A continuous solvent- and oil-free method to prepare injectable PEGylated fibrinogen cell-laden microparticles

Injectable biomaterials with in situ cross-linking reactions have been suggested to minimize the invasiveness associated with most implantation procedures. However, problems related with the rapid liquid-to-gel transition reaction can arise because it is difficult to predict the reliability of the reaction and its end products, as well as to mitigate cytotoxicity to the surrounding tissues. An alternative minimally invasive approach to deliver solid implants in vivo is based on injectable microparticles, which can be processed in vitro with high fidelity and reliability, while showing low cytotoxicity. Their delivery to the defect can be performed by injection through a small diameter syringe needle. We present a new methodology for the continuous, solvent- and oil-free production of photopolymerizable microparticles containing encapsulated human dermal fibroblasts. A precursor solution of cells in photo-reactive PEG-fibrinogen (PF) polymer was transported through a transparent injector exposed to light-irradiation before being atomized in a jet-in-air nozzle. Shear rheometry data provided the cross-linking kinetics of each PF/cell solution, which was then used to determine the amount of irradiation required to partially polymerize the mixture prior to atomization. The partially polymerized drops fell into a gelation bath for further polymerization. The system was capable of producing cell-laden microparticles with high cellular viability, with an average diameter of between 88.1 µm to 347.1 µm and a dispersity of between 1.1 and 2.4, depending on the parameters chosen.

Tendon regeneration through a scaffold-free approach: development of tenogenic magnetic hASCs sheets

Tendon's regeneration is limited, demanding for cell-based strategies to fully restore their functionality upon injury. The concept of magnetic force-based TE(1), generally using magnetic nanoparticles may enable, for example, stem cell stimulation and/or remote control over TE constructs. Thus, we originally propose the development of magnetic cell sheets (magCSs) with tenogenic capability, aimed at promoting tendon's regeneration. A Tenomodulin (TNMD+) subpopulation was sorted from human adipose stem cells (hASCs), using TNMD-coated immunomagnetic beads(2) and used as cell source for the development of magCSs. Briefly, cells were labeled with iron oxide composite particles (Micromod) and cultured for 7 days in α-MEM medium with or without magnetic stimulation provided by a magnetic device (nanoTherics). CSs were retrieved from the plates using magnet attraction as contiguous sheets of cells within its own deposited ECM.

Engineering of fatty acid production and secretion in Saccharomyces cerevisiae

Tese de Doutoramento em Ciências - Especialidade em Biologia

Connecting free volume with shape memory properties in noncytotoxic gamma-irradiated polycyclooctene

The free volume holes of a shape memory polymer have been analysed considering that the empty space between molecules is necessary for the molecular motion, and the shape memory response is based on polymer segments acting as molecular switches through variable flexibility with temperature or other stimuli. Therefore, thermomechanical analysis (TMA) and positron annihilation lifetime spectroscopy (PALS) have been applied to analyse shape recovery and free volume hole sizes in gamma irradiated polycyclooctene (PCO) samples, as a non-cytotoxic alternative to more conventional PCO crosslinked via peroxide for future applications in medicine. Thus, a first approach relating structure, free volume holes and shape memory properties in gamma irradiated PCO is presented. The results suggest that free volume holes caused by gamma irradiation in PCO samples facilitate the recovery process by improving movement of polymer chains and open t possibilities for the design and control of the macroscopic response.

