An examination of the impact of unmelted, melted, and deconstructed cheese on lipid metabolism: a 6-week randomised trial.

Background: Evidence suggests cheese has a favourable or neutral effect on cardiometabolic health, compared to butter. To date, studies have only considered the cheese matrix in its unmelted form, while the effect of melted cheese remains unknown. Objective: To test the effect of 6-week daily consumption of ∼40 g dairy fat, eaten in either as unmelted cheese, melted cheese, or in a fully deconstructed form, on markers of metabolic health in overweight adults aged ≥50 years of age. Design: A 6-week randomised parallel intervention, where 162 participants (43.3% male) received ∼40 g of dairy fat per day, in 1 of 3 treatments: (A) 120 g full-fat Irish grass-fed cheddar cheese, eaten in unmelted form (n 58); (B) 120 g full-fat Irish grass-fed cheddar cheese eaten in melted form (n 53); or (C) the equivalent components; butter (49 g), calcium caseinate powder (30 g), and Ca supplement (CaCO3; 500 mg) (n 51). Results: There was no difference in weight, fasting glucose, or insulin between the groups post-intervention. Melted cheese, compared to unmelted cheese, increased total cholesterol (0.23 ± 0.79 mmol L-1vs. 0.02 ± 0.67 mmol L-1, P = 0.008) and triglyceride concentrations (0.17 ± 0.39 mmol L-1vs. 0.00 ± 0.42 mmol L-1, P = 0.016). Melted cheese increased total cholesterol concentrations by 0.20 ± 0.15 mmol L-1 and triglyceride concentrations by 0.17 ± 0.08 mmol L-1 compared to unmelted cheese. No significant differences were observed between the cheese forms for change in HDL, LDL or VLDL cholesterol. Conclusion: Compared to unmelted cheese, melted cheese was found to increase total cholesterol and triglyceride concentrations in middle-aged, overweight adults with no effect on weight or glycaemic control.

Enhancing the bioaccessibility of vitamin D using mixed micelles - An in vitro study.

Vitamin-D deficiency is a global issue and a food fortification strategy may reduce deficiency levels. Mixed micelles (MM) are crucial to vitamin-D absorption in vivo and may enhance vitamin-D food fortification. This study compared the ability of MM based delivery systems to oil-in-water emulsions to improve vitamin-D bioaccessibility in vitro. Vitamin-D loaded emulsions were formed with olive or coconut oil alone or with added l-α-phosphatidylcholine, as well as two MM based systems. Particle size throughout digestion, fatty acid release, and vitamin-D bioaccessibility were measured. After digestion, particles in the MM size range (∼6-10 nm) were observed for emulsions but not for MM based systems. The bioaccessibility of vitamin-D in olive and coconut emulsions was 75% and 78%, respectively, and âˆ¼ 90% with added l-α-phosphatidylcholine. Bioaccessibility for the MM alone was 93% and 90% when mixed with a protein/lactose base. Overall, MM show good potential as a delivery system for vitamin-D in vitro.

Improving vitamin D3 stability to environmental and processing stresses using mixed micelles.

Deficiency of vitamin-D is prevalent globally and can lead to negative health consequences. The fat-soluble nature of vitamin-D, coupled with its sensitivity to heat, light and oxygen limits its incorporation into foods. Mixed micelles (MM) have potential to enhance bioavailability of vitamin-D. This study explores the stability of MM to food processing regimes and their ability to protect vitamin-D. Subjecting MM to a range of shearing speeds (8,000-20,500 rpm) and to high pressure processing (600 MPa, 120sec) resulted in no change in MM size (4.1-4.5 nm). MM improved the retention of vitamin-D following exposure to UV-C light, near UV/visible light, and heat treatment. MM suspensions protected vitamin-D over a four week storage period at refrigeration or freezer conditions. Overall MM show potential to protect vitamin-D from degradation encountered in food processing and storage and may be beneficial as a mechanism to fortify foods with vitamin-D.

Vitamin D3 bioaccessibility: Influence of fatty acid chain length, salt concentration and l-α-phosphatidylcholine concentration on mixed micelle formation and delivery of vitamin D3.

