Microstructure, Physical Properties, and Oxidative Stability of Olive Oil Oleogels Composed of Sunflower Wax and Monoglycerides.

The utilization of natural waxes to form oleogels has emerged as a new and efficient technique for structuring liquid edible oil into solid-like structures for diverse food applications. The objective of this study was to investigate the interaction between sunflower wax (SW) and monoglycerides (MGs) in olive oil oleogels and assess their physical characteristics and storage stability. To achieve this, pure SW and a combination of SW with MGs in a 1:1 ratio were examined within a total concentration range of 6-12% w/w. The formed oleogels were characterized based on their microstructure, melting and crystallization properties, textural characteristics, and oxidative stability during storage. All the oleogels were self-standing, and, as the concentration increased, the hardness of the oleogels also increased. The crystals of SW oleogels were long needle-like, while the combination of SW and MGs led to the formation of crystal aggregates and rosette-like crystals. Differential scanning calorimetry and FTIR showed that the addition of MGs led to different crystal structures. The oxidation results revealed that oleogels had low peroxide and TBARS values throughout the 28-day storage period. These results provide useful insights about the utilization of SW and MGs oleogels for potential applications in the food industry.

A floating 3D printed polypill formulation for the coadministration and sustained release of antihypertensive drugs.

Polypharmacy is a common issue, especially among elderly patients resulting in administration errors and patient inconvenience. Hypertension is a prevalent health condition that frequently leads to polypharmacy, as its treatment typically requires the co-administration of more than one different Active Pharmaceutical Ingredients (API's). To address these issues, floating hollow torus-shaped dosage forms were developed, aiming at providing prolonged gastric retention and sustained drug release. The dosage forms (polypills) containing three anti-hypertensive API's (diltiazem (DIL), propranolol (PRP) and hydrochlorothiazide (HCTZ)) were created via Fused Deposition Modelling 3D printing. A multitude of the dosage forms were loaded into a capsule and the resulting formulation achieved prolonged retention times over a 12-hour period in vitro, by leveraging both the buoyancy of the dosage forms, and the "cheerios effect" that facilitates the aggregation and retention of the dosage forms via a combination of surface tension and shape of the objects. Physicochemical characterization methods and imaging techniques were employed to investigate the properties and the internal and external structure of the dosage forms. Furthermore, an ex vivo porcine stomach model revealed substantial aggregation, adhesion and retention of the 3D printed dosage forms in porcine stomach. In vitro dissolution testing demonstrated almost complete first-order release of PRP and DIL (93.52 % and 99.9 %, respectively) and partial release of HCTZ (65.22 %) in the 12 h timeframe. Finally, a convolution-based single-stage approach was employed in order to predict the pharmacokinetic (PK) parameters of the API's of the formulation and the resemblance of their PK behavior with previously reported data.

Cereal-Based 3D Printed Dosage Forms for Drug Administration During Breakfast in Pediatric Patients within a Hospital Setting.

In an effort to combine a child-friendly dosage form for medication administration in hospitalized pediatric patients and a user-friendly automated process for its preparation by health-care providers, the current study proposes a method for drug administration with breakfast using semi-solid extrusion 3D printing. Cereal was used as the platform carrier of the hydrophobic ibuprofen and the hydrophilic paracetamol to develop the drug loaded cereal ink. Rheological analysis was performed to identify the cereal ink with optimum viscosity for extrusion printing. Drug distribution and crystallinity within the printed cereal were assessed with confocal Raman microscopy and thermal and X-ray diffraction analysis, respectively, indicating molecular dispersion of both drugs within the cereal. High cereal porosity was associated with a higher milk absorption capacity and a decrease in their flexural force upon immersion in milk. Dissolution studies were performed in biorelevant media under fasted and fed state conditions and in the presence of full-fat and low-fat milk showing dissolution enhancement of the poorly soluble ibuprofen in the presence of the higher fat content milk. Concealing drug administration under the auspice of this essential daily eating habit is expected to facilitate overcoming adherence barriers to medication intake by pediatric patients within a hospital setting.

Stability of natural food colorants derived from onion leaf wastes.

