Depletion of HP1α alters the mechanical properties of MCF7 nuclei.

Within the nucleus of the eukaryotic cell, DNA is partitioned into domains of highly condensed, transcriptionally silent heterochromatin and less condensed, transcriptionally active euchromatin. Heterochromatin protein 1α (HP1α) is an architectural protein that establishes and maintains heterochromatin, ensuring genome fidelity and nuclear integrity. Although the mechanical effects of changes in the relative amount of euchromatin and heterochromatin brought about by inhibiting chromatin-modifying enzymes have been studied previously, here we measure how the material properties of the nuclei are modified after the knockdown of HP1α. These studies were inspired by the observation that poorly invasive MCF7 breast cancer cells become more invasive after knockdown of HP1α expression and that, indeed, in many solid tumors the loss of HP1α correlates with the onset of tumor cell invasion. Atomic force microscopy (AFM), optical tweezers (OT), and techniques based on micropipette aspiration (MA) were each used to characterize the mechanical properties of nuclei extracted from HP1α knockdown or matched control MCF7 cells. Using AFM or OT to locally indent nuclei, those extracted from MCF7 HP1α knockdown cells were found to have apparent Young's moduli that were significantly lower than nuclei from MCF7 control cells, consistent with previous studies that assert heterochromatin plays a major role in governing the mechanical response in such experiments. In contrast, results from pipette-based techniques in the spirit of MA, in which the whole nuclei were deformed and aspirated into a conical pipette, showed considerably less variation between HP1α knockdown and control, consistent with previous studies reporting that it is predominantly the lamins in the nuclear envelope that determine the mechanical response to large whole-cell deformations. The differences in chromatin organization observed by various microscopy techniques between the MCF7 control and HP1α knockdown nuclei correlate well with the results of our measured mechanical responses and our hypotheses regarding their origin.

Complexes of ß-lactoglobulin and high methyl-esterified pectin as a one-shot delivery system for reinforcing oil/water interfaces.

Electrostatic complexation of negatively charged polysaccharides with ß-lactoglobulin (ß-lg) has been shown to bolster the protein films at oil/water interfaces thereby improving emulsion stability. However, recent sub-phase exchange experiments demonstrated that highly charged polysaccharides such as low methyl-esterified pectin are complementary only if sequentially introduced to a pre-formed interfacial ß-lg film. In this study, results of transient interfacial shear rheology show that, by using high-methylesterified pectins instead, complexes can be formed in pre-mixed solutions with ß-lg at pH 4 that can lead to reinforced protein films at dodecane/water interfaces. Using this one-shot adsorption of such complexes, pectins as well as short chain polysaccharides like homogalacturonan nearly doubled the steady state shear elastic moduli as compared to that of a pure ß-lg film. The lag times of film formation were established to be primarily decided by the charge density and pattern on the polysaccharide. Based on the results from mixed solutions of ß-lg monomers, it is proposed that the polysaccharide at pH 4 strengthens the resulting interfacial layer by concatenating adsorbed ß-lg molecules thereby establishing cross-links in the aqueous phase.

Lime and/or Phosphate Application Affects the Stability of Soil Organic Carbon: Evidence from Changes in Quantity and Chemistry of the Soil Water-Extractable Organic Matter.

The mechanisms by which lime and/or phosphate addition impacts the preservation of soil organic matter (OM) are poorly understood. We explored the changes in quantity and chemistry of water-extractable organic matter (WEOM) in the bulk soil and its heavy density fraction (>1.6 g/cm3) of an unmanaged C-rich volcanic soil caused by lime and/or phosphate application. The addition of lime or phosphate caused (i) a significant increase in the WEOM, along with a decrease in its C/N ratio and an increase in its aromaticity, and (ii) changes in the WEOM chemical composition, measured with pyrolysis-gas chromatography/mass spectrometry, this being most impacted by lime application. The combined effect of lime and phosphate addition on the quantity and chemistry of WEOM was larger than the effects of separate lime and phosphate additions. By comparing the response of the bulk soil and the heavy fraction, we infer that phosphate has a greater contribution to the destabilization of vulnerable particulate OM, while lime causes a comparable disruption in the particulate OM and that in the heavy fraction. These findings provide a mechanistic insight into the decreased OM stability after liming and/or P fertilizing Andosols. They have implications for designing climate-smart management practices for these soils.

Sustained-Release Injectable Hydrogel Formulations for Administration of Sodium Salicylate in Broiler Chickens.