Filamentous fungi in free water and biofilms from Brazilian drinking water systems

In some regions of Brazil, especially where the water is scarce, drinking water is stored in water storage tanks. This practice gives the consumer the guarantee of available water. The water storage conditions such as the exposure to hot weather when the tanks are on rooftops allow the development of microorganisms and microbial biofilms which can deteriorate the water quality and increase the risk to human health [1,2]. This study describes the filamentous fungi (FF) detected in free water and biofilms in drinking water storage tanks in Recife - Pernambuco, Brazil. Five sampling times in triplicate were performed at two distinct points. Colony-forming units (CFU) of FF fungi were determined with 0.45 µm filtration membranes using peptone glucose rose Bengal agar (PGRBA). From the 30 samples analysed a total of 1136 CFU were obtained. The water biofilms were collected from samplers consisting of polyethylene coupons, previously installed in the reservoirs. These coupons were transferred to PGRBA plates and incubated using with the same conditions described for free FF. For the in situ detection of FF in biofilms the Calcofluor White staining technique was used. This procedure demonstrated FF forming biofilms on the surfaces of the coupons. Brazilian legislation does not define limits for FF in drinking water. However considering the potential risk of fungal contamination, the data obtained in this study will contribute to developing future quantitative and qualitative parameters for the presence of fungi in drinking water distribution systems in Brazil. [1] HageskaL, G, Lima, N, Skaar, I. The study of fungi in drinking water. Mycological Research, 113, 2009, 165-172. [2] Skaar I, Hageskal G. Fungi in Drinking Water. In.: Paterson RRM, Lima N. (Eds.) Molecular Biology of Food and Water Borne Mycotoxigenic and Mycotic Fungi. CRC Press, Taylor & Francis Group, Boca Raton, 2015, 597-606.

Rational manipulation of mRNA folding free energy allows rheostat control of pneumolysin production by Streptococcus pneumoniae

Rational manipulation of mRNA folding free energy allows rheostat control of pneumolysin production by Streptococcus pneumoniae

Free-standing multilayered membranes based on graphene and natural polymers for biomedical applications

Dissertação de mestrado integrado em Engenharia Biomédica (área de especialização em Biomateriais, Reabilitação e Biomecânica)

Exploring the properties of genetically engineered silk-elastin-like protein films

Free standing films of a genetically engineered silk-elastin-like protein (SELP) were prepared using water and formic acid as solvents. Exposure to methanol-saturated air promoted the formation of aggregated β-strands rendering aqueous insolubility and improved the mechanical properties leading to a 10-fold increase in strain-to-failure. The films were optically clear with resistivity values similar to natural rubber and thermally stable up to 180 °C. Addition of glycerol showed to enhance the flexibility of SELP/glycerol films by interacting with SELP molecules through hydrogen bonding, interpenetrating between the polymer chains and granting more conformational freedom. This detailed characterization provides cues for future and unique applications using SELP based biopolymers.

Oxygen mass transfer impact on citric acid production by Yarrowia lipolytica from crude glycerol

Production of citric acid from crude glycerol from biodiesel industry, in batch cultures of Yarrowia lipolytica W29 was performed in a lab-scale stirred tank bioreactor in order to assess the effect of oxygen mass transfer rate in this bioprocess. An empirical correlation was proposed to describe oxygen volumetric mass transfer coefficient (kLa) as a function of operating conditions (stirring speed and specific air flow rate) and cellular density. kLa increased according with a power function with specific power input and superficial gas velocity, and slightly decreased with cellular density. The increase of initial kLa from 7 h-1 to 55 h-1 led to 7.8-fold increase of citric acid final concentration. Experiments were also performed at controlled dissolved oxygen (DO) and citric acid concentration increased with DO up to 60% of saturation. Thus, due to the simpler operation setting an optimal kLa than at controlled DO, it can be concluded that kLa is an adequate parameter for the optimization of citric acid production from crude glycerol by Y. lipolytica and to be considered in bioprocess scale-up. Our empirical correlation, considering the operating conditions and cellular density, will be a valid tool for this purpose.

Modelling and verifying smell-free architectures with the Archery language

Architectural (bad) smells are design decisions found in software architectures that degrade the ability of systems to evolve. This paper presents an approach to verify that a software architecture is smellfree using the Archery architectural description language. The language provides a core for modelling software architectures and an extension for specifying constraints. The approach consists in precisely specifying architectural smells as constraints, and then verifying that software architectures do not satisfy any of them. The constraint language is based on a propositional modal logic with recursion that includes: a converse operator for relations among architectural concepts, graded modalities for describing the cardinality in such relations, and nominals referencing architectural elements. Four architectural smells illustrate the approach.