Plant anthocyanins have widely been employed as natural food colorants. However, their instability restricts many of their applications in food industry. In this study, anthocyanins were extracted from onion outer scales, using aqueous solutions with or without added cyclodextrins (CDs). The results indicated that when cyclodextrins were included in the extraction medium, the anthocyanins were stable or even had improved and augmented color intensity upon storage or following thermal and UV-light treatments over a broad pH range (2.0-7.0). FT-IR and UV-Vis spectroscopy measurements confirmed the formation of inclusion complexes between CDs and anthocyanins and the presence of pyranosyl groups (pyranoanthocyanins) upon heating. Overall, the stability of onion anthocyanins under various environmental stresses, often encountered during food processing and storage, indicates that the natural color extract from onion outer scales can be of value as an interesting colorant alternative for food applications.

Physicochemical properties of human breast milk during the second year of lactation.

The present study examined the microstructure as well as the physicochemical properties of human milk during the second year of lactation in an attempt to explore its applicability for the formulation of food products. It was observed that human milk fat globules (MFG) droplet size increased within 3 days of milk extraction due to coalescence, as evidenced by confocal microscopy. Furthermore, a gradual decrease of the average MFG size was noted from the sixteenth (16th) to twenty-fifth (25th) month of lactation. It was also found that the size of casein micellar structures increased upon acidification to pH 4.3 (isoelectric point of human caseins). In addition, human milk proteins enhanced the stability of oil-in-water emulsions against coalescence compared to cow, sheep, and goat milk proteins employed as macromolecular emulsifying ingredients. The cold-acid-gels of human milk proteins showed a less elastic behavior than the other milk samples, possibly due to the different structure, composition and size of human casein micelles. Furthermore, the DSC thermograms showed that human whey proteins are denatured in the same temperature range as do the cow whey proteins, but exhibit different thermal transition profiles. Overall, the findings of this research confirm that both the structure and the physicochemical properties of human milk are affected by the stage of lactation. Moreover, the particular composition and structure of human milk proteins seem to be responsible for the special functional characteristics of human milk that may lead towards the formulation of innovative products.

Effects of Cornus and Its Mixture with Oregano and Thyme Essential Oils on Dairy Sheep Performance and Milk, Yoghurt and Cheese Quality under Heat Stress.

The effect of a diet supplemented with a novel cornus extract, enriched with essential oils of oregano and thyme, on the performance of Chios cross-bred dairy sheep was investigated during the summer period. The plant extracts were prepared using a "green" method based on aqueous extraction. A total of 45 lactating ewes were allocated into three equal groups in a randomized block design. The three groups were fed the same feed allowance, roughage based on Lucerne hay and wheat straw and a concentrate based on cereals and oil cakes (the control diet). The diet of two groups was fortified with cornus extract, with or without oregano and thyme essential oils, at a level 0.515 g of plant extract/essential oils per kg of concentrate. Individual milk yield was recorded weekly and feed refusals were recorded on a pen basis daily, during a six-week period of lactation. Milk samples were analyzed for the chemical composition of protein, fat, lactose and solids-not-fat constituents, somatic cell counts and total viable bacteria counts. Moreover, the milk of each group was used for yoghurt and Feta cheese production. The lipid oxidative stability, protein carbonyl content and fatty acid composition of milk, yoghurt and cheese samples were also evaluated. The results showed that the incorporation of novel plant extracts and essential oils increased the milk production per ewe. Dietary supplementation with cornus extracts and essential oils lowered lipid and protein oxidation in milk, yoghurt and cheese samples, compared to the control. However, diet supplementation with herbal extracts did not affect the fatty acid profile in milk, cheese and yoghurt or the serum biochemical parameters. In conclusion, dietary supplementation with cornus in combination with oregano and thyme has the potential to improve feed utilization and the performance of high-yield dairy Chios cross-bred ewes reared under heat stress.

Ocular co-Delivery of Timolol and Brimonidine from a Self-Assembling Peptide Hydrogel for the Treatment of Glaucoma: In Vitro and Ex Vivo Evaluation.

