Chondrogenic differentiation within magnetic-responsive multilayered liquified capsules containing collagen II/TFG-β3 coated microparticles

The use of stem cells is a promising therapeutic approach for the substantial challenge to regenerate cartilage. Considering the two prerequisites, namely the use of a 3D system to enable the chondrogenic differentiation and growth factors to avoid dedifferentiation, the diffusion efficiency of essential biomolecules is an intrinsic issue. We already proposed a liquified bioencapsulation system containing solid microparticles as cell adhesion sites1. Here, we intend to use the optimized system towards chondrogenic differentiation by encapsulating stem cells and collagenII-TGF-β3 PLLA microparticles. As a proof-of-concept, magnetite-nanoparticles were incorporated into the multilayered membrane. This can be a great advantage after implantation procedures to fixate the capsules in situ with the held of an external magnetic patch and for the follow-up through imaging. Results showed that the production of glycosaminoglycans and the expression of cartilage-relevant markers (collagen II, Sox9, aggrecan, and COMP) increased up to 28 days, while hypertrophic (collagen X) and fibrotic (collagen I) markers were downregulated. The presence of nanofibers in the newly deposited ECM was visualized by SEM, which resembles the collagen fibrils of native cartilage. The presence of the major constituent of cartilage, collagen II, was detected by immunocytochemistry and afranin-O and alcian blue stainings revealed a basophilic ECM deposition, which is characteristic of neocartilage. These findings suggest that the proposed system may provide a suitable environment for chondrogenic differentiation.

Comparison of different numerical methods for the solution of the time-fractional reaction-diffusion equation with variable diffusion coefficient

In this work we perform a comparison of two different numerical schemes for the solution of the time-fractional diffusion equation with variable diffusion coefficient and a nonlinear source term. The two methods are the implicit numerical scheme presented in [M.L. Morgado, M. Rebelo, Numerical approximation of distributed order reaction- diffusion equations, Journal of Computational and Applied Mathematics 275 (2015) 216-227] that is adapted to our type of equation, and a colocation method where Chebyshev polynomials are used to reduce the fractional differential equation to a system of ordinary differential equations

Designing cnc knit for hybrid membrane and bending active structures

Recent advances in computation allow for the integration of design and simulation of highly interrelated systems, such as hybrids of structural membranes and bending active elements. The engaged complexities of forces and logistics can be mediated through the development of materials with project specific properties and detailing. CNC knitting with high tenacity yarn enables this practice and offers an alternative to current woven membranes. The design and fabrication of an 8m high fabric tower through an interdisciplinary team of architects, structural and textile engineers, allowed to investigate means to design, specify, make and test CNC knit as material for hybrid structures in architectural scale. This paper shares the developed process, identifies challenges, potentials and future work.

Peptaibolin analogues by incorporation of α,α-dialkylglycines: synthesis and study of their membrane permeating ability

Analogues of Peptaibolin, a peptaibol with antibiotic activity, incorporating α,α-dialkylglycines (Deg, Dpg, and Ac6c) at selected positions were synthesised by MW-SPPS and fully characterized. A control analogue incorporating L-alanine was also prepared. The native peptide and the analogues were studied by fluorescence spectroscopy for their membrane permeating activity. Small unilamellar vesicles (SUVs) of egg phosphatidylcholine/ cholesterol (70:30) containing an encapsulated fluorescence probe (6-carboxyfluorescein) were used as membrane models. The assays of carboxyfluorescein release from SUVs upon peptide addition showed that Peptaibolin-Dpg and Peptaibolin-Ac6c are the most active peptides. These results indicate that the structure of the α,α-dialkylglycines is crucial for the membrane permeating ability of these Peptaibolin analogues.

Study of grapevine solute transporters involved in berry quality. A biochemical and molecular approach

Tese de Doutoramento em Biologia de Plantas

Separation by thermal diffusion in a rotary column

A rotary thermal diffusion column with the inner cylinder rotating and the outer cylinder static was used to separate n-heptane-benzene mixtures at different speeds of rotation. The results show that the column efficiency depends on the speed of rotation. For the optimum speed the increase in efficiency relative to the static column was of the order of 8%. The role of the geometric irregularities in the annulus width on performance of the rotary column is also discussed.

Nonsymmetrical separations in thermal diffusion columns

The influence of the feed composition upon the actual degrees of separation attained at the top and bottom sections of a thermogravitational column is discussed using the classical phenomenological theory of Furry, Jones, and Onsager. It is shown that, except for a feed composition of C 0 = 0.5 (mass fraction), the separation profile is nonsymmetric, i.e., the separations in the top and bottom sections of the column are nonsymmetric with respect to the feed composition, the asymmetry increasing as the feed composition moves away from C 0 = 0.5. An equation for the determination of the optimum feed location as a function of the feed composition is derived.

Some observations on asymmetrical separation in thermal diffusion columns

Data have been obtained in steady-state batch operated thermogravitational separation columns using different binary mixtures to test the theory recently published by Morgado et al. The experimental results confirm that separations by thermal diffusion are asymmetrical except when the initial concentration is 0.5 and that the asymmetry is larger as the initial concentration deviates from 0.5 and as the separation potential increases.

The application of the equivalent annulus width concept in thermal diffusion separations

The equivalent annulus width concept is used to characterize a small commercial thermogravitational hermal diffusion column and its validity checked experimentally by separating batchwise in the column mixtures of n-heptanebenzene with different initial concentrations. The equation of Ruppell and Coull was used to analyse the data in the short separation times range and determine the equivalent annulus width. Good agreement was obtained between the experimental and predicted time-separation curves when using the equivalent annulus width value and on averaged value of the thermal diffusion constant. A new method is presented for the simultaneous determination of the equivalent annulus width and the thermal diffusion constant of a binary mixture from a single set of experimental data.

Single molecule simulation of diffusion and enzyme kinetics

This work presents a molecular-scale agent-based model for the simulation of enzymatic reactions at experimentally measured concentrations. The model incorporates stochasticity and spatial dependence, using diffusing and reacting particles with physical dimensions. We developed strategies to adjust and validate the enzymatic rates and diffusion coefficients to the information required by the computational agents, i.e., collision efficiency, interaction logic between agents, the time scale associated with interactions (e.g., kinetics), and agent velocity. Also, we tested the impact of molecular location (a source of biological noise) in the speed at which the reactions take place. Simulations were conducted for experimental data on the 2-hydroxymuconate tautomerase (EC 5.3.2.6, UniProt ID Q01468) and the Steroid Delta-isomerase (EC 5.3.3.1, UniProt ID P07445). Obtained results demonstrate that our approach is in accordance to existing experimental data and long-term biophysical and biochemical assumptions.