Understanding the Molecular Conformation and Viscoelasticity of Low Sol-Gel Transition Temperature Gelatin Methacryloyl Suspensions.

For biomedical applications, gelatin is usually modified with methacryloyl groups to obtain gelatin methacryloyl (GelMA), which can be crosslinked by a radical reaction induced by low wavelength light to form mechanically stable hydrogels. The potential of GelMA hydrogels for tissue engineering has been well established, however, one of the main disadvantages of mammalian-origin gelatins is that their sol-gel transitions are close to room temperature, resulting in significant variations in viscosity that can be a problem for biofabrication applications. For these applications, cold-water fish-derived gelatins, such as salmon gelatin, are a good alternative due to their lower viscosity, viscoelastic and mechanical properties, as well as lower sol-gel transition temperatures, when compared with mammalian gelatins. However, information regarding GelMA (with special focus on salmon GelMA as a model for cold-water species) molecular conformation and the effect of pH prior to crosslinking, which is key for fabrication purposes since it will determine final hydrogel's structure, remains scarce. The aim of this work is to characterize salmon gelatin (SGel) and salmon methacryloyl gelatin (SGelMA) molecular configuration at two different acidic pHs (3.6 and 4.8) and to compare them to commercial porcine gelatin (PGel) and methacryloyl porcine gelatin (PGelMA), usually used for biomedical applications. Specifically, we evaluated gelatin and GelMA samples' molecular weight, isoelectric point (IEP), their molecular configuration by circular dichroism (CD), and determined their rheological and thermophysical properties. Results showed that functionalization affected gelatin molecular weight and IEP. Additionally, functionalization and pH affected gelatin molecular structure and rheological and thermal properties. Interestingly, the SGel and SGelMA molecular structure was more sensitive to pH changes, showing differences in gelation temperatures and triple helix formation than PGelMA. This work suggests that SGelMA presents high tunability as a biomaterial for biofabrication, highlighting the importance of a proper GelMA molecular configuration characterization prior to hydrogel fabrication.

Stabilisation of oil-in-water emulsions with non-chemical modified gelatinised starch.

In this research, stabilisation of oil-in-water emulsions with non-chemically modified gelatinised starch is presented. Thus far only octenyl succinic anhydride (OSA) modified gelatinised starch has been known to adsorb at emulsion droplet interfaces, acting as emulsifiers. Screening a range of commercially available food starches revealed that a non-waxy rice starch, a waxy rice starch and the waxy maize starch PRIMA600 showed oil-in-water emulsifying ability following gelatinisation. The microstructure of emulsions formulated with 20% oil and 1% starch was stable for at least 3 months. Thermal, crystallinity and molecular property analyses as well as amylose and protein content revealed no obvious link to this property. Nevertheless, this research has provided the food industry with exciting results for the formulation of clean label emulsions. Moreover, it presents a concept for oral release food emulsions with destabilisation via salivary amylase digestion of the stabilising starch emulsifier.

Stability of Lactobacillus rhamnosus GG incorporated in edible films: Impact of anionic biopolymers and whey protein concentrate.

The incorporation of probiotics and bioactive compounds, via plasticised thin-layered hydrocolloids, within food products has recently shown potential to functionalise and improve the health credentials of processed food. In this study, choice of polymer and the inclusion of whey protein isolate was evaluated for their ability to stabalise live probiotic organisms. Edible films based on low (LSA) and high (HSA) viscosity sodium alginate, low esterified amidated pectin (PEC), kappa-carrageenan/locust bean gum (κ-CAR/LBG) and gelatine (GEL) in the presence or absence of whey protein concentrate (WPC) were shown to be feasible carriers for the delivery of L. rhamnosus GG. Losses of L. rhamnosus GG throughout the drying process ranged from 0.87 to 3.06 log CFU/g for the systems without WPC, losses were significantly reduced to 0 to 1.17 log CFU/g in the presence of WPC. Storage stability (over 25d) of L. rhamnosus GG at both tested temperatures (4 and 25 °C), in descending order, was κ-CAR/LBG > HSA > GEL > LSA = PEC. In addition, supplementation of film forming agents with WPC led to a 1.8- to 6.5-fold increase in shelf-life at 4 °C (calculated on the WHO/FAO minimum requirements of 6 logCFU/g), and 1.6-4.3-fold increase at 25 °C. Furthermore probiotic films based on HSA/WPC and κ-CAR/LBG/WPC blends had both acceptable mechanical and barrier properties.