Effective pharmacotherapy during glaucoma treatment depends on interventions that reduce intraocular pressure (IOP) and retain the IOP lowering effect for sufficient time so as to reduce dosing frequency and enhance patient adherence. Combination anti-glaucoma therapy and dosage forms that increase precorneal residence time could therefore constitute a promising therapeutic intervention. The in-situ gel forming self-assembling peptide ac-(RADA)4-CONH2 was evaluated as carrier for the ocular co-delivery of timolol maleate (TM) and brimonidine tartrate (BR). The hydrogel's microstructure and mechanical properties were assessed with atomic force microscopy and rheology, respectively. Drug diffusion from the hydrogel was evaluated in vitro in simulated tear fluid and ex vivo across porcine corneas and its effect on the treated corneas was assessed through physicochemical characterization and histological analysis. Results indicated that TM and BR co-delivery affected hydrogel's microstructure resulting in shorter nanofibers and a less rigid hydrogel matrix. Rapid and complete release of both drugs was achieved within 8 h, while a 2.8-fold and 5.4-fold higher corneal permeability was achieved for TM and BR, respectively. No significant alterations were induced in the structural integrity of the corneas treated with the hydrogel formulation, suggesting that self-assembling peptide hydrogels might serve as promising systems for combination anti-glaucoma therapy.

Pediatric-friendly chocolate-based dosage forms for the oral administration of both hydrophilic and lipophilic drugs fabricated with extrusion-based 3D printing.

The aim of the current study was the development of pediatric-friendly 3D printed chocolate-based oral dosage forms. Corn syrup was used to both facilitate the incorporation of a lipophilic, namely ibuprofen or a hydrophilic, namely paracetamol, active compound that were used as model drugs and to enable 3D printing of the chocolate-based dosage forms. Physicochemical (differential scanning calorimetry, X-Ray diffraction, Fourier-Transform infrared spectroscopy, particle size distribution) and rheological studies were applied for the characterization of the prepared chocolate-based formulations. Texture profile analysis and in vitro digestion studies were performed in order to further analyze the texture attributes and to evaluate drug dissolution of the final dosage forms, respectively. In the present study, we reported on a facile method for the preparation of a 3D printed chewable chocolate-based dosage form with rapid and high release of both hydrophobic and hydrophilic drugs in simulated salivary fluid. The application of 3D printing technology enables accuracy in dose adjustment, while at the same time introducing the potential of patient's active involvement in customization of the design, textural and organoleptic properties of the final dosage form.

Whey proteins: Musings on denaturation, aggregate formation and gelation.

Whey proteins are globular milk proteins with numerous functional properties and a broad potential for usage in food and supplement-pharmaceutical products. Under various denaturing conditions, whey proteins unfold, form aggregates, microparticles, while at sufficiently high concentrations, gel networks are created. The continuously growing research interest makes it necessary to discuss the recent advances and achievements in this area, including the basic principles of the structural assembly's kinetics along with the different approaches undertaken to fabricate whey protein gels as well as the means to modulate the gel physical properties. This article also focuses on the use of ethanol to modify the structure and functionality of whey proteins, as an alternative way to change the protein conformation and thereby gelation phenomena and functionality.

Whole Genome Sequencing and Root Colonization Studies Reveal Novel Insights in the Biocontrol Potential and Growth Promotion by Bacillus subtilis MBI 600 on Cucumber.

Bacillus spp. MBI 600 is a gram-positive bacterium and is characterized as a PGPR strain involved in plant growth promotion and control of various plant pathogens which has recently been introduced into the agricultural practice. In this study we performed a Next Generation Sequencing analysis, to analyze the full genome of this microorganism and to characterize it taxonomically. Results showed that MBI 600 strain was phylogenetically close to other Bacillus spp. strains used as biocontrol agents and identified as B. subtilis. GOG analysis showed clusters contributed to secondary metabolites production such as fengycin and surfactin. In addition, various genes which annotated according to other plant-associated strains, showed that play a main role in nutrient availability from soil. The root colonization ability of MBI 600 strain was analyzed in vivo with a yellow fluorescence protein (yfp) tag. Confocal laser scanning microscopy of cucumber roots treated with yfp-tagged MBI 600 cells, revealed that the strain exhibits a strong colonization ability of cucumber roots, although it is affected significantly by the growth substrate of the roots. In vitro and in planta experiments with MBI 600 strain and F. oxysporum f.sp. radicis cucumerinum and P. aphanidernatum, showed a high control ability against these soilborne pathogens. Overall, our study demonstrates the effectiveness of MBI 600 in plant growth promotion and antagonism against different pathogens, highlighting the use of this microorganism as a biocontrol agent.

