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The increasing demand for tissue replacement has encouraged scientists worldwide to focus on developing new biofabrication technologies. Multimaterials/cells printed with stringent resolutions are necessary to address the high complexity of tissues. Advanced inkjet 3D printing can use multimaterials and attain high resolution and complexity of printed structures. However, a decisive yet limiting aspect of translational 3D bioprinting is selecting the befitting material to be used as bioink; there is a complete lack of cytoactive bioinks with adequate rheological, mechanical, and reactive properties. This work strives to achieve the right balance between resolution and cell support through methacrylamide functionalization of a psychrophilic gelatin and new fluorosurfactants used to engineer a photo-cross-linkable and immunoevasive bioink. The syntonized parameters following optimal formulation conditions allow proficient printability in a PolyJet 3D printer comparable in resolution to a commercial synthetic ink (â¼150 µm). The bioink formulation achieved the desired viability (â¼80%) and proliferation of co-printed cells while demonstrating in vivo immune tolerance of printed structures. The practical usage of existing high-resolution 3D printing systems using a novel bioink is shown here, allowing 3D bioprinted structures with potentially unprecedented complexity.
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Bioimpresión , Bioimpresión/métodos , Impresión Tridimensional , Gelatina/química , Reología , Andamios del Tejido/química , Ingeniería de Tejidos/métodosRESUMEN
Ischemic stroke imparts elevated risk of heart failure though the underlying mechanisms remain poorly described. We aimed to characterize the influence of cerebral ischemic injury on cardiac function using multimodality molecular imaging to investigate brain and cardiac morphology and tissue inflammation in two mouse models of variable stroke severity. Transient middle cerebral artery occlusion (MCAo) generated extensive stroke damage (56.31 ± 40.39 mm3). Positron emission tomography imaging of inflammation targeting the mitochondrial translocator protein (TSPO) revealed localized neuroinflammation at 7 days after stroke compared to sham (3.8 ± 0.8 vs 2.6 ± 0.7 %ID/g max, p < 0.001). By contrast, parenchyma topical application of vasoconstrictor endothelin-1 did not generate significant stroke damage or neuroinflammatory cell activity. MCAo evoked a modest reduction in left ventricle ejection fraction at both 1 weeks and 3 weeks after stroke (LVEF at 3 weeks: 54.3 ± 5.7 vs 66.1 ± 3.5%, p < 0.001). This contractile impairment was paralleled by elevated cardiac TSPO PET signal compared to sham (8.6 ± 2.4 vs 5.8 ± 0.7%ID/g, p = 0.022), but was independent of leukocyte infiltration defined by flow cytometry. Stroke size correlated with severity of cardiac dysfunction (r = 0.590, p = 0.008). Statistical parametric mapping identified a direct association between neuroinflammation at 7 days in a cluster of voxels including the insular cortex and reduced ejection fraction (ρ = - 0.396, p = 0.027). Suppression of microglia led to lower TSPO signal at 7 days which correlated with spared late cardiac function after MCAo (r = - 0.759, p = 0.029). Regional neuroinflammation early after cerebral ischemia influences subsequent cardiac dysfunction. Total body TSPO PET enables monitoring of neuroinflammation, providing insights into brain-heart inter-organ communication and may guide therapeutic intervention to spare cardiac function post-stroke.