Compositional and physicochemical factors governing the viability of Lactobacillus rhamnosus GG embedded in starch-protein based edible films.

Probiotic incorporation in edible films and coatings has been shown recently to be an efficient strategy for the delivery of probiotics in foods. In the present work, the impact of the compositional, physicochemical and structural properties of binary starch-protein edible films on Lactobacillus rhamnosus GG viability and stability was evaluated. Native rice and corn starch, as well as bovine skin gelatine, sodium caseinate and soy protein concentrate were used for the fabrication of the probiotic edible films. Starch and protein type both impacted the structural, mechanical, optical and thermal properties of the films, and the process loss of L. rhamnosus GG during evaporation-dehydration was significantly lower in the presence of proteins (0.91-1.07 log CFU/g) compared to solely starch based systems (1.71 log CFU/g). A synergistic action between rice starch and proteins was detected when monitoring the viability of L. rhamnosus GG over four weeks at fridge and room temperature conditions. In particular, a 3- to 7-fold increase in the viability of L. rhamnosus GG was observed in the presence of proteins, with sodium caseinate - rice starch based films offering the most enhanced stability. The film's shelf-life (as calculated using the FAO/WHO (2011) basis of 6 log viable CFU/g) ranged between 27-96 and 15-24 days for systems stored at fridge or room temperature conditions respectively.

Headspace volatiles profiles of different spring varieties and a wild relative of wheat flour.

Volatile profiling was conducted on four wheat varieties Triticum aestivum cv. Chinese Spring (CS), Highbury (High), Paragon (Para), Pavon76 (Pav76), and one wild relative Triticum timopheevii (P95). Headspace solid-phase microextraction (SPME) combined with gas chromatography-mass spectrometry (GC-MS) was used to explore differences in flavor formation mechanisms in different flours before and after starch gelatinization. Solvent retention capacity (SRC) analysis revealed subtle differences in water absorption, gluten strength, and starch characteristics across wheat flour types. Rapid Visco Analysis (RVA) of whole wheat flour demonstrated significant variations in pasting properties among wheat varieties, with P95 exhibiting higher viscosities compared to CS and High potentially influenced by starch gelatinization, protein-starch interactions, and lipid content. Aroma contributions of P95 clustered positively in PCA plots, contrasting with the four main varieties, indicative of species-level differentiation. Furthermore, the study highlighted the roles of viscosity, protein structure, lipid content, and fatty acid composition in modulating the release and perception of volatile aroma compounds during heating. This study sheds light on how the distinct characteristics of wheat flour influence aroma profiles, revealing species-level differences and the pivotal role of physiochemical properties in shaping flavor development mechanisms.

Improvement of human skin cell growth by radiation-induced modifications of a Ge/Ch/Ha scaffold.

Gelatin-/chitosan-/hyaluronan-based biomaterials are used in tissue engineering as cell scaffolds. Three gamma radiation doses (1, 10 and 25 kGy) were applied to scaffolds for sterilization. Microstructural changes of the irradiated polymers were evaluated by using scanning electron microscopy (SEM) and differential scanning calorimetry (DSC). A dose of 25 kGy produced a rough microstructure with a reduction of the porosity (from 99 to 96 %) and pore size (from 160 to 123 µm). Radiation also modified the glass transition temperature between 31.2 and 42.1 °C (1 and 25 kGy respectively). Human skin cells cultivated on scaffolds irradiated with 10 and 25 kGy proliferated at 48 h and secreted transforming growth factor ß3 (TGF-ß3). Doses of 0 kGy (non-irradiated) or 1 kGy did not stimulate TGF-ß3 secretion or cell proliferation. The specific growth rate and lactate production increased proportionally to radiation dose. The use of an appropriate radiation dose improves the cell scaffold properties of biomaterials.