Natural food colourants derived from onion wastes: application in a yoghurt product.

The valorization of onion (Allium cepa) solid wastes, a 450,000 tonnes/year waste in Europe, by a green extraction method is presented. Polyphenols of onion solid wastes were extracted using eco-friendly solvents, such as water and glycerol. The 2-hydroxypropyl-ß-cyclodextrin was also used as a co-solvent for the augmentation of the extraction yield. The process has been optimized by implementing a central composite face centered design of experiments, with two replicates in the central point, taking into consideration the following independent variables: glycerol concentration, cyclodextrin concentration and temperature. The assessment of the extraction model was based on two responses: the total pigment yield and the antiradical capacity. LC-MS analysis was also employed in order to identify polyphenols and colourants of the obtained extracts. The main polyphenols found were quercetin and quercetin derivatives and the main colourant was cyanidin 3-O-glucoside. The extract was also tested as a food colourant in a yoghurt matrix. The onion leaf extract was found to be a stable natural colourant and could be utilized as an alternative ingredient to synthetic coloring agents. This article is protected by copyright. All rights reserved.

Microrheology and microstructure of water-in-water emulsions containing sodium caseinate and locust bean gum.

The mechanical response on the microscale of phase-separated water-in-water emulsions containing sodium caseinate (SCN) and locust bean gum (LBG) has been monitored by confocal laser scanning microscopy and particle tracking microrheology. Mixed biopolymer systems exhibiting phase-separated micro-regions were enriched in either protein or polysaccharide in the continuous or dispersed phase, depending on the weight ratio of the two biopolymers. Measurements of the tracking of charged probe particles revealed that the local rheological properties of protein-rich regions were considerably lower than that of LBG-rich domains for all the biopolymer ratios examined. At pH 7 in the absence of added salt, the viscosity of the protein-rich regions was little affected by an increase in overall LBG concentration, which is consistent with the phase separation mechanism in the mixed solution of charged (SCN) and uncharged (LBG) biopolymers being dominated by the relative entropy of the counter-ions associated with the charged protein molecules. Addition of salt was found to produce an enhancement in the level of thermodynamic incompatibility, leading to faster and more pronounced phase separation, and altering the micro-viscosity of protein-rich regions. At high ionic strength, it was also noted that there was a pronounced accumulation of incorporated probe particles at the liquid-liquid interface. The microrheological properties of the SCN-rich regions were found to be substantially pH-dependent in the range 7 > pH > 5.4. By adjusting the acidification conditions and the biopolymer ratio, discrete protein-based microspheres were generated with potential applications as a functional food ingredient.

Factors affecting migration kinetics from a generic epoxy-phenolic food can coating system.

This study investigated how the properties of a polymeric can coating film, such as thickness and crosslink density as well as the type of migrant, influence the migration kinetics of model migrants in an attempt to better understand, model and control the migration process. Four model migrants were used BADGE (bisphenol A diglycidyl ether), BADGE·H2O, cyclo-diBADGE and Uvitex OB, that differ in size and polarity. Fatty and aqueous food simulants were used at high temperatures (70-130°C). The apparent diffusion coefficients were found to decrease with increasing crosslink density, while they increased with increasing film thickness. The apparent activation energy of BADGE and BADGE-related compounds was calculated from the diffusion data and were high, in the range of 250-264kJmol-1. The polarity of the simulant and the polarity of the migrant were found to influence migration. The results can be used to improve existing migration models, and thereby help to reduce migration from packaging into food by using safety-by-design approaches in new product development.

Effect of heat, pH, ultrasonication and ethanol on the denaturation of whey protein isolate using a newly developed approach in the analysis of difference-UV spectra.

A newly developed method of analysis of difference-UV spectra was successfully implemented in the study of the effect of heat, pH, ultrasonication and ethanol on the denaturation of whey protein isolate. It was found that whey proteins exhibit their highest stability against heat denaturation at pH 3.75. At very low pH values, i.e. 2.5, they exhibited considerable cold denaturation, while after heating at this pH value, the supplementary heat denaturation rate was lower compared to that at neutral pH. The highest heat denaturation rates were observed at pH values higher than neutral. High power sonication on whey proteins, previously heated at 90°C for 30min, resulted in a rather small reduction of the fraction of the heat denatured protein aggregates. Finally, when ethanol was used as a cosolvent in the concentration range 20-50%, a sharp increase in the degree of denaturation, compared to the native protein solution, was observed.

