Development of water-in-oil emulsions as delivery vehicles and testing with a natural antimicrobial extract

Water-in-oil (W/O) emulsions have high potential for several industrial areas as delivery systems of hydrophilic compounds. In general, they are less studied than oil-in-water (O/W) systems, namely in what concerns the so-called fluid systems, partly due to problems of instability. In this context, this work aimed to produce stable W/O emulsions from a natural oil, sweet almond oil, to be further tested as vehicles of natural hydrophilic extracts, here exemplified with an aqueous cinnamon extract. Firstly, a baseW/O emulsion using a high-water content (40/60, v/v) was developed by testing di erent mixtures of emulsifiers, namely Tween 80 combined with Span 80 or Span 85 at di erent contents. Among the tested systems, the one using a 54/46 (v/v) Span 80/Tween 80 mixture, and subjected to 12 high-pressure homogenizer (HPH) cycles, revealed to be stable up to 6 months, being chosen for the subsequent functionalization tests with cinnamon extract (1.25–5%; w/v; water-basis). The presence of cinnamon extract leaded to changes in the microstructure as well as in the stability. The antimicrobial and antioxidant analysis were evidenced, and a sustained behavior compatible with an extract distribution within the two phases, oil and water, in particular for the higher extract concentration, was observed.Base Funding—UIDB/00690/2020 of CIMO—Mountain Research Center—funded by national funds through FCT/MCTES (PIDDAC). Base Funding—UIDB/50020/2020 of the Associate Laboratory LSRE-LCM—funded by national funds through FCT/MCTES (PIDDAC).info:eu-repo/semantics/publishedVersio

Spirulina (Arthrospira platensis) protein-rich extract as a natural emulsifier for oil-in-water emulsions: optimization through a sequential experimental design strategy

Spirulina (Arthrospira platensis) proteins have been proven to present emulsifying properties. In this work, a Spirulina protein-rich extract obtained by ultrasound extraction (SpE) was tested to stabilize oil-in-water (O/W) emulsions. For this purpose, a sequential experimental design strategy (Fractional Factorial Design (FFD) 24–1 followed by a Central Composite Rotatable Design (CCRD) 22)) was applied. The effect of four variables, SpE concentration, O/W weight ratio, pH and storage time, on emulsions’ zeta potential and number-mean droplet diameter was considered for the FFD 24–1, indicating SpE concentration and storage time as the relevant variables for the CCRD 22. According to zeta potential and number-mean droplet diameter evaluation, for the studied SpE concentration range (2–5 wt%), quite stable emulsions were obtained along the tested 30-days period. Even so, for 5%, visual inspection revealed extract segregation after 20-days. The optimal solution comprised 4 wt% of SpE, for an O/W weight ratio of 30/70 and a pH of 7.0 (number-mean droplet diameter of 55.66 nm and zeta potential of -43.83 mV). Overall, SpE has proven to be an excellent emulsifier, offering the potential to substitute animal-based proteins and synthetic emulsifiers. In addition, no signs of contamination by microorganisms were observed, suggesting that the SpE may also act as an antimicrobial agent.CIMO (Centro de Investigação de Montanha, Portugal) (UIDB/00690/2020), LSRE-LCM (Laboratory of Separation and Reaction Engineering – Laboratory of Catalysis and Materials, Portugal) (UIDB/50020/2020; UIDP/50020/2020), and ALiCE (Associate Laboratory in Chemical Engineering, Portugal) (LA/P/0045/2020), funded by national funds through Fundação para Ciência e a Tecnologia/Ministério da Ciência, Tecnologia e Ensino Superior (FCT/MCTES) (PIDDAC). National funding by Fundação para a Ciência e a Tecnologia (FCT) (Portugal), P.I., through the institutional scientific employment program contract with A. Santamaria-Echart, L. Barros, and A. Fernandes. FCT for the PhD research grant of Samara Cristina da Silva (SFRH/BD/148281/2019) and Giovana Colucci (2021. 05215. BD).info:eu-repo/semantics/publishedVersio

New Trends in Natural Emulsifiers and Emulsion Technology for the Food Industry

The food industry depends on using different additives, which increases the search for effective natural or natural-derived solutions, to the detriment of the synthetic counterparts, a priority in a biobased and circular economy scenario. In this context, different natural emulsifiers are being studied to create a new generation of emulsion-based products. Among them, phospholipids, saponins, proteins, polysaccharides, biosurfactants (e.g., compounds derived from microbial fermentation), and organic-based solid particles (Pickering stabilizers) are being used or start to gather interest from the food industry. This chapter includes the basic theoretical fundamentals of emulsions technology, stabilization mechanisms, and stability. The preparation of oil-in-water (O/W) and water-in-oil (W/O) emulsions, the potential of double emulsions, and the re-emerging Pickering emulsions are discussed. Moreover, the most relevant natural-derived emulsifier families (e.g., origin, stabilization mechanism, and applications) focusing food applications are presented. The document is grounded in a bibliographic review mainly centered on the last 10-years, and bibliometric data was rationalized and used to better establish the hot topics in the proposed thematic