Filamentous fungi occurrence in free water and biofilms from drinking water storage tanks

In some regions of Brazil, especially where the water is scarce, drinking water is stored in water storage tanks. This practice gives the consumer the guarantee of available water. The water storage conditions such as the exposure to hot weather when the tanks are on rooftops allow the development of microorganisms and microbial biofilms which can deteriorate the water quality and increase the risk to human health [1,2]. This study describes the filamentous fungi (FF) detected in free water and biofilms in drinking water storage tanks in Recife - Pernambuco, Brazil. Five sampling times in triplicate were performed at two distinct points. Colony-forming units (CFU) of FF fungi were determined with 0.45 μm filtration membranes using peptone glucose rose Bengal agar (PGRBA). From the 30 samples analysed a total of 1136 CFU were obtained. The water biofilms were collected from samplers consisting of polyethylene coupons, previously installed in the reservoirs. These coupons were transferred to PGRBA plates and incubated using with the same conditions described for free FF. For the in situ detection of FF in biofilms the Calcofluor White staining technique was used. This procedure demonstrated FF forming biofilms on the surfaces of the coupons. Brazilian legislation does not define limits for FF in drinking water. However considering the potential risk of fungal contamination, the data obtained in this study will contribute to developing future quantitative and qualitative parameters for the presence of fungi in drinking water distribution systems in Brazil.

Citric acid production by Y. lipolytica W29 from crude glycerol: strain improvement by mutagenesis

[Excerpt] Citric acid, an important and versatile organic acid extensively used in several industries, is originally produced by Aspergillus niger in submerged fermentation from molasses [1]. However, Yarrowia lipolytica have been studied and demonstrate a great potential as citric acid producer from several carbon sources [1–5] including crude glycerol, a low cost byproduct from the biodiesel industry [6]. The simultaneous production of the isomer isocitric acid is the major problem in using this yeast in the citric acid production. (...)

Contributos do postal ilustrado para a história do Free Card

Dissertação de mestrado em Ciências da Comunicação (área de especialização em Publicidade e Relações Públicas)

Valorization of eucalyptus wood by glycerol-organosolv pretreatment within the biorefinery concept: an integrated and intensified approach

The efficient utilization of lignocellulosic biomass and the reduction of production cost are mandatory to attain a cost-effective lignocellulose-to-ethanol process. The selection of suitable pretreatment that allows an effective fractionation of biomass and the use of pretreated material at high-solid loadings on saccharification and fermentation (SSF) processes are considered promising strategies for that purpose. Eucalyptus globulus wood was fractionated by organosolv process at 200 C for 69 min using 56% of glycerol-water. A 99% of cellulose remained in pretreated biomass and 65% of lignin was solubilized. Precipitated lignin was characterized for chemical composition and thermal behavior, showing similar features to commercial lignin. In order to produce lignocellulosic ethanol at high-gravity, a full factory design was carried to assess the liquid to solid ratio (3e9 g/g) and enzyme to solid ratio (8e16 FPU/g) on SSF of delignified Eucalyptus. High ethanol concentration (94 g/L) corresponding to 77% of conversion at 16FPU/g and LSR ¼ 3 g/g using an industrial and thermotolerant Saccharomyces cerevisiae strain was successfully produced from pretreated biomass. Process integration of a suitable pretreatment, which allows for whole biomass valorization, with intensified saccharification-fermentation stages was shown to be feasible strategy for the co-production of high ethanol titers, oligosaccharides and lignin paving the way for cost-effective Eucalyptus biorefinery.

Sensitive label-free electron chemical capacitive transducer signal for D-dimer electroanalyses

Supplementary data associated with this article can be found, in the online version, at: http://dx.doi.org/10.1016/j.electacta.2015.09.169.