Studying the denaturation of bovine serum albumin by a novel approach of difference-UV analysis.

A novel approach in the analysis of difference-UV spectrophotometric data for determining the heat denaturation degree of bovine serum albumin (BSA) was assessed. Five different parameters of difference-UV spectra were obtained by subtracting spectra of unheated and denatured protein solutions at different temperature-time combinations. BSA was found to exhibit a maximum degree of heat denaturation of about 17% compared to the complete unfolding caused by 6M guanidine hydrochloride. This low degree of heat denaturation is probably caused by the aggregation of the initially unfolded protein molecules. The kinetic analysis exhibited discontinuities in the Arrhenius plots, distinguishing the unfolding and aggregation phases of the denaturation process, whereas such a discrimination could not be obtained by differential scanning calorimetry analyses. The proposed method is accurate, fast, simple and sensitive enough to detect changes in the protein heat denaturation even at short temperature-time intervals.

Aqueous foams stabilized by chitin nanocrystals.

The aim of the present study was to explore the potential use of chitin nanocrystals, as colloidal rod-like particles, to stabilize aqueous foams. Chitin nanocrystals (ChN) were prepared by acid hydrolysis of crude chitin and foams were generated mainly by sonicating the respective dispersions. The foamability of the chitin nanocrystals was evaluated and the resulting foams were assessed for their stability, in terms of foam volume reduction and serum release patterns, during storage. Additionally, the samples were studied with light scattering and optical microscopy in order to explore the bubble size distribution and morphology of the foam. Nanocrystal concentration and charge density was varied to alter the packing of the crystals at the interface. At low concentrations of ChNs, foams were stable against coalescence and disproportionation for a period of three hours, whereas at higher concentrations, the foams were stable for several days. The enhanced stability of foams prepared with ChNs, compared to surfactant-stabilized foams, can be mainly attributed to the irreversible adsorption of the ChNs at the air-water interface, thereby providing Pickering stabilization. Both foam volume and stability of the foam were increased with an increase in ChNs concentration, and at pH values around the chitin's pKa (pH 7.0). Under these conditions, the ChNs show minimal electrostatic repulsion and therefore a higher packing of the nanocrystals is promoted. Moreover, decreased electrostatic repulsion enhances network formation between the ChNs in the aqueous films, thereby providing additional stability by gel formation. Overall, ChNs were proven to be effective in stabilizing foams, and may be useful in the design of Pickering-stabilized food grade foams.

Effect of soluble polysaccharides addition on rheological properties and microstructure of chitin nanocrystal aqueous dispersions.

Mixtures of chitin nanocrystal aqueous dispersions (at pH 3.0) with soluble polysaccharides of varying molecular features were examined rheologically and microscopically, under different conditions of biopolymer concentration, ionic strength, pH and temperature. The addition of non-adsorbing polysaccharides (guar gum, locust bean gum and xanthan) as well as oppositely charged (κ-carrageenan) to a chitin nanocrystal dispersion, resulted in a network formation and the gel strength increased with the chitin nanocrystal concentration. In contrast, the chitin nanocrystal - chitosan or - pullulan mixed dispersions did not show any network formation (tanδ>1) at the concentration range examined. An increase in ionic strength and pH also resulted in an enhanced elasticity of the chitin nanocrystal-guar gum dispersions. Furthermore, an increase in the elastic modulus, which was irreversible upon cooling, was observed upon heating the chitin nanocrystal-polysaccharide mixed dispersions.

In vitro lipid digestion of chitin nanocrystal stabilized o/w emulsions.