We developed injectable hydrogels for the slow release of analgesic drugs in birds as an in vivo model of pharmacokinetics in wild avian species. Hydrogels loaded with sodium salicylate (NaSA) were injected subcutaneously in Ross broiler chickens. The hydrogels were made by dissolving sodium alginate and NaSA in water at 2 different concentrations (low, LALG; high, HALG) and then adding calcium chloride. In vitro drug release studies were performed by swelling the hydrogels in water and analyzing serial samples by ultraviolet-visible (UV-Vis) spectroscopy. Dried hydrogel films of the same formulations of the two alginate concentrations then were dissolved in sterile water for the in vivo pharmacokinetic study conducted in 18 chickens divided into 3 groups of 6 birds. Each of the 2 resultant NaSA hydrogel solutions were filtered with 0.2-µm syringe filters before injecting at a NaSA dose of 150 mg/kg SC in the respective LALG or HALG groups. The control group was injected SC with the same dose of NaSA dissolved in water. Pharmacokinetics parameters calculated by the compartmental and noncompartmental approaches were compared among the 3 groups by the Kruskal-Wallis test. Results of in vitro studies showed that both hydrogels released 80% of the drug during the first 3.5 hours. Results of the pharmacokinetic study indicated that NaSA concentrations remained above the minimum effective concentration (MEC) for analgesia in humans for 24 ± 8.9 (LALG) to 26 ± 4 (HALG) hours for the hydrogel formulations compared to 10 ± 5.6 hours for the aqueous formulation. These hydrogel formulations may have potential in providing long-term analgesia in avian species, but need further evaluation with pharmacodynamic or pharmacokinetic/pharmacodynamic modeling studies.

The role of porous nanostructure in controlling lipase-mediated digestion of lipid loaded into silica particles.

The rate and extent of lipolysis, the breakdown of fat into molecules that can be absorbed into the bloodstream, depend on the interfacial composition and structure of lipid (fat) particles. A novel method for controlling the interfacial properties is to load the lipid into porous colloidal particles. We report on the role of pore nanostructure and surface coverage in controlling the digestion kinetics of medium-chain and long-chain triglycerides loaded into porous silica powders of different particle size, porosity, and hydrophobicity/hydrophilicity. An in vitro lipolysis model was used to measure digestion kinetics of lipid by pancreatic lipase, a digestive enzyme. The rate and extent of lipid digestion were significantly enhanced when a partial monolayer of lipid was loaded in porous hydrophilic silica particles compared to a submicrometer lipid-in-water emulsion or a coarse emulsion. The inhibitory effect of digestion products was clearly evident for digestion from a submicrometer emulsion and coarse emulsion. This effect was minimal, however, in the two silica-lipid systems. Lipase action was inhibited for lipid loaded in the hydrophobic silica and considered due to the orientation of lipase adsorption on the methylated silica surface. Thus, hydrophilic silica promotes enhanced digestion kinetics, whereas hydrophobic silica exerts an inhibitory effect on hydrolysis. Evaluation of digestion kinetics enabled the mechanism for enhanced rate of lipolysis in silica-lipid systems to be derived and detailed. These investigations provide valuable insights for the optimization of smart food microparticles and lipid-based drug delivery systems based on lipid excipients and porous nanoparticles.

Coalescence in concentrated Pickering emulsions under shear.

We have investigated the rheology of concentrated oil-in-water emulsions stabilised by silanised silica nanoparticles. The emulsions behave like highly elastic solids in response to small, uniform strains. They become unstable and begin to break down, however, on yielding. We show that the emulsion elasticity is correlated with the salt concentration in the water and hence the particle aggregation in emulsions at a given drop volume fraction. A supporting observation is that destabilisation is favoured by minimising the attractive interactions between the particles. Microscopic observations revealed that coalesced drops have anisotropic shapes and wrinkled surfaces, direct evidence of the interfacial particle layer acting like a mechanical barrier to bulk emulsion destabilisation.

Dual-network hydrogel capsules for controlled molecular transport via pH and temperature responsiveness.

We have developed innovative core-shell hydrogel capsules with a dual-network shell structure designed for precise control of molecular transport in response to external stimuli such as pH and temperature. The capsules were fabricated using a combination of microfluidic electrospray techniques and water-in-water (w/w) core-shell droplets templating. The primary network of the shell, calcium alginate (Ca-Alg), with a pKa around 3.4, exhibits sensitivity to pH. The secondary network of the shell, poly(ethylene glycol) methyl ether methacrylate (PEGMA), undergoes a volume phase transition near 60 °C. These properties enable precise molecular transport control in/out of the capsules by modulating the surface charges through varying pH and modifying pore size through temperature changes. Moreover, the dual-network shell structure not only significantly enhances the mechanical strength of the capsules but also improves their stability under external stimulus, ensuring structural integrity during the transport of molecules. This research lays the groundwork for further investigations into the multimodal stimuli-responsive hydrogel systems to control molecular transport, important in applications such as sensors and reactors for chemical cascade reactions.