Effect of sweeteners and carbonation on aroma partitioning and release in beverage systems.

The effect of monosaccharides (glucose, fructose and galactose) and disaccharides (sucrose and lactose) at 10, 20 and 30 % w/v on the in-vitro aroma partitioning of C4 - C10 aldehydes and ethyl esters, as well as limonene (concentration of aroma compounds at 1 µg mL-1), was studied using atmospheric pressure chemical ionisation-mass spectrometry. An increase in sugar concentration from 0 to 30 % w/v resulted in a significant increase in partitioning under static headspace conditions for the majority of the compounds (p < 0.05), an effect generally not observed when 10 % w/v sucrose was substituted with low-calorie sweeteners (p > 0.05). The complexity of the system was increased to model a soft drink design - comprising water, sucrose (10, 20 and 30 % w/v), acid (0.15 % w/v), carbonation (∼7.2 g/L CO2) and aroma compounds representative of an apple style flavouring, namely ethyl butanoate and hexanal (10 µg mL-1 each). Although the addition of sucrose had no significant in-vivo effect, carbonation significantly decreased breath-by-breath (in-vivo) aroma delivery (p < 0.05). To understand the physical mechanisms behind aroma release from the beverage matrix, the effect of sucrose on the kinetics of the matrix components was explored. An increase in sucrose concentration from 0 to 30 % w/v resulted in a significant decrease in water activity (p < 0.05), which accounted for the significantly slower rate of self-diffusion of aroma compounds (p < 0.05), measured using diffusion-ordered spectroscopy-nuclear magnetic resonance spectroscopy. No significant effect of sucrose on carbon dioxide volume flux was found (p > 0.05).

Mushroom and cereal ß-D-glucan solid state NMR and FTIR datasets.

In the associated published article Kremmyda et al. (2021), the C1 region of the NMR spectra were examined in detail, as the C1 carbon was common to the crucial linkages in the glucan, and the most clearly separated region in the spectra. We provide the original measurement data here in topspin format, in order that different authors can examine the regions corresponding to the other carbons and in addition repeat our processing of the FID's using different parameters. Mushroom, Cereal, Reference and other samples, such as salts and salt hydrates have been measured. A series of different experiments on individual samples is also given. All of the samples and experiments available in the database are summarised in the Table below. We also provide an FTIR database in the form of an excel workbook with spectra in frequency/absorbence pairs.

Illuminating Host-Parasite Interaction at the Cellular and Subcellular Levels with Infrared Microspectroscopy.

Toxoplasma gondii (T. gondii) is an opportunistic protozoan that can cause brain infection and other serious health consequences in immuno-compromised individuals. This parasite has a remarkable ability to cross biological barriers and exploit the host cell microenvironment to support its own survival and growth. Recent advances in label-free spectroscopic imaging techniques have made it possible to study biological systems at a high spatial resolution. In this study, we used conventional Fourier-transform infrared (FTIR) microspectroscopy and synchrotron-based FTIR microspectroscopy to analyze the chemical changes that are associated with infection of human brain microvascular endothelial cells (hBMECs) by T. gondii (RH) tachyzoites. Both FTIR microspectroscopic methods showed utility in revealing the chemical alterations in the infected hBMECs. Using a ZnS hemisphere device, to increase the numerical aperture, and the synchrotron source to increase the brightness, we obtained spatially resolved spectra from within a single cell. The spectra extracted from the nucleus and cytosol containing the tachyzoites were clearly distinguished. RNA sequencing analysis of T. gondii-infected and uninfected hBMECs revealed significant changes in the expression of host cell genes and pathways in response to T. gondii infection. These FTIR spectroscopic and transcriptomic findings provide significant insight into the molecular changes that occur in hBMECs during T. gondii infection.