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Isquemia Encefálica , Cardiopatías , Accidente Cerebrovascular , Animales , Ratones , Encéfalo/diagnóstico por imagen , Encéfalo/metabolismo , Isquemia Encefálica/metabolismo , Endotelina-1/metabolismo , Cardiopatías/metabolismo , Infarto de la Arteria Cerebral Media/complicaciones , Infarto de la Arteria Cerebral Media/diagnóstico por imagen , Inflamación/metabolismo , Imagen Molecular/métodos , Tomografía de Emisión de Positrones/métodos , Accidente Cerebrovascular/metabolismo , VasoconstrictoresRESUMEN
Status epilepticus (SE) is a clinical emergency with high mortality. SE can trigger neuronal death or injury and alteration of neuronal networks resulting in long-term cognitive decline or epilepsy. Among the multiple factors contributing to this damage, imbalance between oxygen and glucose requirements and brain perfusion during SE has been proposed. Herein, we aimed to quantify by neuroimaging the spatiotemporal course of brain perfusion during and after lithium-pilocarpine-induced SE in rats. To this purpose, animals underwent 99mTc-HMPAO SPECT imaging at different time points during and after SE using a small animal SPECT/CT system. 99mTc-HMPAO regional uptake was normalized to the injected dose. In addition, voxel-based statistical parametric mapping was performed. SPECT imaging showed an increase of cortical perfusion before clinical seizure activity onset followed by regional hypo-perfusion starting with the first convulsive seizure and during SE. Twenty-four hours after SE, brain 99mTc-HMPAO uptake was widely decreased. Finally, chronic epileptic animals showed regionally decreased perfusion affecting hippocampus and cortical sub-regions. Despite elevated energy and oxygen requirements, brain hypo-perfusion is present during SE. Our results suggest that insufficient compensation of required blood flow might contribute to neuronal damage and neuroinflammation, and ultimately to chronic epilepsy generated by SE.
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Estado Epiléptico , Tomografía Computarizada de Emisión de Fotón Único , Animales , Encéfalo/irrigación sanguínea , Encéfalo/diagnóstico por imagen , Circulación Cerebrovascular/fisiología , Neuroimagen , Ratas , Estado Epiléptico/inducido químicamente , Estado Epiléptico/diagnóstico por imagen , Exametazima de Tecnecio Tc 99m , Tomografía Computarizada de Emisión de Fotón Único/métodosRESUMEN
Scleroglucan, a neutral ß(1-3) glucan with ß(1-6) glucan branches every third residue, is being considered as an alternative rod-like, shear thinning high molecular weight ß-glucan based polysaccharide to xanthan gum for the management of patients with oropharyngeal dysphagia. It is therefore important to understand more fully its hydrodynamic properties in solution, in particular heterogeneity, molecular weight distribution and its behaviour in the presence of mucin glycoproteins. A commercially purified scleroglucan preparation produced by fermentation of the filamentous fungus Sclerotium rolfsii was analysed in deionised distilled water with 0.02% added azide. Sedimentation velocity in the analytical ultracentrifuge showed the scleroglucan preparation to be unimodal at concentrations >0.75â¯mg/ml which resolved into two components at lower concentration and with partial reversibility between the components. Sedimentation coefficient versus concentration plots showed significant hydrodynamic non-ideality. Self-association behaviour was confirmed by sedimentation equilibrium experiments with molecular weights between ~3â¯×â¯106â¯g/mol to ~5â¯×â¯106â¯g/mol after correcting for thermodynamic non-ideality. SEC-MALS-viscosity experiments showed a transition between a rod-shape at lower molar masses to a more flexible structure at higher masses consistent with previous observations. Sedimentation velocity experiments also showed no evidence for potentially problematic interactions with submaxillary mucin.
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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.
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PURPOSE: Investigate the extended release behaviour of compacts containing mixtures of hydrophilic HPMC and PEO in hydrating media of differing ionic strengths. METHODS: The extended release behaviour of various HPMC:PEO compacts was investigated using dissolution testing, confocal microscopy and magnetic resonance imaging, with respect to polymer ratio and ionic strength of the hydrating media. RESULTS: Increasing HPMC content gave longer extended release times, but a greater sensitivity to high ionic dissolution environments. Increasing PEO content reduced this sensitivity. The addition of PEO to a predominantly HPMC matrix reduced release rate sensitivity to high ionic environments. Confocal microscopy of early gel layer development showed the two polymers appeared to contribute independently to gel layer structure whilst together forming a coherent and effective diffusion barrier. There was some evidence that poorly swollen HPMC particles added a tortuosity barrier to the gel layer in high ionic strength environments, resulting in prolonged extended release. MRI provides unique, non-invasive spatially resolved information from within the HPMC:PEO compacts that furthers our understanding of USP 1 and USP 4 dissolution data. CONCLUSIONS: Confocal microscopy and MRI data show that combinations of HPMC and PEO have advantageous extended release properties, in comparison with matrices containing a single polymer.
