hydrophilicity balance, aiming the highest possible partitioning coefficient towards C-phycocyanin. It is also important to assure that the NADES phase selected presents a moderate viscosity and leads to a stable interface when in contact with the aqueous feed phase (i.e., presenting a measurable interfacial tension). In this work, after an extensive screening work of more than 71 combinations, the most overall performing combination is presented. This system shows a high partitioning coefficient of 29.4 ± 0.3 and an extraction yield of 99 % for C-phycocyanin (C-PC), demonstrating that the salting-out effect of the phosphate buffer and the hydrophobic character of NADES play a key role in this protein partitioning and recovery.

Alzheimer's disease (AD) is a devastating syndrome that accounts for 60-70 % of all dementia cases, putting an enormous burden on global healthcare and economy. Unfortunately, there is no cure for AD, and the currently approved drugs are limited in their effects. Given the various pathological mechanisms behind AD, the ``one-target, one-drug{''} paradigm for drug design became obsolete, and a new paradigm, polypharmacology, emerged. Consequently, a greater focus has been put towards multi-target directed ligands (MTDLs), as these can regulate several targets operating in the disease network. Parallel to that, cholinesterase inhibitors have regained popularity after decades of being considered only symptomatic agents with no disease-modifying properties. In this review, the current AD hypotheses and therapeutic targets, the concept of polypharmacology in AD pathology and the importance of cholinesterases in the pathogenesis and biochemical processes of AD are discussed, with a final overview of the current development in cholinesterase-based MTDLs.

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Dicarboxymethyl cellulose (DCMC) was synthesized and tested for protein adsorption. The prepared polymer was characterized by inductively coupled plasma atomic emission spectrometry (ICP-AES), attenuated total reflection Fourier-transform infrared spectroscopy (ATR-FTIR) and solid state nuclear magnetic resonance (ssNMR) to confirm the functionalization of cellulose. This work shows that protein adsorption onto DCMC is charge dependent. The polymer adsorbs positively charged proteins, cytochrome C and lysozyme, with adsorption capacities of 851 and 571 mg g(-1), respectively. In both experiments, the adsorption process follows the Langmuir adsorption isotherm. The adsorption kinetics by DCMC is well described by the pseudo second-order model, and adsorption equilibrium was reached within 90 min. Moreover, DCMC was successfully reused for five consecutive adsorption-desorption cycles, without compromising the removal efficiency (98-99%).

Stimuli-responsive supramolecular receptors are important building blocks for the construction of self-assembled functional materials. We report the design and synthesis of a pH and light-responsive 2-hydroxychalcone-beta-cyclodextrin conjugate (1-Ct) and its characterization by spectroscopic and computational methods. 1-Ct follows the typical reaction network of trans-chalcone-flavylium photoswitches. Upon light irradiation, 1-Ct can be photochemically converted into the cis-chalcone/hemiketal forms (1-Cc/1-B) under neutral pH conditions or to the flavylium cation (1-AH+) at acidic pH values. This stimuli-responsive beta-cyclodextrin host, 1Ct, was found to form stronger intramolecular self-inclusion complexes (Kintra = 14) than 1-AH+ (Kintra = 3) and weaker than 1-Cc/1-B (overall Kintra = 179), allowing control over their stability and binding properties by combinations of pH and light stimuli.