Compositional, thermal and water sorption properties of wheat flour: comparing wild-relatives v/s elite wheat varieties used in current plant breeding.

Plant breeders are focused on creating new varieties of wheat with enhanced yield potential without affecting the quality of wheat for food. In this study, four cultivated varieties of wheat, Triticium asetivum and two wild relatives of wheat; Triticum timopheevii and Triticum urartu were studied. The impact of starch and protein content on structural, thermal and water sorption properties of four main wheat cultivars in comparison to the wild relatives has been studied. Wild relatives of wheat required higher temperatures to initiate starch gelatinisation and amylose/lipid complex formation as compared to the four main wheat cultivars. The carbohydrate region in FTIR indicates structural differences (ordered and unordered starch) in the different varieties of wheat flour. The differentiation between wild relatives and main varieties is at the species level. Both wild relatives showed a noticeable difference in moisture sorption behaviour as compared with the 4 main cultivars, especially in terms of monolayer (M0) and the strength of water binding to the primary sites (C) values.

The detection, purity and structural properties of partially soluble mushroom and cereal ß-D-glucans: A solid-state NMR study.

ß-D-glucans are proposed to have many health benefits. It is therefore important to have methods which can distinguish these from other carbohydrates present in natural products, as well as giving glucan content and structural information. Correlations between features in the CP/MAS spectra of ß-D-glucans and enzyme assay determined ß-D-glucan content were generally found to be poor. The ß-D-glucan in dry and hydrated forms of the mushroom Ganoderma lucidum was investigated in detail by spectral peak fitting to the anomeric carbon C1 region in CP/MAS NMR spectra. Hydrated samples gave spectra with enhanced resolution and suggested that a clear distinction between ß-D-glucans and other carbohydrates could be possible in the anomeric carbon C1 region. Chemical shift values for a range of carbohydrate polymers, which can be found alongside ß-D-glucans, as well as the values for various linkages are given. Contamination by other carbohydrates and buffer salts is discussed.

Properties of the Extracellular Polymeric Substance Layer from Minimally Grown Planktonic Cells of Listeria monocytogenes.

The bacterium Listeria monocytogenes is a serious concern to food processing facilities because of its persistence. When liquid cultures of L. monocytogenes were prepared in defined media, it was noted that planktonic cells rapidly dropped out of suspension. Zeta potential and hydrophobicity assays found that the cells were more negatively charged (-22, -18, -10 mV in defined media D10, MCDB 202 and brain heart infusion [BHI] media, respectively) and were also more hydrophobic. A SEM analysis detected a capsular-like structure on the surface of cells grown in D10 media. A crude extract of the extracellular polymeric substance (EPS) was found to contain cell-associated proteins. The proteins were removed with pronase treatment. The remaining non-proteinaceous component was not stained by Coomassie blue dye and a further chemical analysis of the EPS did not detect significant amounts of sugars, DNA, polyglutamic acid or any other specific amino acid. When the purified EPS was subjected to attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy, the spectra obtained did not match the profile of any of the 12 reference compounds used. An x-ray diffraction (XRD) analysis showed that the EPS was amorphous and a nuclear magnetic resonance (NMR) analysis detected the presence of glycerol. An elemental energy dispersive x-ray (EDX) analysis showed traces of phosphorous as a major component. In conclusion, it is proposed that the non-proteinaceous component may be phospholipid in nature, possibly derived from the cell wall lipoteichoic acid.

The role of sodium chloride in the sensory and physico-chemical properties of sweet biscuits.

Salt is included in many foods which consumers do not regard as salty. This "hidden-salt" may offer functional benefits but is often overlooked in sodium reduction strategies. This study investigated its role in shortbread-like sweet biscuits (1.05 g NaCl/100 g). Sensory tests revealed significant flavour and texture differences after a salt reduction of 33% (0.86 g/ 100 g). This was explained by differences in the partitioning of hydrophobic aroma compounds into the headspace and a significant impact on structure. Texture analysis and X-ray-µCT measurements revealed a reduced hardness with larger and more air cells in salt-reduced biscuits. It is suggested that salt impacts on cereal proteins by altering their aggregation around flour particles and at bubble walls and that slower water loss occurs in salted matrices during baking. Hence, this study revealed the key properties significantly affected by salt reduction and proposes an explanation which will help to develop a targeted "hidden-salt" reduction strategy.