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Preparaciones de Acción Retardada/química , Derivados de la Hipromelosa/química , Iones/química , Polietilenglicoles/química , Interacciones Hidrofóbicas e Hidrofílicas , Concentración Osmolar , Polímeros/química , Sensibilidad y Especificidad , SolubilidadRESUMEN
Sodium (salt) was encapsulated within the inner water phase of w1/o/w2 food emulsions externally stabilised by starch particles with the ultimate aim of enhancing saltiness perception. The physical properties of the starch particles were modified by octenyl succinic anhydride (OSA) treatment (0-3%) to vary the degree of hydrophobicity of the emulsifying starch. During oral processing native salivary amylase hydrolysed the starch and destabilised the o/w emulsion releasing the inner w/o phase and subsequently sodium into the oral cavity, resulting in a salty taste. Whilst increasing OSA treatment levels increased the stability of the emulsion, intermediate or low levels of starch modification resulted in enhanced saltiness. It is therefore proposed that 1.5% OSA modified starch is optimal for sodium delivery and 2% OSA modified starch is optimal for sodium delivery in systems that require greater process stability. It is also shown that sodium release was further enhanced by oral processing and was positively correlated with native amylase activity. The results demonstrate a promising new approach for the reduction of salt or sugar in emulsion based foods.
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A particulate form of xanthan gum was prepared by extrusion cooking. The temperature dependence of the viscosity of this form shows similarities to starch with an increase in viscosity to a maximum with increasing temperature as a result of the swelling of the particles. The rheology and mixing behaviour with water of the particulate and conventional molecular forms of xanthan were compared with a modified starch. The particulate xanthan products mixed rapidly with water in a similar way to ungelatinised starch, whereas conventional molecular xanthan systems mixed poorly. Using an experienced sensory panel, model tomato products thickened with the three systems were compared at equal shear viscosities. The panel could not discriminate between the tomato flavour of the three products, but found that the xanthan products were perceived as being significantly thicker. These observations were consistent with previous work. Salt perception for both xanthan products was poorer than for the starch thickened systems. A hypothesis to explain why xanthan does not fit into the previously postulated link between mixing and perception is presented.
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The objective of the work reported in this paper is to determine if saliva addition has an effect on the rheology of xanthan gum solutions. The reasons for the interest was that it has been previously reported that flavour release from high viscosity xanthan thickened foods is not reduced in the same way as foods thickened by other hydrocolloids at comparable viscosities. It was previously postulated that this could be due to an interaction between saliva and xanthan that could change the microstructure and rheology of xanthan solutions. In this work the effect of saliva on the rheology of CMC and xanthan solutions was compared. Solutions of molecularly dissolved xanthan gum and CMC mixed with water or human whole saliva at a ratio of 5:1 showed little impact of the presence of saliva on steady shear or dynamic viscosity for the two hydrocolloids. In filament thinning experiments saliva addition significantly increased filament break-up time for xanthan gum while it had little effect on the break-up time of the CMC filament. Also, filament thinning appeared a lot less even and was not as reproducible in the case of xanthan gum. Addition of CMC and hydroxypropyl methylcellulose (HPMC) to xanthan gum solutions showed a similar increase in break-up time to saliva, but to see this effect the viscosity of the added CMC or HPMC solution had to be very much higher than the viscosity of saliva. The results are discussed in the context of the structure of xanthan gum and the reported extensional rheology of saliva.
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The typically low solubility and gelation capacity of plant proteins can impose challenges in the design of high-quality plant-based foods. The acid used during the precipitation step of plant protein isolate extraction can influence protein functionality. Here, acetic acid and citric acid were used to extract quinoa protein isolate (QPI) from quinoa flour, as these acids are more kosmotropic than the commonly used HCl, promoting the stabilisation of the native protein structure. While proximate analysis showed that total protein was similar for the three isolates, precipitation with kosmotropic acids increased soluble protein, which correlated positively with gel strength. Microstructure analysis revealed that these gels contained a less porous protein network with lipid droplet inclusions. This study shows that the choice of precipitation acid offers an opportunity to tailor the properties of quinoa protein isolate for application, a strategy that is likely applicable to other plant protein isolates.