The regulators of soil organic carbon mineralization upon lime and/or phosphate addition vary with depth.

Knowledge of the key factors regulating soil organic carbon (OC) mineralization in response to fertilizers and lime application is essential to understanding the effects of agricultural land management on soil OC preservation. Microbial community composition and OC availability to microorganisms have been proposed as the two most imperative factors controlling soil OC mineralization, although their relative importance is still under debate. Here we performed a laboratory incubation in combination with high-throughput sequencing and structural equation modeling to examine the mechanisms underlying the responses of OC mineralization in the topsoil and the subsoil of a volcanic soil (an Andosol) to the additions of lime and/or phosphate. Results showed that lime and/or phosphate additions induced distinct shifts in the microbial community composition and functional profiles in the topsoil and the subsoil. We found that OC mineralization relied on microbial community composition and functionality in the topsoil but was strongly related to the quality and quantity of the water-extractable OC (indicative of the OC availability) in the subsoil. These data suggest that the key regulator controlling the response of OC mineralization to lime and/or P additions shifts from microbial community composition to OC availability as soil depth increases in the Andosol. Our findings highlight the central role of mechanisms controlling soil OC mineralization in regulating the responses of mineralization to intensive agricultural management practices.

Interfacial colloidal assembly guided by optical tweezers and tuned via surface charge.

HYPOTHESIS: The size, shape and dynamics of assemblies of colloidal particles optically-trapped at an air-water interface can be tuned by controlling the optical potential, particle concentration, surface charge density and wettability of the particles and the surface tension of the solution. EXPERIMENTS: The assembly dynamics of different colloidal particle types (silica, polystyrene and carboxyl coated polystyrene particles) at an air-water interface in an optical potential were systematically explored allowing the effect of surface charge on assembly dynamics to be investigated. Additionally, the pH of the solutions were varied in order to modulate surface charge in a controllable fashion. The effect of surface tension on these assemblies was also explored by reducing the surface tension of the supporting solution by mixing ethanol with water. FINDINGS: Silica, polystyrene and carboxyl coated polystyrene particles showed distinct assembly behaviours at the air-water interface that could be rationalised taking into account changes in surface charge (which in addition to being different between the particles could be modified systematically by changing the solution pH). Additionally, this is the first report showing that wettability of the colloidal particles and the surface tension of the solution are critical in determining the resulting assembly at the solution surface.

Influence of particle concentration on multiple droplet formation in Pickering emulsions.

HYPOTHESIS: Multiphase droplets form when oil and water are mixed together at the ambivalent conditions that occur close to phase inversion. In this paper we propose a mechanism for the stabilisation of multiphase droplets by a single population of particles. EXPERIMENTS: We investigated the microstructure of emulsions formed when dodecane and water are mixed in the presence of hydrophobic fumed silica nanoparticles. We identified the range of compositions, mixing times and rates where water-in-oil-in-water emulsions are stabilised in a single mixing step. To explore how the particle availability and mixing conditions lead to multiple emulsion formation we used light scattering and microscopy techniques to probe the size and morphology of the drops, and the particle coverage of the interfaces. FINDINGS: Our key finding is that to stabilise multiphase drops there should be sufficient particles available to coat water drops that are entrained within coalescing oil droplets. The size of an entrained drop is determined by the volume of the rupturing film that forms between the oil drops. The particle coating prevents the entrained drop from escaping into the external aqueous phase. These results suggest a simple route for controlling the formation and stability of multiple emulsions for encapsulation applications.

Effect of oil soluble surfactant in emulsions stabilised by clay particles.

Although surfactants and particles are often mixed together in emulsions, the contribution of each species to the stabilisation of the oil-water interface is poorly understood. We report the results of investigations into the formation of emulsions from solutions of surfactant in oil and aqueous suspensions of laponite. Depending on the salt concentration in the aqueous suspensions, the laponite dispersed as individual disc-shaped particles, 30 nm in diameter, or flocculated into aggregates tens of micrometres in diameter. At the concentrations studied, the flocculated particles alone stabilized oil-in-water emulsions. Synergistic interactions between the particles and octadecylamine at the oil-water interface reduced the average emulsion drop size, while antagonistic interactions with octadecanoic acid enhanced coalescence processes in the emulsions. The state of particle dispersion had dramatic effects on the emulsions formed. Measurements of the oil-water interfacial tension revealed the origins of the interactions between the surfactants and particles.