Chitin nanocrystals (ChN) have been shown to form stable Pickering emulsions. These oil-in-water emulsions were compared with conventional milk (whey protein isolate, WPI, and sodium caseinate, SCn) protein-stabilized emulsions in terms of their lipid digestion kinetics using an in vitro enzymatic protocol. The kinetics of fatty acid release were evaluated as well as the change in oil droplet size of the respective emulsions during lipid digestion. The interfacial pressure was measured by addition of the duodenal components using drop tensiometry and the electrical charge of the oil droplets was also assessed, in an attempt to relate the interfacial properties with the stability of the emulsions towards lipolysis. Lipid hydrolysis in the ChN-stabilized emulsion was appreciably slower and the plateau values of the total concentration of fatty acids released were much lower, compared to the WPI- and SCn-stabilized emulsions. Moreover, the ChN-stabilized emulsions were relatively stable to coalescence during lipid digestion, whereas the WPI- and SCn-stabilized emulsions exhibited a significant increase in their droplet size. On the other hand, no major differences were shown among the different emulsion samples in terms of their interfacial properties. The increased stability of the ChN-stabilized emulsions towards lipolysis could be attributed to several underlying mechanisms: (i) strong and irreversible adsorption of the chitin nanocrystals at the interface that might inhibit an extensive displacement of the solid particles by bile salts and lipase, (ii) network formation by the nanocrystals in the bulk (continuous) phase that may reduce lipid digestion kinetics, and (iii) the ability of chitin, and consequently of ChNs, to impair pancreatic lipase activity. The finding that ChNs can be used to impede lipid digestion may have important implications for the design and fabrication of structured emulsions with controlled lipid digestibility that could provide the basis for the development of novel products that may promote satiety, reduce caloric intake and combat obesity.

Using particle tracking to probe the local dynamics of barley ß-glucan solutions upon gelation.

The sol-gel transition of aqueous barley ß-glucan solutions which undergo gelation with ageing has been studied by conventional bulk rheology, phase contrast microscopy and particle tracking microrheology. Characterisation of the primary structure of the ß-glucan isolate was carried out by enzymic methods and HPLC. The Brownian diffusion of fluorescent microspheres (0.75 µm diameter, carboxylate-coated particles) was used to probe the spatial mechanical properties of the gelling systems at the scale of microns; the potential use of passive particle tracking to study biopolymer gelling systems that present spatial heterogeneities is thus explored. For the ß-glucan gels cured at 25°C both microrheology and bulk rheology revealed that with increasing the polysaccharide concentration the gelation time decreased, while the gelation rate and gel strength of the barley ß-glucan gels increased. The particle tracking method had higher sensitivity and could map molecular ordering and structural heterogeneities in the evolving polysaccharide network at a micro-level. That is, different size pores were generated upon ageing with regions of depleted or less amount of ß-glucan molecules. Furthermore, this method could detect changes in the fine structure of the system before such events can be registered by bulk rheological measurements; i.e. microheterogeneity and aggregation of ß-glucan chains were revealed by particle tracking at earlier temporal stages of the experiment.

Engineering interfacial properties by anionic surfactant-chitosan complexes to improve stability of oil-in-water emulsions.

Oil-in-water emulsions (10% w/w n-tetradecane) were prepared at pH = 5.7 by using, as surface active agents, electrostatically formed complexes of sodium stearoyl lactylate (SSL) at a concentration of 0.4% (w/w) and chitosan (CH) in a concentration range between 0 and 0.48% w/w. The use of complexes in emulsions with a low concentration of CH (<0.24% w/w) resulted in highly flocculated systems; instead, with increased level of CH, the emulsions had a smaller average droplet size and exhibited greater stability during storage. Emulsions stabilised by SSL/CH complexes showed non-Newtonian flow behavior with pronounced shear thinning. Among all formulations studied none showed a gel-like behavior since in all cases the G' (storage modulus) was lower that G'' (loss modulus). Adsorption kinetics of pure SSL and SSL/CH complexes to the oil/water interfaces were evaluated using an automated drop tensiometer (ADT). Even though complexation of SSL with CH resulted in a delay of the adsorption of the surface active species at the oil/water interface, the inclusion of the polysaccharide resulted in substantially improved interfacial properties as indicated by a significant increase of the dilatational modulus. Furthermore, the enhanced interfacial properties of the emulsion droplets resulted in improved stability against freeze-thaw cycling. The results of this study may facilitate the development of frozen food products such as desserts with an ameliorated stability and favorable sensorial characteristics.