Study on the Production and Re-use of Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) and Extracellular Polysaccharide by the Archaeon Haloferax mediterranei Strain DSM 1411

The halobacterium Haloferax mediterranei was used to study the production of two types of biopolymers: The biopolyester poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) was accumulated as intracellular granules, whereas an extracelluar polysaccharide was excreted in parallel to biopolyester synthesis. After production, microbial re-use and degradation of these polymers under different conditions were investigated to assess the requirements for handling the product-rich fermentation broth prior to the downstream processing for product recovery. Degradation kinetics of the polymers and the impact of different storage conditions on molar mass of PHBV were studied. It turned out that the biotechnological fermentation process can be run without any sterility precautions. No major product losses were observed without pasteurization of fermentation broth after the stop of fermentation. In addition, neither PHBV nor EPS are re-utilized by the cells for biomass formation even if the culture is maintained under conditions of carbon starvation for an extended time

Optimized electro- and wet-spinning techniques for the production of polymeric fibrous scaffolds loaded with bisphosphonate and hydroxyapatite

This research activity was aimed at the development of composite bioactive scaffolds made of biodegradable three-arm branched-star poly(ε-caprolactone) (∗PCL), hydroxyapatite nanoparticles (HNPs) and clodronate (CD), a bisphosphonate that has demonstrated efficacy in the treatment of various bone diseases and as an anti-inflammatory drug. During the experimental work, the processing conditions for the fabrication of fibrous meshes, by either electrospinning or wetspinning, were optimized. Stemming from a previous research activity on electrospinning of ∗PCL, ∗PCL/HNPs 3D meshes were developed, evaluating the influence of fabrication parameters on the fibres’ morphology. By exploiting the binding affinity of bisphosphonates for hydroxyapatite, a methodology was set up for obtaining a physical linkage between CD and HNPs, with the aim of having a dual bioactive system loaded into ∗PCL fibrous mats. Fibres loaded with either CD or CD–HNP particles were thus produced and analysed by scanning electron microscopy for their morphology and by energy dispersive X-ray spectroscopy for their elemental compositionThis work was done within the framework of the European Network of Excellence 'EXPERTISSUES', Project No. NMP3-CT-2004-500283. Professor Ramani Narayan of Michigan Biotechnology Institute and Dr Fabio Neggiani of Abiogen Pharma-Pisa are acknowledged for supplying *PCL and CD, respectively

High Performance Compostable Biocomposites Based on Bacterial Polyesters Suitable For Injection Molding and Blow Extrusion

This work deals with the design, preparation and characterization of composites based on Poly[(R)-3-hydroxybutyrate-co-(R)-3-hydroxyvalerate] and lignocellulosic filler suitable for the production of compostable and biodegradable biocomposites that mimic the thermo-mechanical and processing characteristics commonly found in those polymeric materials specially designed for injection molding and blow extrusion. The best formulation in terms of processability, thermo-mechanical properties and biodegradation behavior under mature compost conditions was the biocomposite that contained 42.8 % by wt. of Poly[(R)-3-hydroxybutyrate-co-(R)-3-hydroxyvalerate] as the major component, 5 % by wt. of lignocellulosic filler, 10 % by wt. of tributyl citrate plasticizer, 30 % by wt. of Poly(butylene adipate-co-terephthalate), 10 % by wt. of Poly(vinyl acetate), 0.2 % by wt. of Joncryl ADR-4368C a chain extender, 1 % by wt. of a primary antioxidant mix, and 1 % by wt. of nucleating agents

Biodegradable nanomats produced by electrospinning : expanding multifunctionality and potential for tissue engineering

With increasing interest in nanotechnology, development of nanofibers (n-fibers) by using the technique of electrospinning is gaining new momentum. Among important potential applications of n-fiber-based structures, scaffolds for tissue-engineering represent an advancing front. Nanoscaffolds (n-scaffolds) are closer to natural extracellular matrix (ECM) and its nanoscale fibrous structure. Although the technique of electrospinning is relatively old, various improvements have been made in the last decades to explore the spinning of submicron fibers from biodegradable polymers and to develop also multifunctional drug-releasing and bioactive scaffolds. Various factors can affect the properties of resulting nanostructures that can be classified into three main categories, namely: (1) Substrate related, (2) Apparatus related, and (3) Environment related factors. Developed n-scaffolds were tested for their cytocompatibility using different cell models and were seeded with cells for to develop tissue engineering constructs. Most importantly, studies have looked at the potential of using n-scaffolds for the development of blood vessels. There is a large area ahead for further applications and development of the field. For instance, multifunctional scaffolds that can be used as controlled delivery system do have a potential and have yet to be investigated for engineering of various tissues. So far, in vivo data on n-scaffolds are scarce, but in future reports are expected to emerge. With the convergence of the fields of nanotechnology, drug release and tissue engineering, new solutions could be found for the current limitations of tissue engineering scaffolds, which may enhance their functionality upon in vivo implantation. In this paper electrospinning process, factors affecting it, used polymers, developed n-scaffolds and their characterization are reviewed with focus on application in tissue engineering

