Living bacteria rheology: population growth, aggregation patterns and cooperative behaviour under different shear flows

The activity of growing living bacteria was investigated using real-time and in situ rheology -- in stationary and oscillatory shear. Two different strains of the human pathogen Staphylococcus aureus -- strain COL and its isogenic cell wall autolysis mutant -- were considered in this work. For low bacteria density, strain COL forms small clusters, while the mutant, presenting deficient cell separation, forms irregular larger aggregates. In the early stages of growth, when subjected to a stationary shear, the viscosity of both strains increases with the population of cells. As the bacteria reach the exponential phase of growth, the viscosity of the two strains follow different and rich behaviours, with no counterpart in the optical density or in the population's colony forming units measurements. While the viscosity of strain COL keeps increasing during the exponential phase and returns close to its initial value for the late phase of growth, where the population stabilizes, the viscosity of the mutant strain decreases steeply, still in the exponential phase, remains constant for some time and increases again, reaching a constant plateau at a maximum value for the late phase of growth. These complex viscoelastic behaviours, which were observed to be shear stress dependent, are a consequence of two coupled effects: the cell density continuous increase and its changing interacting properties. The viscous and elastic moduli of strain COL, obtained with oscillatory shear, exhibit power-law behaviours whose exponent are dependent on the bacteria growth stage. The viscous and elastic moduli of the mutant have complex behaviours, emerging from the different relaxation times that are associated with the large molecules of the medium and the self-organized structures of bacteria. These behaviours reflect nevertheless the bacteria growth stage.Comment: 9 pages, 10 figure

An electro-rheological study of the nematic liquid crystal 4-n-heptyl-4'-cyanobiphenyl

An experimental and theoretical study of the electro-rheological effects observed in the nematic phase of 4-n-heptyl-4'-cyanobiphenyl has been conducted. This liquid crystal appears to be a model system, in which the observed rheological behaviour can be interpreted by the Leslie-Ericksen continuum theory for low molecular weight liquid crystals. Flow curves are illustrated at different temperatures and under the influence of an external electric field ranging from 0 to 3 kV mm-1, applied perpendicular to the direction of flow. Also presented is the apparent viscosity as a function of temperature, over similar values of electric field, obtained at different shear rates. A master flow curve has been constructed for each temperature by dividing the shear rate by the square of the electric field and multiplying by the square of a reference value of electric field. In a log-log plot, two Newtonian plateaux are found to appear at low and high shear rates, connected by a shear-thinning region. We have applied the Leslie-Ericksen continuum theory, in which the director alignment angle is a function of the electric field and the flow field boundary conditions are neglected, to determine viscoelastic parameters and the dielectric anisotropy

Design and synthesis of new inhibitors of p53–MDM2 interaction with a chalcone scaffold

The virtual screening of a library of chalcone derivatives led us to the identification of potential new MDM2 ligands. The chalcones with the best docking scores obeying the Lipinski rule of five were subsequently prepared by base-catalyzed aldol reactions. The activity of these compounds as inhibitors of p53–MDM2 interaction was investigated using a yeast-based screening assay. Using this approach two chalcones (3 and 4) were identified as putative small molecule inhibitors of p53–MDM2 interaction. The activity of both chalcones was further investigated in a panel of human tumor cells. Chalcones 3 and 4 revealed a pronounced tumor cell growth inhibitory effect on tumor cell lines. Additionally, chalcone 4 caused alterations in the cell cycle profile, induced apoptosis and increased the levels of p53, p21 and PUMA proteins in NCI-H460 cells. Computational docking studies allowed to predict that, like nutlin-3A (a well-known small-molecule inhibitor of p53–MDM2 interaction), chalcones 3 and 4 bind to the p53-binding site of MDM2. The results here presented will be valuable for the structure-based design of novel and potent p53–MDM2 inhibitors.This research was partially supported by the Strategic Funding UID/Multi/04423/2013 , ERDF , COMPETE , and FCT under the projects PTDC/MAR-BIO/4694/2014, and INNOVMAR – Innovation and Sustainability in the Management and Exploitation of Marine Resources, reference NORTE-01-0145-FEDER-000035 , Research Line NOVELMAR . This work also received financial support from the European Union (FEDER funds POCI/01/0145/FEDER/007265) and National Funds (FCT/MEC, Fundação para a Ciência e Tecnologia and Ministério da Educação e Ciência) under the Partnership Agreement PT2020 UID/QUI/50006/2013 and the FCT project PTDC/DTP-FTO/1981/2014, “PEst-C/SAU/LA0003/2013”, “NORTE-07-0162-FEDER-00018 – Contributos para o reforço da capacidade do IPATIMUP enquanto actor do sistema regional de inovação” and NORTE-07-0162-FEDER-000067 – Reforço e consolidação da capacidade infraestrutural do IPATIMUP para o sistema regional de inovação”, both supported by ON.2 – O Novo Norte, through FEDER funds under the QREN. IPATIMUP integrates the i3S Research Unit, which is partially supported by FCT. The authors also thank FCT for the grants of R.T. Lima ( SFRH/BPD/68787/2010 ), J. Soares ( SFRH/BD/78971/2011 ), and S. Gomes ( SFRH/BD/96189/2013 ; Doctoral Programme BiotechHealth), L. Raimundo ( PD/BI/113926/2015 , Doctoral Programme BiotechHealth)