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Chenopodium quinoa , Proteínas de Plantas , Chenopodium quinoa/química , Proteínas de Plantas/química , Proteínas de Plantas/aislamiento & purificación , Solubilidad , Ácido Cítrico/química , Ácido Acético/química , Precipitación Química , Harina/análisisRESUMEN
Background: The electrical cochlear implant (eCI) partially restores hearing in individuals affected by profound hearing impairment (HI) or deafness. However, the limited resolution of sound frequency coding with eCIs limits hearing in daily situations such as group conversations. Current research promises future improvements in hearing restoration which may involve gene therapy and optical stimulation of the auditory nerve, using optogenetics. Prior to the potential clinical translation of these technologies, it is critical that patients are engaged in order to align future research agendas and technological advancements with their needs. Methods: Here, we performed a survey study with hearing impaired, using an eCI as a means of hearing rehabilitation. We distributed a questionnaire to 180 adult patients from the University Medical Center Göttingen's Department of Otolaryngology who were actively using an eCI for 6 months or more during the time of the survey period. Questions revolved around patients needs, and willingness to accept hypothetical risks or drawbacks associated with an optical CI (oCI). Results: Eighty-one participants responded to the questionnaire; 68% were greater than 60 years of age and 26% had bilateral eCIs. Participants expressed a need for improving the performance beyond that experienced with their current eCI. Primarily, they desired improved speech comprehension in background noise, greater ability to appreciate music, and more natural sound impression. They expressed a willingness for engaging with new technologies for improved hearing restoration. Notably, participants were least concerned about hypothetically receiving a gene therapy necessary for the oCI implant; but expressed greater reluctance to hypothetically receiving an implant that had yet to be evaluated in a human clinical trial. Conclusion: This work provides a preliminary step in engaging patients in the development of a new technology that has the potential to address the limitations of electrical hearing rehabilitation.
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Cochlear implants (CIs) restore activity in the deafened auditory system via electrical stimulation of the auditory nerve. As the spread of electric current in biological tissues is rather broad, the spectral information provided by electrical CIs is limited. Optogenetic stimulation of the auditory nerve has been suggested for artificial sound coding with improved spectral selectivity, as light can be conveniently confined in space. Yet, the foundations for optogenetic sound coding strategies remain to be established. Here, we parametrized stimulus-response-relationships of the auditory pathway in gerbils for optogenetic stimulation. Upon activation of the auditory pathway by waveguide-based optogenetic stimulation of the spiral ganglion, we recorded neuronal activity of the auditory midbrain, in which neural representations of spectral, temporal, and intensity information can be found. Screening a wide range of optical stimuli and taking the properties of optical CI emitters into account, we aimed to optimize stimulus paradigms for potent and energy-efficient activation of the auditory pathway. We report that efficient optogenetic coding builds on neural integration of millisecond stimuli built from microsecond light pulses, which optimally accommodate power-efficient laser diode operation. Moreover, we performed an activity-level-dependent comparison of optogenetic and acoustic stimulation in order to estimate the dynamic range and the maximal stimulation intensity amenable to single channel optogenetic sound encoding, and indicate that it complies well with speech comprehension in a typical conversation (65 dB). Our results provide a first framework for the development of coding strategies for future optogenetic hearing restoration.
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Implantación Coclear , Implantes Cocleares , Vías Auditivas/fisiología , Optogenética/métodos , Mesencéfalo , Estimulación Acústica , Estimulación EléctricaRESUMEN
Lignin is the second most abundant natural polymer after cellulose, and valorisation of lignin-rich streams has attracted increasing attention recently. This paper presents a novel and sustainable method to recover lignin from Cocoa Bean Shells (CBS) using Deep Eutectic Solvents (DES) and microwaves. A DES containing p-toluenesulfonic acid, choline chloride and glycerol (2:1:1 M ratio) was selected based on its dielectric properties. Under 200 W microwave power, the optimum yield of 95.5 % lignin was achieved at 130 °C and 30 min. DES-extracted lignin exhibited unique structural characteristics including larger particle sizes (242.5 µm D50 size), structural diversity (410.4 µm D90-D10 size) and H/G sub-unit ratio (71.9 %) compared with commercial Kraft lignin (77.2 µm, 157.9 µm and 0.1 % respectively), indicating the potential of DES in the modification and upgrading of lignin for novel value-added products.