Vitamin D (VD) is a fat-soluble vitamin with high deficiency levels evident globally. Bioaccessibility of VD is influenced by formation of mixed micelles (MM) during digestion. This study assessed the impact of fatty acid (FA) type, phospholipid concentration on MM formation and stability of MM to salts. MM formation occurred at NaCl and KCl concentrations ranging from 20 to 100 mM, when octanoic acid (C8) or stearic acid (C18) were used. MM hydrodynamic size increased with increasing l-α-phosphatidylcholine concentration (1.5-7.5 mM) for both C8 and C18, above which concentration MM did not form. FA chain length impacted MM with hydrodynamic size increasing from 3.8 nm for decanoic acid (C10) to 4.4 nm for C18. VD3 incorporation in MM was not influenced by the FA used (C10 or C18). Understanding stability and formation of MM and VD3 loading is an essential first step towards manipulating food structures for improving delivery of VD.

Incorporation of bioactive dairy hydrolysate influences the stability and digestion behaviour of milk protein stabilised emulsions.

The physical stability of emulsions containing bioactive ingredients is an important aspect of functional food development. This research investigated the effects of a bioactive dairy hydrolysate with anti-inflammatory effects on the properties of oil-in-water emulsions (23% rapeseed oil and 1.5% w/w protein). This was determined by monitoring the effects of various combinations of sodium caseinate (NaCas) and NaCas hydrolysate (NaCasH) (NaCas : NaCasH; 100 : 0, 40 : 60, 30 : 70, 20 : 80 and 0 : 100) on the physico-chemical characteristics (particle size distribution, microstructure, adsorption of protein to the interface, viscosity and creaming stability) of emulsions. Currently, there is growing interest in designing functional foods that modulate lipid digestion. Therefore, emulsion breakdown and subsequent release of free fatty acids (FFAs) from selected NaCasH stabilised emulsions (40 : 60 and 0 : 100) was monitored during in vitro gastrointestinal digestion and compared to the behaviour of emulsions stabilised by NaCas alone. Inclusion of NaCasH generally decreased the stability of the emulsions except when added at a NaCas : NaCasH ratio of 40 : 60 which resulted in emulsions with equivalent stability to the NaCas stabilised emulsions. Although the 40 : 60 combination provided an emulsion system as stable as NaCas, during simulated digestion, these emulsions demonstrated a slower rate of FFA release. This was attributed to the 40 : 60 stabilised emulsions having much larger flocculated lipid droplets than NaCas emulsions, which resulted in reduced surface area and fewer binding sites for lipase adsorption. Accordingly, the 40 : 60 emulsions were hydrolysed more slowly. Emulsions containing only NaCasH exhibited extensive coalescence prior to and during digestion and thus displayed the slowest release of FFA. The results suggest that including NaCasH in the emulsifier blend yields emulsions with modified digestibility and may form the basis of controlling the digestion and release of fat-soluble nutrients in formulated foods. However, further studies are required to optimise the stability of these emulsions before inclusion in such applications.

In vitro evaluation of chitosan copper chelate gels as a multimicronutrient feed additive for cattle.

BACKGROUND: Effective micronutrient supplementation strategies are critical to ensure optimal health and productivity in livestock. The objective of this study was to develop a copper and vitamin (multimicronutrient) delivery system based on chitosan gel beads, and test its suitability, in vitro, for use as a cattle feed additive. RESULTS: Chitosan was chelated with copper sulfate to produce millimetre-scale gel matrices (∼2 mm). The copper content was significantly increased (from 61 to 95 mg g by adjusting pH to alkaline conditions post bead formation. The beads could subsequently be loaded with the model vitamin riboflavin to levels as high as 324 µg g-1 beads. Restricted rehydration of the dried gel matrices in simulated rumen fluid led to a sustained release of riboflavin with no copper released in these neutral conditions for up to 24 h, demonstrating copper rumen bypass. Moreover, sustained release of the mineral was observed in abomasal conditions of pH 2 over a 3 h period. CONCLUSIONS: The matrices showed rumen bypass for copper yet supplied nutritionally relevant levels of the free mineral in abomasal conditions, as required for effective supplementation in cattle. The controlled-release properties demonstrated by the matrices indicate their potential as a multimicronutrient functional feed additive to enhance cattle nutrition and productivity. © 2018 Society of Chemical Industry.

Peptidomic screening of bitter and nonbitter casein hydrolysate fractions for insulinogenic peptides.