Droplet Fusion in Oil-in-Water Pickering Emulsions.

We have formed compound droplets made of two or more drops of immiscible oils by temporarily destabilizing Pickering oil-in-water emulsions. The emulsions used are synergistically stabilized by mixtures of cationic surfactant and negatively-charged particles. They are highly sensitive to the concentration of surfactant present in the emulsions. We took advantage of transient droplet coalescence events that are triggered by reducing the surfactant concentration to fuse together drops of immiscible oils. This study provides guidelines for designing compound droplets by transient (or limited) coalescence in Pickering emulsions. We show that the possible geometries of particle-stabilized compound drops are determined by the interfacial tensions and relative volumes of the drops fused together. The implications of our results for designing strategies to fabricate multiphase drops are discussed.

Understanding the role of hydrogen bonding in the aggregation of fumed silica particles in triglyceride solvents.

HYPOTHESIS: Fumed silica particles are thought to thicken organic solvents into gels by aggregating to form networks. Hydrogen bonding between silanol groups on different particle surfaces causes the aggregation. The gel structure and hence flow behaviour is altered by varying the proportion of silanol groups on the particle surfaces. However, characterising the gel using rheology measurements alone is not sufficient to optimise the aggregation. We have used confocal microscopy to characterise the changes in the network microstructure caused by altering the particle surface chemistry. EXPERIMENTS: Organogels were formed by dispersing fumed silica nanoparticles in a triglyceride solvent. The particle surface chemistry was systematically varied from oleophobic to oleophilic by functionalisation with hydrocarbons. We directly visualised the particle networks using confocal scanning laser microscopy and investigated the correlations between the network structure and the shear response of the organogels. FINDINGS: Our key finding is that the sizes of the pore spaces in the networks depend on the fraction of silanol groups available to form hydrogen bonds. The reduction in the network elasticity of gels formed by methylated particles can be accounted for by the increasing pore size and tenuous nature of the networks. This is the first report that characterises the changes in the microstructure of fumed silica particle networks in non-polar solvents caused by manipulating the particle surface chemistry.

Nanoparticle adsorption and stabilisation of surfactant-free emulsions.

The formation of particle-stabilised emulsions by adding partially hydrophobised silica particles to surfactant-free oil-in-water emulsions (average drop diameter approximately 700 nm) stabilised by hydroxide ions adsorbed at the oil-water interface has been investigated. Nanoparticles (average particle diameter 18 nm) adsorbed onto the drops under alkaline conditions to produce particle-stabilised emulsions with the same drop size distribution as the surfactant-free emulsions. Unlike the surfactant-free emulsions, the particle-stabilised emulsions were stable even in acidic conditions. Strongly flocculated nanoparticles (average particle diameter 150 nm) adsorbed onto the drop surfaces under acidic conditions where the emulsions were destabilised, forming coarser particle-stabilised emulsions with micron-sized drops.

Controlling Pickering Emulsion Destabilisation: A Route to Fabricating New Materials by Phase Inversion.

The aim of this paper is to review the key findings about how particle-stabilised (or Pickering) emulsions respond to stress and break down. Over the last ten years, new insights have been gained into how particles attached to droplet (and bubble) surfaces alter the destabilisation mechanisms in emulsions. The conditions under which chemical demulsifiers displace, or detach, particles from the interface were established. Mass transfer between drops and the continuous phase was shown to disrupt the layers of particles attached to drop surfaces. The criteria for causing coalescence by applying physical stress (shear or compression) to Pickering emulsions were characterised. These findings are being used to design the structures of materials formed by breaking Pickering emulsions.

Nanostructuring Biomaterials with Specific Activities towards Digestive Enzymes for Controlled Gastrointestinal Absorption of Lipophilic Bioactive Molecules.

This review describes the development of novel lipid-based biomaterials that modulate fat digestion for the enhanced uptake of encapsulated lipophilic bioactive compounds (e.g. drugs and vitamins). Specific focus is directed towards analysing how key material characteristics affect the biological function of digestive lipases and manipulate lipolytic digestion. The mechanism of lipase action is a complex, interfacial process, whereby hydrolysis can be controlled by the ability for lipase to access and adsorb to the lipid-in-water interface. However, significant conjecture exists within the literature regarding parameters that influence the activities of digestive lipases. Important findings from recent investigations that strategically examined the interplay between the interfacial composition of the lipid microenvironment and lipolysis kinetics in simulated biophysical environments are presented. The correlation between lipolysis and the rate of solubilisation and absorption of lipophilic compounds in the gastrointestinal tract (GIT) is detailed. Greater insights into the mechanism of lipase action have provided a new approach for designing colloidal carriers that orally deliver poorly soluble compounds, directly impacting the pharmaceutical and food industries.