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Lignina , Microondas , Lignina/química , Disolventes Eutécticos Profundos , Solventes/química , Celulosa , BiomasaRESUMEN
Chloroplasts are abundant organelles in a diverse range of plant materials; they are predominantly composed of multicomponent thylakoid membranes which are lipid and protein rich. Intact or unravelled thylakoid membranes should, in principle, have interfacial activity, but little has been published on their activity in oil-in-water systems, and nothing on their performance on an oil continuous system. In this work different physical methods were used to produce a range of chloroplast/thylakoid suspensions with varying degrees of membrane integrity. Transmission electron microscopy revealed that pressure homogenisation led to the greatest extent of membrane and organelle disruption compared to less energy intensive preparation methods The ability of the derived materials to modulate the flow behaviour of a chocolate model system (65% (w/w) sugar/ sunflower oil (natural amphiphiles removed) suspension) was investigated by acquiring rheological parameters. All chloroplast/thylakoid preparations reduced yield stress, apparent viscosity, tangent flow point and cross over point in a concentration-dependent fashion, although not as significantly as polyglycerol polyricinoleate applied at a commercially relevant concentration in the same chocolate model system. Confocal laser scanning microscopy confirmed presence of the alternative flow enhancer material at the sugar surfaces. This research reveals that low-energy processing methods that do not extensively disrupt thylakoid membranes are applicable to generating materials with marked capacity to affect the flow behaviour of a chocolate model system. In conclusion, chloroplast/thylakoid materials hold strong potential as natural alternatives to synthetic rheology modifiers for lipid-based systems such as PGPR.
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Cacao , Tilacoides , Cloroplastos , Ácidos Ricinoleicos , AzúcaresRESUMEN
Consumers are increasingly looking for new plant-based alternatives to substitute animal proteins in their diets but for some applications it can be difficult to achieve the desired product microstructure using only plant proteins. One approach to facilitate structuring is to mix these plant-based ingredients with a polysaccharide. Here, the phase behaviour and microstructure of quinoa protein isolate (QPI) in mixture with maltodextrin (MD) of two dextrose equivalents (DE 7 and 2) were investigated. The binodals of both QPI-MD phase diagrams showed an atypical shape, where the concentration of MD in the QPI-rich phase and of QPI in the MD-rich phase increased with overall biopolymer concentration. Molecular weight distribution and microstructure analyses revealed that both maltodextrins fractionated between the phases and were probably entrapped within the volume-spanning protein network in the QPI-rich phase, indicating a depletion flocculation mechanism of phase separation. The pre-heating of QPI and the removal of salt from the systems resulted in similarly atypical phase diagrams. The approach presented contributes to our understanding of the phase behaviour of mixtures between plant proteins and polysaccharides, while the results suggest that the formulation of plant-based products of predictable properties may be more challenging than anticipated.
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Chenopodium quinoa , Animales , Polisacáridos , Proteínas de Plantas , Peso MolecularRESUMEN
Hearing impairment, the most prevalent sensory deficit, affects more than 466 million people worldwide (WHO). We presently lack causative treatment for the most common form, sensorineural hearing impairment; hearing aids and cochlear implants (CI) remain the only means of hearing restoration. We engaged with CI users to learn about their expectations and their willingness to collaborate with health care professionals on establishing novel therapies. We summarize upcoming CI innovations, gene therapies, and regenerative approaches and evaluate the chances for clinical translation of these novel strategies. We conclude that there remains an unmet medical need for improving hearing restoration and that we are likely to witness the clinical translation of gene therapy and major CI innovations within this decade.