Sodium caseinate hydrolysates (NaCaH) contain biologically active peptides that can positively influence human health. However, their intense bitterness hinders their inclusion in food products. To our knowledge, no studies have investigated whether a correlation between bitterness and bioactivity exists in NaCaH, so it is not yet known what effect selective removal of bitterness has on NaCaH bioactivity. A deeper understanding of the physicochemical characteristics affecting both bitterness and bioactivity is therefore needed. The aim of this study was to use in silico analysis to elucidate the relationship between bitterness and bioactivity of the insulinogenic NaCaH. The NaCaH fractions were generated by membrane filtration and flash chromatography and were subsequently evaluated for bitterness by a sensory panel. In this present study, peptidomic and bioinformatic processing of these NaCaH fractions allowed for the identification of insulinogenic peptides as well as other literature-identified peptides in each of the fractions. The results showed that the most bitter fraction contained the highest abundance of insulinogenic peptides, whereas another bitter fraction contained the highest abundance of other literature-identified bioactive peptides exhibiting angiotensin-converting enzyme-inhibition activity. Although some bioactive peptides were identified in the least bitter fractions, the abundance of these peptides was very low. These observations show a correlation between bitter taste and bioactivity, highlighting potential complications in removing bitterness while maintaining bioactivity. However, as the most bitter fraction contained the highest abundance of insulinogenic peptides, there is potential for using a lower dose of this enriched bioactive fraction to exert health benefits. The second most bitter fraction contained a very low abundance of insulinogenic peptides and other bioactive peptides. Therefore, removal of this fraction could reduce the NaCaH product's bitterness without significantly altering overall bioactive potential.

Validation of a paper-disk approach to facilitate the sensory evaluation of bitterness in dairy protein hydrolysates from a newly developed food-grade fractionation system.

Casein-hydrolysates (NaCaH) are desirable functional ingredients, but their bitterness impedes usage in foods. This study sought to validate a paper-disk approach to help evaluate bitterness in NaCaHs and to develop a food-grade approach to separate a NaCaH into distinct fractions, which could be evaluated by a sensory panel. Membrane filtration generated <0.2-µm and <3-kDa permeates. Further fractionation of the <3-kDa permeate by flash-chromatography generated four fractions using ethanol (EtOH) concentrations of 5, 10, 30 and 50%. As some fractions were poorly soluble in water, the fractions were resolubilzed in EtOH and impregnated into paper-disks for sensory evaluation. Bitterness differences observed in the membrane fractions using this sensory evaluation approach reflected those observed for the same fractions presented as a liquid. The flash-chromatography fractions increased in bitterness with an increase in hydrophobicity, except for the 50% EtOH fraction which had little bitterness. Amino acid analysis of the fractions showed enrichment of different essential amino acids in both the bitter and less bitter fractions. Practical Applications: The developed food-grade fractionation system, allowed for a simple and reasonably scaled approach to separating a NaCaH, into physicochemically different fractions that could be evaluated by a sensory panel. The method of sensory evaluation used in this study, in which NaCaH samples are impregnated into paper-disks, provided potential solutions for issues such as sample insolubility and limited quantities of sample. As the impregnated paper-disk samples were dehydrated, their long storage life could also be suitable for sensory evaluations distributed by mail for large consumer studies. The research, in this study, allowed for a greater understanding of the physicochemical basis for bitterness in this NaCaH. As some essential amino acids were enriched in the less bitter fractions, selective removal of bitter fractions could allow for the incorporation of the less bitter NaCaH fractions into food products for added nutritional value, without negatively impacting sensory properties. There is potential for this approach to be applied to other food ingredients with undesirable tastes, such as polyphenols.

Monitoring the progression of calcium and protein solubilisation as affected by calcium chelators during small-scale manufacture of casein-based food matrices.

Calcium and protein solubilisation during small-scale manufacture of semi-solid casein-based food matrices was investigated and found to be very different in the presence or absence of calcium chelating salts. Calcium concentrations in the dispersed phase increased and calcium-ion activity (ACa++) decreased during manufacture of the matrices containing calcium chelating salts; with ∼23% of total calcium solubilised by the end of manufacture. In the absence of calcium chelating salts, these concentrations were significantly lower at equivalent processing times and remained unchanged as did ACa++, throughout manufacture. The protein content of the dispersed phase was low (≤3% of total protein), but was significantly higher for matrices containing calcium chelating salts. This study elucidates the critical role of calcium chelating salts in modulating casein hydration and dispersion and gives an indication of the levels of soluble calcium and protein required to allow matrix formation during manufacture of casein-based food structures e.g. processed and analogue cheese.

Altering the level of calcium changes the physical properties and digestibility of casein-based emulsion gels.