Comparison of the mechanical properties of cellulose and starch films.

Some effects of water at levels up to 25% (dry solids basis db) on regenerated cellulose films with a thickness of 100 mum were investigated by dynamic vapor sorption (DVS), X-ray diffraction, tensile testing, and proton-nuclear magnetic resonance (NMR). The sorption isotherm fitted by the D'Arcy and Watt model and the increase in NMR T(2) with water content suggest that a mobile water fraction appears at water contents above 10%db. Water absorption increased the crystallinity of cellulose films from 31% (dry) to 38% (25%db) and altered the dimensions of crystallites. Mechanical measurements on planar and notched samples at all the water contents used here showed ductile fracture behavior. Although the properties of water in these cellulose films are comparable to previously reported data on starch, cellulose films at low water content are much less brittle than starch. The reasons for this difference are explored.

The state of water and fat during the maturation of Cheddar cheese.

Cheddar cheese predicted to develop into different quality classes has been evaluated by time domain Nuclear Magnetic Resonance, Thermogravimetric analysis and quantitative sensory analysis. The water and fat proton signals in the transverse relaxation decay curves have been deconvoluted. Proton transverse relaxation values for both the water and fat fractions decrease and the relative %age of the proton peak area, predominantly from the fat increases over a 450-day ripening period. The thermodynamic free water percentage increases during maturation. Water and fat attributes can distinguish between Cheddar cheese batches after 56 days. Cheese batches which have lower transverse relaxation values for the water and fat proton fractions and a higher relative %age of the proton peak area predominantly from fat at 56 days, mature after 270 days to be more yellow, rubbery and smooth, have a less sour and lingering aftertaste and are also harder to form into a cheese ball.

Freshwater microplastic concentrations vary through both space and time.

Plastic pollution represents one of the most salient indicators of society's impact on the environment. The microplastic component of this is ubiquitous, however, microplastic studies are seldom representative of the locations they sample. Over 12 months we explored spatiotemporal variation in microplastic prevalence across a freshwater system and in atmospheric deposition within its catchment, in one of the most temporally comprehensive studies of microplastic pollution. Microplastics were quantified in low concentrations (max 0.4 particles L-1) at all freshwater sites, including upstream of urban areas, and on rivers that do not receive wastewater treatment plant effluent. Extrapolated microplastic abundances at each site varied by up to 8 orders of magnitude over the course of the sampling campaign, suggesting that microplastic surveys that do not account for temporal variability misrepresent microplastic prevalence. Whilst we do not wish to underplay the potential impacts of microplastic particles in the environment, we argue that microplastic pollution needs to be placed in a more critical context, including assessment of temporal variability, to appropriately inform legislators and consumers.

Impact of Neospora caninum Infection on the Bioenergetics and Transcriptome of Cerebrovascular Endothelial Cells.

In this work, the effects of the protozoan Neospora caninum on the bioenergetics, chemical composition, and elemental content of human brain microvascular endothelial cells (hBMECs) were investigated. We showed that N. caninum can impair cell mitochondrial (Mt) function and causes an arrest in host cell cycling at S and G2 phases. These adverse effects were also associated with altered expression of genes involved in Mt energy metabolism, suggesting Mt dysfunction caused by N. caninum infection. Fourier Transform Infrared (FTIR) spectroscopy analysis of hBMECs revealed alterations in the FTIR bands as a function of infection, where infected cells showed alterations in the absorption bands of lipid (2924 cm-1), amide I protein (1649 cm-1), amide II protein (1537 cm-1), nucleic acids and carbohydrates (1092 cm-1, 1047 cm-1, and 939 cm-1). By using quantitative synchrotron radiation X-ray fluorescence (µSR-XRF) imaging and quantification of the trace elements Zn, Cu and Fe, we detected an increase in the levels of Zn and Cu from 3 to 24 h post infection (hpi) in infected cells compared to control cells, but there were no changes in the level of Fe. We also used Affymetrix array technology to investigate the global alteration in gene expression of hBMECs and rat brain microvascular endothelial cells (rBMVECs) in response to N. caninum infection at 24 hpi. The result of transcriptome profiling identified differentially expressed genes involved mainly in immune response, lipid metabolism and apoptosis. These data further our understanding of the molecular events that shape the interaction between N. caninum and blood-brain-barrier endothelial cells.