Real-time rheology of actively growing bacteria

The population growth of a Staphylococcus aureus culture, an active colloidal system of spherical cells, was followed by rheological measurements, under steady-state and oscillatory shear flows. We observed a rich viscoelastic behavior as a consequence of the bacteria activity, namely, of their multiplication and density-dependent aggregation properties. In the early stages of growth (lag and exponential phases), the viscosity increases by about a factor of 20, presenting several drops and full recoveries. This allows us to evoke the existence of a percolation phenomenon. Remarkably, as the bacteria reach their late phase of development, in which the population stabilizes, the viscosity returns close to its initial value. Most probably, this is caused by a change in the bacteria physiological activity and in particular, by the decrease of their adhesion properties. The viscous and elastic moduli exhibit power-law behaviors compatible with the "soft glassy materials" model, whose exponents are dependent on the bacteria growth stage. DOI: 10.1103/PhysRevE.87.030701

A new long-term composite drug delivery system based on thermo-responsive hydrogel and nanoclay

Several problems and limitations faced in the treatment of many diseases can be overcome by using controlled drug delivery systems (DDS), where the active compound is transported to the target site, minimizing undesirable side effects. In situ-forming hydrogels that can be injected as viscous liquids and jellify under physiological conditions and biocompatible clay nanoparticles have been used in DDS development. In this work, polymer–clay composites based on Pluronics (F127 and F68) and nanoclays were developed, aiming at a biocompatible and injectable system for long-term controlled delivery of methylene blue (MB) as a model drug. MB release from the systems produced was carried out at 37◦C in a pH 7.4 medium. The Pluronic formulation selected (F127/F68 18/2 wt.%) displayed a sol/gel transition at approx. 30◦C, needing a 2.5 N force to be injected at 25◦C. The addition of 2 wt.% of Na116 clay decreased the sol/gel transition to 28◦C and significantly enhanced its viscoelastic modulus. The most suitable DDS for long-term application was the Na116-MB hybrid from which, after 15 days, only 3% of the encapsulated MB was released. The system developed in this work proved to be injectable, with a long-term drug delivery profile up to 45 days.publishersversionpublishe

Injectable hydrogels based on pluronic/water systems filled with alginate microparticles: Rheological characterization

In this paper the rheological characterization of Pluronic/water systems filled with alginate microparticles is presented. The rheological characterization of the Pluronic/water systems allowed for the choice of the best Pluronic concentration taking into account its applications as injectable hydrogels for tissue repair. The effect on the rheological behavior of the addition of alginate microparticles, to be loaded with the drug, was analyzed and the maximum concentration of microparticles determined.Spanish Ministerio de Economia y CompetitividadV Plan Propio Universidad de SevillaEuropean Union (EU)Portuguese Foundation for Science and Technology 288 UID/CTM/50025/201

A Review

This work is co-financed by FEDER, European Funds, through the COMPETE 2020 POCI and PORL, National Funds through FCT—Portuguese Foundation for Science and Technology, and POR Lisboa2020, under the projects PIDDAC (POCI-01-0145-FEDER-007688, Reference UIDB/50025/2020-2023) and PTDC/CTMCTM/30623/2017 (DREaMM). P.S. also acknowledges the individual contract CEECIND.03189.2020. C.T. acknowledges i3N for the Ph.D. grant with reference UI/BD/151541/2021. Publisher Copyright: © 2022 by the authors.In recent decades, new and improved materials have been developed with a significant interest in three-dimensional (3D) scaffolds that can cope with the diverse needs of the expanding biomedical field and promote the required biological response in multiple applications. Due to their biocompatibility, ability to encapsulate and deliver drugs, and capacity to mimic the extracellular matrix (ECM), typical hydrogels have been extensively investigated in the biomedical and biotechnological fields. The major limitations of hydrogels include poor mechanical integrity and limited cell interaction, restricting their broad applicability. To overcome these limitations, an emerging approach, aimed at the generation of hybrid materials with synergistic effects, is focused on incorporating nanoparticles (NPs) within polymeric gels to achieve nanocomposites with tailored functionality and improved properties. This review focuses on the unique contributions of clay nanoparticles, regarding the recent developments of clay-based nanocomposite hydrogels, with an emphasis on biomedical applications.publishersversionpublishe

Preliminary insights into electro-sensitive ecolubricants: A comparative analysis based on nanocelluloses and nanosilicates in castor oil

The newest generation of lubricants needs to adapt to stricter environmental policies. Simple and sustainable formulations with tunable rheological properties under the action of electric potentials may be the key. The present research explored the feasibility of producing electro-sensitive ecolubricants based on nanocellulose (crystalline and fibrillar) or nanoclay (Cloisite 15A montmorillonite and halloysite nanotubes) dispersions in castor oil, at concentrations that ranged from 2 to 6 wt.%. Broadband dielectric spectroscopy (BDS) measurements allowed for a first estimate on the electro-responsive potential of the nanofluids. The nanocelluloses and the montmorillonite suspensions presented a relaxation event in the dielectric loss, e", centered at ca. 2-4 kHz, which is related to interfacial polarization. Moreover, their actual electro-rheological (ER) effect under high electric potentials up to 4 kV/mm was assessed by determining the magnitude of the yield stress from steady flow curves at 25 °C. It was found that the nanocelluloses and the montmorillonite showed an enhancement of three orders of magnitude in their yield stress values at 4 kV/m. This enhancement was much greater than in the halloysite nanoclay, which did not exhibit any polarization). This is the starting point for the development of environmentally friendly ER lubricating fluids, based on nanocellulose and montmorillonites (layered nanosilicates), which might assist in reducing the friction and wear through the application of controlled electric fields.publishersversionpublishe