Casein-based emulsion gels prepared with different types of lipid (i.e. milk fat or rapeseed oil) were formulated with high (774 mg Ca per 100 g) or low (357 mg Ca per 100 g) calcium levels by blending acid and rennet casein. Their physicochemical characteristics (i.e. composition, texture, microstructure & water mobility) and in vitro digestibility were compared to conventionally formulated high-calcium (723 mg Ca per 100 g) emulsion gels made from rennet casein with calcium chelating salts (CCS). CCS-free, high-calcium emulsion gels were significantly (p ≤ 0.05) softer than those with low calcium levels (possibly due to their shorter manufacture time and higher pH) and showed the highest rates of disintegration during simulated gastric digestion. Despite having a higher moisture to protein ratio, the high-calcium emulsion gels containing CCS had broadly similar hardness values to those of high-calcium concentration prepared without CCS, but had higher cohesiveness. The high-calcium matrices containing CCS had quite a different microstructure and increased water mobility compared to those made without CCS and showed the slowest rate (p ≤ 0.05) of disintegration in the gastric environment. Gastric resistance was not affected by the type of lipid phase. Conversely, fatty acid release was similar for all emulsion gels prepared from milk fat, however, high-calcium emulsion gels (CCS-free) prepared from rapeseed oil showed higher lipolysis. Results suggest that food matrix physical properties can be modified to alter resistance to gastric degradation which may have consequences for the kinetics of nutrient release and delivery of bioactives sensitive to the gastric environment.

Modification of aftertaste with a menthol mouthwash reduces food wanting, liking, and ad libitum intake of potato crisps.

This research investigated the effect of modifying the aftertaste of potato crisps on (1) temporal sensory perception and (2) appetite using three mouthwash conditions (no mouthwash, a water mouthwash, and a menthol mouthwash). For the sensory study, 17 screened female subjects were trained on the Temporal Dominance of Sensations (TDS) methodology. Subjects undertook TDS to monitor all sensory attributes during the mastication of a 2 g crisp until swallowing (at 20s), then conducted the mouthwash, and then continued the TDS task to monitor aftertaste until 90s. For the appetite study, 36 subjects (18 male, 18 female) completed 100 mm Visual Analogue Scales (VAS) for desire, liking, hunger, and thirst, followed by an ad libitum eating task. For the VAS scales testing, subjects chewed and swallowed a 2 g crisp, and then immediately conducted the mouthwash before completing the VAS scales. For the ad libitum task, subjects were given 12 min to consume as many crisps as they desired on a plate (up to 50 g). Every three minutes they were required to conduct a mouthwash. TDS results showed that in comparison with no mouthwash, the water mouthwash significantly reduced aftertaste attributes such as savoury, salty, and fatty mouthcoating, and the menthol mouthwash significantly increased aftertaste attributes of cooling, minty, and tingly. The water mouthwash did not influence desire and liking of crisps, or hunger and thirst. The water mouthwash did not influence ad libitum intake of the crisps over a 12 min period. The menthol mouthwash significantly reduced desire and liking of the crisps, as well as hunger and thirst. Furthermore, the menthol mouthwash significantly reduced ad libitum crisp intake by 29% over the 12 min period.

Effects of calcium chelators on calcium distribution and protein solubility in rennet casein dispersions.

This study investigated the effects of calcium chelating salts on calcium-ion activity (ACa(++)), calcium distribution, and protein solubility in model CaCl2 solutions (50 mmol L(-1)) or rennet casein dispersions (15 g/100 g). Disodium phosphate and trisodium citrate at concentrations of 10 and 30 mmol L(-1) and at ratios of 1:0, 2:1, 1:1, 1:2 and 0:1 were added to both systems. The CaCl2 system, despite its simplicity, was a good indicator of chelating salt-calcium interactions in rennet casein dispersions. Adding trisodium citrate either alone or as part of a mixed chelating salt system resulted in high levels of dispersed "chelated" calcium; conversely, disodium phosphate addition resulted in lower levels, while the ACa(++) decreased with increasing concentration of both chelating salts. Neither chelating salt produced high levels of soluble protein. Thus calcium chelating salts may play a more subtle role in modulating hydration during manufacture of casein-based matrices than simply solubilising calcium or protein.

Cold-set whey protein microgels as pH modulated immobilisation matrices for charged bioactives.