Destabilising Pickering emulsions by drop flocculation and adhesion.

We have investigated how emulsions of water drops coated by organoclay particles destabilise in organic solvents. The drops destabilise and the emulsions undergo a fluid-solid transition if the particles are poorly wetted by the solvent. We show that the drops adhere together and form three-dimensional networks as the fraction of the poor-quality solvent in the mixture increases. Microscopic observations revealed that the drops coalesce into buckled, non-spherical shapes in mixtures rich in poor-quality solvent. A key finding is that destabilisation is favoured under conditions where the energy of adhesion between the particle layers coating drops is comparable to the energy required to detach the particles from the drops. Rupture of the interfacial layer produces particle flocs and uncoated, unstable water drops that settle out of the emulsion.

Bioactive Hybrid Particles from Poly(D,L-lactide-co-glycolide) Nanoparticle Stabilized Lipid Droplets.

Biodegradable and bioactive hybrid particles composed of poly(D,L-lactide-co-glycolide) (PLGA) nanoparticles and medium-chain triglycerides were prepared by spray drying lipid-in-water emulsions stabilized by PLGA nanoparticles, to form PLGA-lipid hybrid (PLH) microparticles approximately 5 µm in mean diameter. The nanoparticle stabilizer was varied and mannitol was also incorporated during the preparation to investigate the effect of stabilizer charge and cryoprotectant content on the particle microstructure. An in vitro lipolysis model was used to demonstrate the particles' bioactivity by manipulating the digestion kinetics of encapsulated lipid by pancreatic lipase in simulated gastrointestinal fluid. Lipid digestion kinetics were enhanced in PLH and PLGA-lipid-mannitol hybrid (PLMH) microparticles for both stabilizers, compared to a coarse emulsion, in biorelevant media. An optimal digestion rate was observed for the negatively charged PLMH system, evidenced by a 2-fold increase in the pseudo-first-order rate constant compared to a coarse emulsion. Improved microparticle redispersion, probed by dual dye confocal fluorescence microscopy, increased the available surface area of lipid for lipase adsorption, enhancing digestion kinetics. Thereby, lipase action was controlled in hybrid microparticles by altering the surface charge and carbohydrate content. Our results demonstrate that bioactive microparticles composed of versatile and biodegradable polymeric particles and oil droplets have great potential for use in smart food and nutrient delivery, as well as safer and more efficacious oral delivery of drugs and drug combinations.

Ellipsometric Study of Monodisperse Silica Particles at an Oil-Water Interface.

Results are reported for ellipsometric measurements of hydrophobized monodisperse silica particles, with a diameter of about 25 nm, spread at the toluene-water interface. Theoretical values for the ellipsometric parameters are derived by treating the particles as a core-shell model and performing integrations of the refractive index profile through the interface using Drude's equations. With justifiable choices of the fixed parameters for the system, the agreement is good between measured and calculated values for the ellipsometric parameter Δ as a function of the amount of silica particles added to the interface. However, the results at high particle concentration at the interface are consistent either with coverage greater than a close-packed monolayer or with a monolayer with corrugations whose amplitude is less than the radius of the particles. The results show that this is not a suitable method for the determination of the contact angle of the particles at the oil-water interface.

PAA/PEO comb polymer effects on rheological properties and inter-particle forces in aqueous silica suspensions.

The effects of a poly(acrylic acid) (PAA)-poly(ethylene) (PEO) comb polymer dispersant on the rheological properties and inter-particle forces in aqueous silica suspensions have been studied under varying pH conditions. The comb polymer was found to adsorb more strongly under acidic than basic conditions, indicating that the PAA backbone of the copolymer preferentially adsorbs onto silica surfaces with the PEO "teeth" extending out from the surface into the solution. In the presence of low concentrations of copolymer, the silica suspensions were stable due to electrostatic repulsions between the silica surfaces. At higher copolymer concentrations and under neutral and basic conditions, where the copolymer interacted only weakly with silica, the suspensions showed a transition from a dispersed to weakly flocculated state and attractive forces were measured between silica surfaces. Under acidic conditions, the silica dispersion also destabilized at intermediate copolymer adsorbed density and then was re-stabilized at higher adsorbed coverage. The silica suspensions were stable at high copolymer coverage due to steric repulsions between the particles. The destabilization at intermediate coverage is thought to be due to polymer bridging between particles or possibly depletion forces.