Effect of moisture content on thermal and water absorption properties of microfibrillar cellulose with polymeric additives.

The aim of this study was 1) to investigate the influence of polymeric additives such as carboxyl methyl cellulose (CMC) and locust bean gum (LBG) added before and after homogenisation on the moisture uptake of microfibrillar cellulose (MFC) in the dry and semi-wet state; and 2) to further understand the thermally induced structural transitions of low moisture MFC in the presence of the polymeric additives. A higher moisture content in the highly dense MFC network maintains the fibrillated network structure, which is lost during the drying process resulting in MFC aggregates. The addition of polymeric additives results in the regaining of the structure upon redispersion of the dry material with CMC being more effective than LBG). Results also indicated that CMC has a high level of compatibility with MFC, whereas LBG appears to have limited distribution in the MFC dense microfibrillar network and probably exists as a separate phase when added after homogenisation, however co-processing of LBG and cellulose significantly changed this behaviour. The presence of low-temperature transitions in MFC/additives/water mixtures indicates the involvement of these semi-flexible polymeric additives in the formation of liquid crystals when added to MFC in low moisture environments (2% and 20% w/w). An insight is offered into the theory of surface interactions between MFC and polymeric additives, which prevents the agglomeration of microfibrils present in the highly fibrillated suspension upon drying.

Freshwater and airborne textile fibre populations are dominated by 'natural', not microplastic, fibres.

The potential role of natural textile fibres as environmental pollutants has been speculated upon by some environmental scientists, however, there is a general consensus that their biodegradability reduces their environmental threat. Whilst the risks that they pose remain poorly understood, their environmental prevalence has been noted in several recent microplastic pollution manuscripts. Here we highlight the extent to which natural textile fibres dominate fibre populations of upstream reaches of the River Trent, UK, as well as the atmospheric deposition within its catchment, over a twelve month microplastic sampling campaign. Across 223 samples, natural textile fibres represented 93.8% of the textile fibre population quantified. Moreover, though microplastic particles including synthetic fibres are known to be pervasive environmental pollutants, extruded textile fibres were absent from 82.8% of samples. Natural textile fibres were absent from just 9.7% of samples.

Interactions between microfibrillar cellulose and carboxymethyl cellulose in an aqueous suspension.

New microstructures with interesting, unique and stable textures, particularly relevant to food systems were created by redispersing Microfibrillar cellulose (MFC). This paper reports the interactions between microfibrillar cellulose and carboxymethyl cellulose (CMC) in redispersed aqueous suspensions, by using rheological measurements on variable ratios of MFC/CMC and correlating these with apparent water mobility as determined by time domain NMR. MFC is a network of cellulose fibrils produced by subjecting pure cellulose pulp to high-pressure mechanical homogenisation. A charged polymer such as CMC reduces the aggregation of microfibrillar/fibre bundles upon drying. Small amplitude oscillatory rheological analysis showed viscoelastic gel-like behaviour of suspensions which was independent of the CMC content in the MFC suspension. A viscous synergistic effect was observed when CMC was added to MFC before drying, leading to improved redispersibility of the suspension. Novel measurements using NMR relaxation suggested that the aggregated microfibrillar/fibre bundles normally dominate the relaxation times (T2). The dense microfibrillar network plays an important role in generating stable rheological properties and controlling the mobility of the polymer and hence the apparent mobility of the water in the suspensions.