The ability of cold-set whey protein microgels to function as pH-sensitive immobilisation matrices for bioactives was investigated. A pH dependent interaction was confirmed between the microgels and charged bioactives and this binding was impeded by the presence of competing ions in the solution, suggesting an electrostatic interaction. The use of a computer generated prediction model for the pH-dependent association of the microgels and further bioactives (including cationic and anionic peptides) was validated. The prediction model was efficient at determining the pH at which the maximum microgel-bioactive interaction occurred. This study highlights the capabilities of these food-grade whey based microgels as matrices that enable the immobilisation of a variety of bioactives by a charge interaction, and shows the potential for these matrices to function as smart delivery systems, in which uptake and release of bioactives is facilitated by environmental pH change.

An in vivo study examining the antiinflammatory effects of chamomile, meadowsweet, and willow bark in a novel functional beverage.

The antioxidant and antiinflammatory properties of polyphenols are well documented in vitro but there are few human studies. A herbal beverage composed of chamomile, meadowsweet, and willow bark (CMW) was developed and tested for its antiinflammatory effect in a cohort of healthy adults (n = 20) during a 4-week intervention. Subjects were randomised to either the treatment (TG) or placebo group (PG). The three herbs under study, which have been used in traditional and alternative medicine, were delivered in a berry extract matrix. This berry extract was used as a control in the experiment. The objective was to assess the herbs' effects on systemic inflammation and joint function by examining circulating cytokines and mechanical joint flexibility. Blood serum was analyzed for cytokines IL-1ß, IL-6, and TNFα. There was an average decrease of 21.7% IL-1ß in the treatment group, whereas the decrease seen in the placebo group was 3% but these were not statistically significant. Quartile analysis based on baseline production of TNFα demonstrated a decrease in the treatment group's IL-6 levels. This group showed improvements in mechanical joint function and pain upon movement of joints specific to the knee and lower back. Overall, no significant antiinflammatory effects were seen. The evidence is therefore inconclusive and further investigations are required using a larger cohort with some degree of elevated inflammatory activity.

Inhibition of proinflammatory biomarkers in THP1 macrophages by polyphenols derived from chamomile, meadowsweet and willow bark.

Antiinflammatory compounds in the diet can alleviate excessive inflammation, a factor in the pathogenesis of common diseases such as rheumatoid arthritis, atherosclerosis and diabetes. This study examined three European herbs, chamomile (Matricaria chamomilla), meadowsweet (Filipendula ulmaria L.) and willow bark (Salix alba L.), which have been traditionally used to treat inflammation and their potential for use as antiinflammatory agents. Aqueous herbal extracts and isolated polyphenolic compounds (apigenin, quercetin and salicylic acid, 0-100 µM) were incubated with THP1 macrophages, and interleukin (IL)-1ß, IL-6 and tumour necrosis factor-alpha (TNF-α) were measured. At concentrations of 10 µM, both apigenin and quercetin reduced IL-6 significantly ( p < 0.05). Apigenin at 10 µM and quercetin at 25 µM reduced TNF-α significantly ( p < 0.05). Amongst the herbal extracts, willow bark had the greatest antiinflammatory activity at reducing IL-6 and TNF-α production. This was followed by meadowsweet and then chamomile. The lowest effective antiinflammatory concentrations were noncytotoxic (MTT mitochondrial activity assay). The Comet assay, which was used to study the protective effect of the isolated phenols against oxidative damage, showed positive results for all three polyphenols. These are the first findings that demonstrate the antiinflammatory capacity of these herbal extracts.

Flavouring reduced fat high fibre cheese products with enzyme modified cheeses (EMCs).

Medium (13%) and low (2%) fat imitation cheeses (pH 6 or 5.5) were flavoured with 5% w/w EMC containing 16%, 28% or 47% total free fatty acids (low to high levels of hydrolysis, respectively) and were examined by a sensory panel. Aroma active short-chain free fatty acids were monitored using gas chromatographic techniques. Regardless of cheese pH or EMC composition, panellists ranked all medium-fat cheeses similarly. Low-fat cheeses flavoured (pH 6 or 5.5) with low or medium lipolysis EMC were described as 'well-balanced' and 'cheesy' and were significantly more preferred to cheeses containing high hydrolysis EMCs. Low-fat cheeses were least preferred of all cheeses because of 'very intense' bursts of off-flavours. Lower pH cheeses were softer and less melting. Higher fat levels in imitation cheese modulated a greater retention of fat-based flavour compounds and improved their release during consumption more than did lower fat levels.