The eucalyptus weevil, Gonipterus spp. Schoenherr, 1833 (Coleoptera, Curculionidae) is considered a major pest of eucalyptus plantations. In regions where control is achieved, success is usually brought by the action of the solitary egg parasitoid Anaphes nitens (Girault, 1928) (Hymenoptera, Mymaridae). Research was conducted to ascertain which cues might mediate female wasp host location and selection. In Petri dish arenas, females were attracted to Gonipterus platensis Marelli, 1927 egg capsules, to G. platensis mated female faeces and to leaves of Eucalyptus globulus Labillardiere, 1799. Gas chromatography-mass spectrometry electroantennographic detection analysis was conducted using extracts obtained from leaves of E. globulus, from G. platensis egg capsules, as well as from adults of both sexes and their faeces, in order to detect and identify compounds perceived by the wasp's olfactory system. The parasitoids were shown to detect a wide range of compounds emitted by each one of these sources, and for 31 compounds, antennal response was confirmed by dose-response tests. Further behavioural trials were conducted in Petri dishes in order to decode the effect, on parasitoid behaviour, of selected compounds emitted by E. globulus and of the pheromones, emitted by the weevils on parasitoid behaviour. Attraction was observed to two compounds emitted by E. globulus, namely 1,8-cineole and gamma-terpinene, and to the main component of the male sex/aggregation pheromone, cis-verbenol. To our knowledge, this is the first report of attraction of a parasitoid from the family Mymaridae to a component of its host's sexual/aggregation pheromone. Similarly, to other egg parasitoid species, A. nitens females are likely to use the host plant volatiles as long-range host location cues and to adopt the `infochemical detour' strategy in order to get in the vicinity of their hosts.

The eucalyptus weevil, Gonipterus platensis (Coleoptera, Curculionidae), is a major pest of eucalyptus plantations worldwide. To date, no pheromones have been identified for this species, despite their valuable potential as tools in monitoring or control strategies. Here we report the detection and identification of pheromones candidates of G. platensis. The weevil's volatile compounds were collected by solid phase micro extraction (SPME) and monolithic material sorption extraction (MMSE). Using Gas Chromatography coupled to Mass Spectrometry (GC/MS) analysis, eleven insect specific compounds were detected and identified: verbenene, cis-verbenol, trans-verbenol, verbenone, 2-oxo-1,8-cineole, 9-hydroxy-1,8-cineole, 2-alpha-hydroxy-1,8-cineole, 3-oxo-1,8-cineole, 2-beta-hydroxy-1,8-cineole, 3-alpha-hydroxy-1,8-cineole and 7-hydroxy-1,8-cineole. Three of these compounds, verbenene, cis-verbenol and trans-verbenol, were shown to be male-specific. Antennal sensitivity towards ten compounds emitted by G. platensis was detected using Gas Chromatography-Mass Spectrometry/Electroantennographic Detection (GC-MS/EAD). Extracts from virgin males proved to be attractive to virgin females in olfactometer bioassays. Further behavioural bioassays showed that both virgin females and virgin males were attracted to the male-specific compound cis-verbenol and that virgin females were attracted to trans-verbenol. Verbenone was attractive to mated females. Regarding 2-alpha-hydroxy-1.8-cineole and 2-oxo-1,8-cineole, which are produced by both sexes, the alcohol was attractive to virgin males and both the alcohol and the ketone were repellant to mated females. This is, to our knowledge, the first identification of pheromones candidates in Gonipterus spp. and also the first evidence of cineole metabolites acting as semiochemicals.

A missense mutant of a Dps protein (DNA-binding protein from starved cells) from Marinobacter hydrocarbonoclasticus was used as a building block to develop a new supramolecular assembly complex which enhances the iron uptake, a physiological function of this mini-ferritin. The missense mutation was conducted in an exposed and flexible region of the N-terminal, wherein a threonine residue in position 10 was replaced by a cysteine residue (DpsT10C). This step enabled a click chemistry approach to the variant DpsT10C, where a thiol-ene coupling occurs. Two methods and two types of linker were used resulting in two different mini-ferritin supramolecular polymers, which have maintained secondary structure and native iron uptake physiological function. Electrophoretic assays and mass spectrometry were utilized to confirm that both functionalization and coupling reactions occured as predicted. The secondary structure has been investigated by circular dichroism and synchrotron radiation circular dichroism. Size and morphology were obtained by dynamic light scattering, size exclusion chromatography and atomic force microscopy, respectively. The iron uptake of the synthesized protein polymers was confirmed by UV-Vis spectroscopy loading assays.