Microfluidic Templating of Spatially Inhomogeneous Protein Microgels

3D scaffolds in the form of hydrogels and microgels have allowed for more native cell‐culture systems to be developed relative to flat substrates. Native biological tissues are, however, usually spatially inhomogeneous and anisotropic, but regulating the spatial density of hydrogels at the microscale to mimic this inhomogeneity has been challenging to achieve. Moreover, the development of biocompatible synthesis approaches for protein‐based microgels remains challenging, and typical gelation conditions include UV light, extreme pH, extreme temperature, or organic solvents, factors which can compromise the viability of cells. This study addresses these challenges by demonstrating an approach to fabricate protein microgels with controllable radial density through microfluidic mixing and physical and enzymatic crosslinking of gelatin precursor molecules. Microgels with a higher density in their cores and microgels with a higher density in their shells are demonstrated. The microgels have robust stability at 37 °C and different dissolution rates through enzymolysis, which can be further used for gradient scaffolds for 3D cell culture, enabling controlled degradability, and the release of biomolecules. The design principles of the microgels could also be exploited to generate other soft materials for applications ranging from novel protein‐only micro reactors to soft robots.     

Modeling anisotropic light propagation in a realistic model of the human head

A Monte Carlo model capable of describing photon migration in arbitrary three-dimensional geometry with spatially varying optical properties and tissue anisotropy is presented. We use the model to explore the effects of anisotropy for optical measurements of the human head. An anisotropic diffusion equation that corresponds to our Monte Carlo model is derived, and a comparison between the Monte Carlo model and the diffusion equation solution with finite elements is given.     

Nucleated polymerisation in the presence of pre-formed seed filaments

We revisit the classical problem of nucleated polymerisation and derive a range of exact results describing polymerisation in systems intermediate between the well-known limiting cases of a reaction starting from purely soluble material and for a reaction where no new growth nuclei are formed.     

Multilocus sequence typing of Propionibacterium freudenreichii

Propionibacterium freudenreichii is used as a ripening culture in Swiss cheese manufacture. This study investigates the molecular diversity and the population structure of this bacterium via multilocus sequence typing (MLST). Internal fragments of seven genes sequenced for 113 strains of different subspecies and origins allowed the resolution of 46 sequence types (STs) with occurrence frequencies ranging from 1 to 11. The core genome of the species harbours a low level of nucleotide polymorphism. In our data, single nucleotide polymorphisms account for only 2.28% of the concatenated sequences, and the average polymorphism rate in pairwise comparisons is 0.46%. The analyses reveal quantitatively comparable contributions of recombination and mutation in nucleotide changes at core genome loci along cell lineages. Remarkably, the STs exhibit little if any dairy biotope specialization. Phenotypic characterisation of the strains, based on their aptitude to use lactose and nitrate, shows that the two previously identified subspecies (freudenreichii and shermani) do not reflect the ancestral relationships in the P. freudenreichii population. The considerable phenotypic heterogeneity, found even at the ST level, suggests instead a history of recurrent switches between phenotypes.     

The kinetics of the Cu/Cu redox couple in deep eutectic solvents

Kinetics of electron transfer of the Cu(I)/Cu(II) redox couple at a platinum electrode has been studied with chronoamperometry, cyclic voltammetry and impedance spectroscopy in a deep eutectic solvent consisting of choline chloride and ethylene glycol. At 25 °C, the reaction was found to be quasi-reversible with a relatively high rate constant k⁰ of 9.5 ± 2 × 10⁻⁴ cm s⁻¹, and a charge transfer coefficient α of 0.25 ± 0.05. Diffusion coefficients for the Cu(I) and Cu(II) complexes were determined to be 2.7 ± 0.1 × 10⁻⁷ and 1.5 ± 0.1 × 10⁻⁷ cm² s⁻¹, respectively. The viscosity of the electrolyte was 41 ± 3 mPa s. The temperature dependency was also investigated. The activation energy of mass transfer was found to be 27.7 ± 1 kJ mol⁻¹ and that of electron transfer 39 ± 7 kJ mol⁻¹. Speciation of the Cu(I) and Cu(II) complexes was determined using UV–VIS spectroscopy, and the prevailing Cu(I) complex was found to be [CuCl₃]²⁻ and that of Cu(II) [CuCl₄]²⁻.     

Hybrid knowledge-based tool for mixing equipment selection – the fuzzy neural network

The hybrid knowledge-based system proposed in this paper consists of a “stiff” segment, viz. the expert system based on the object-oriented approach, and a flexible part, viz. the neural network. Some of the input parameters of the problem and output parameters of the “stiff” system are presented as the fuzzy numbers. Detailed information is also presented about the development of the neural network. The most evident advantages of the proposed introduction of a hybrid architecture of the knowledge-based system are a faster evaluation and generation of design alternatives and support of systematic searches and storage of experience. In addition, the resulting ability to extrapolate results would be unattainable with separately acting stiff and flexible systems. A system for the estimation of the parameters of a mixing system for wastewater treatment is presented as an example to illustrate the principles of the hybrid system.     

Label-free detection of amyloid growth with microcantilever sensors

We present an approach for sensing protein aggregation using microcantilever systems. Results?from both single cantilever experiments with internal reference and multicantilever array measurements with dedicated reference cantilevers are discussed. We show that in both?cases protein aggregation on the sensor can be detected through associated changes in surface?stress.