Evaluation of micro-energy dispersive X-ray fluorescence and histochemical tests for aluminium detection in plants from High Altitude Rocky Complexes, Southeast Brazil

The soils developed under High Altitude Rocky Complexes in Brazil are generally of very low chemical fertility, with low base saturation and high exchangeable aluminium concentration. This stressful condition imposes evolutionary pressures that lead to ecological success of plant species that are able to tolerate or accumulate high amounts of aluminium. Several analytical methods are currently available for elemental mapping of biological structures, such as micro-X-ray fluorescence (μ-EDX) and histochemical tests. The aim of this study was to combine μ-EDX analysis and histochemical tests to quantify aluminium in plants from High Altitude Rocky Complexes, identifying the main sites for Al-accumulation. Among the studied species, five showed total Al concentration higher than 1000 mg kg−1. The main Al-hyperaccumulator plants, Lavoisiera pectinata, Lycopodium clavatum and Trembleya parviflora presented positive reactions in the histochemical tests using Chrome Azurol and Aluminon. Strong positive correlations were observed between the total Al concentrations and data obtained by μ-EDX analysis. The μ-EDX analysis is a potential tool to map and quantify Al in hyperaccumulator species, and a valuable technique due to its non-destructive capacity. Histochemical tests can be helpful to indicate the accumulation pattern of samples before they are submitted for further μ-EDX scrutiny

The powerful neuroprotective action of C1-inhibitor on brain ischemia-reperfusion injury does not require C1q

C1-inhibitor (C1-INH) is a major regulator of the complement classical pathway. Besides this action, it may also inhibit other related inflammatory systems. We have studied the effect of C1-INH in C57BL/6 mice with focal transient brain ischemia induced by 30 minutes of occlusion of the middle cerebral artery. C1-INH induced a dose-dependent reduction of ischemic volume that, with the dose of 15 U/mouse, reached 10.8% of the volume of saline-treated mice. Four days after ischemia the treated mice had significantly lower general and focal neurological deficit scores. Fluoro-Jade staining, a marker for neuronal degeneration, showed that C1-INH-treated mice had a lower number of degenerating cells. Leukocyte infiltration, as assessed by CD45 immunostaining, was also markedly decreased. We then investigated the response to ischemia in C1q-/- mice. There was a slight, nonsignificant decrease in infarct volume in C1q-/- mice (reduction to 72.3%) compared to wild types. Administration of C1-INH to these mice was still able to reduce the ischemic volume to 31.4%. The study shows that C1-INH has a strong neuroprotective effect on brain ischemia/reperfusion injury and that its action is independent from C1q-mediated activation of classical pathway

Pre-clinical evidence for an anti-tumor activity of defibrotide, a DNA-based, endothelium stabilizing drug

14144 Background: Defibrotide (DF) is a mixture of polydeoxyribonucleotides with anti-thrombotic activity. Next to endothelium stabilization, recent data suggest anti-neoplastic properties of DF modulating interactions of tumor cells with their microenvironment. We investigated whether DF regulates expression and activity of heparanase, an enzyme critically involved in breaking down extracellular barriers and releasing growth factors linked to tumor invasion and angiogenesis. Methods: Heparanase expression was tested by RT-PCR and flow cytometry with multiple myeloma (MM) and microvascular endothelial cells. Heparanase activity was measured in cellular extracts with a heparan-degrading enzymatic assay. Serum degradation products of DF were identified by SEC-HPLC. The anti-angiogenic potential of DF was tested in vitro using a kit with human microvascular endothelial cells forming tubes across a layer of fibroblasts. In vivo, DF was tested in the dorsal skin-fold chamber assay in mice after inoculation of human gastric cancer cells. Proliferation was assessed by trypan blue exclusion. Results: We demonstrate a striking downregulation of expression and enzymatic activity of heparanase in endothelial as well as MM cells. In contrast, the degradation products of DF failed to exert any biological activity, suggesting that the intact mixture of deoxyoligonucleotides is responsible for the anti-tumor effect. We could also show that DF prevents (tumor) angiogenesis in vitro and in vivo. Western blots suggest that DF reduces phosphorylation-activation of p70S6 kinase, a key target in the mTOR pathway linked to angiogenesis. In addition, DF does not influence proliferation of vascular or tumor cells, rather acts via selective inhibition of tube formation of endothelial cells. Conclusion: In the present report we provide evidence for an anti-tumor activity of DF. DF inhibits (tumor) vessel formation and heparanase activity, and thus should be considered as an anti-cancer agent. [Table: see text] </jats:p