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Implantación Coclear , Implantes Cocleares , Audífonos , Pérdida Auditiva Sensorineural , Pérdida Auditiva , Audición , Pérdida Auditiva/genética , Pérdida Auditiva/terapia , Pérdida Auditiva Sensorineural/genética , Pérdida Auditiva Sensorineural/terapia , HumanosRESUMEN
Improved hearing restoration by cochlear implants (CI) is expected by optical cochlear implants (oCI) exciting optogenetically modified spiral ganglion neurons (SGNs) via an optical pulse generated outside the cochlea. The pulse is guided to the SGNs inside the cochlea via flexible polymer-based waveguide probes. The fabrication of these waveguide probes is realized by using 6" wafer-level micromachining processes, including lithography processes such as spin-coating cladding layers and a waveguide layer in between and etch processes for structuring the waveguide layer. Further adhesion layers and metal layers for laser diode (LD) bonding and light-outcoupling structures are also integrated in this waveguide process flow. Optical microscope and SEM images revealed that the majority of the waveguides are sufficiently smooth to guide light with low intensity loss. By coupling light into the waveguides and detecting the outcoupled light from the waveguide, we distinguished intensity losses caused by bending the waveguide and outcoupling. The probes were used in first modules called single-beam guides (SBGs) based on a waveguide probe, a ball lens and an LD. Finally, these SBGs were tested in animal models for proof-of-concept implantation experiments.
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Equal parts of sugar beet pectin and sodium caseinate were interacted through electrostatic attraction, enzymatic crosslinking, and the Maillard reaction to prepare three oil-in-water emulsifier systems. Oil-in-water emulsions (10%) were processed via high shear overhead mixing at the natural pH of the emulsifier systems, followed by pH adjustment to pH 4.5 and pH 7. The emulsions were stable against coalescence, except for a slight increase in the mean droplet size for the enzymatic cross-liked emulsion at pH 4.5 over a 14-day storage period. This emulsion also showed the lowest absolute zeta (ζ)-potential value of near 30 mV. The Maillard interaction emulsifier system resulted in larger droplet sizes compared to the other two emulsifier systems. Small deformation oscillatory shear rheology assessment of the emulsion cream phases revealed an impact of the emulsifier system design at pH 4.5.
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This study concerns the preparation and functionality testing of a new class of Pickering particles for food emulsion stabilization: colloidal lignin-rich particles (CLRPs) derived from ethanol-soluble extract of cocoa shell. A further goal was to achieve Pickering functionality without the need to add co-emulsifying surfactants during emulsion processing. Cocoa shell is a co-product of the food manufacturing industry. As such it is anticipated that the particles would be accepted as a natural food ingredient, provided no harmful solvents are used in any step of their processing. The cocoa shell particles were milled, dispersed in water and exposed to 250 °C for 1 h in a stainless-steel tubular reactor followed by ethanol extraction to obtain a lignin-rich extract (46% (w/w) lignin with the remainder predominantly lipids). CLRPs were then fabricated by the precipitation of ethanol-dissolved extract into water (antisolvent). By employing an agitated process and droplet dosing into a non-agitated process, four particle suspensions of a range of submicron diameters were obtained. All particle suspensions contained the same mass fraction of extract and were surface active, with surface tension decreasing with increasing particle size. The smallest particles were obtained when lipids were removed from the extract prior to particle processing. In contrast to the other four particle suspensions, this one failed to stabilize a 10% (w/w) sunflower oil-in-water emulsion. We hypothesize that the phospholipids indigenously present in these CLRP formulations are a critical component for Pickering functionality. It can be concluded that we have successfully introduced a new class of Pickering particles, fabricated from an industry co-product and anticipated to be food grade.
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Ageing provokes a plethora of molecular, cellular and physiological deteriorations, including heart failure, neurodegeneration, metabolic maladaptation, telomere attrition and hair loss. Interestingly, on the molecular level, the capacity to induce autophagy, a cellular recycling and cleaning process, declines with age across a large spectrum of model organisms and is thought to be responsible for a subset of age-induced changes. Here, we show that a 6-month administration of the natural autophagy inducer spermidine in the drinking water to aged mice is sufficient to significantly attenuate distinct age-associated phenotypes. These include modulation of brain glucose metabolism, suppression of distinct cardiac inflammation parameters, decreased number of pathological sights in kidney and liver and decrease of age-induced hair loss. Interestingly, spermidine-mediated age protection was associated with decreased telomere attrition, arguing in favour of a novel cellular mechanism behind the anti-ageing effects of spermidine administration.