3-iodothyronamine affects thermogenic substrates’ mobilization in brown adipocytes

We investigated the effect of 3-iodothyronamine (T1AM) on thermogenic substrates in brown adipocytes (BAs). BAs isolated from the stromal fraction of rat brown adipose tissue were exposed to an adipogenic medium containing insulin in the absence (M) or in the presence of 20 nM T1AM (M+T1AM) for 6 days. At the end of the treatment, the expression of p-PKA/PKA, p-AKT/AKT, p-AMPK/AMPK, p-CREB/CREB, p-P38/P38, type 1 and 3 beta adrenergic receptors (β1–β3AR), GLUT4, type 2 deiodinase (DIO2), and uncoupling protein 1 (UCP-1) were evaluated. The effects of cell conditioning with T1AM on fatty acid mobilization (basal and adrenergic-mediated), glucose uptake (basal and insulin-mediated), and ATP cell content were also analyzed in both cell populations. When compared to cells not exposed, M+T1AM cells showed increased p-PKA/PKA, p-AKT/AKT, p-CREB/CREB, p-P38/P38, and p-AMPK/AMPK, downregulation of DIO2 and β1AR, and upregulation of glycosylated β3AR, GLUT4, and adiponectin. At basal conditions, glycerol release was higher for M+T1AM cells than M cells, without any significant differences in basal glucose uptake. Notably, in M+T1AM cells, adrenergic agonists failed to activate PKA and lipolysis and to increase ATP level, but the glucose uptake in response to insulin exposure was more pronounced than in M cells. In conclusion, our results suggest that BAs conditioning with T1AM promote a catabolic condition promising to fight obesity and insulin resistance

Development and characterization of highly stable silver nanoparticles as novel potential antimicrobial agents for wound healing hydrogels

Recurrent microbial infections are a major cause of surgical failure and morbidity. Wound healing strategies based on hydrogels have been proposed to provide at once a barrier against pathogen microbial colonization, as well as a favorable environment for tissue repair. Nevertheless, most biocompatible hydrogel materials are more bacteriostatic than antimicrobial materials, and lack specific action against pathogens. Silver-loaded polymeric nanocomposites have efficient and selective activity against pathogenic organisms exploitable for wound healing. However, the loading of metallic nanostructures into hydrogels represents a major challenge due to the low stability of metal colloids in aqueous environments. In this context, the aim of the present study was the development of highly stable silver nanoparticles (AgNPs) as novel potential antimicrobial agents for hyaluronic acids hydrogels. Two candidate stabilizing agents obtained from natural and renewable sources, namely cellulose nanocrystals and ulvan polysaccharide, were exploited to ensure high stability of the silver colloid. Both stabilizing agents possess inherent bioactivity and biocompatibility, as well as the ability to stabilize metal nanostructures thanks to their supramolecular structures. Silver nitrate reduction through sodium borohydride in presence of the selected stabilizing agents was adopted as a model strategy to achieve AgNPs with narrow size distribution. Optimized AgNPs stabilized with the two investigated polysaccharides demonstrated high stability in phosphate buffer saline solution and strong antimicrobial activity. Loading of the developed AgNPs into photocrosslinked methacrylated hyaluronic acid hydrogels was also investigated for the first time as an effective strategy to develop novel antimicrobial wound dressing materials.This research was supported by the Portuguese Foundation for Science and Technology (FCT) under the projects PTDC/BII-BIO/28870/2017 and POCI-01-0145-FEDER-007038 (UID/Multi/ 50026/2013), and by the European Regional Development Fund (FEDER) through the “COMPETE”— Operational Program for Competitiveness factors (FCOMP-01-0124-FEDER-028120). A.R.F. thanks FCT through the Post-Doctoral scholarship SFRH/BPD/100760/2014

Exploring the Bacterial Communities of Infernaccio Waterfalls: A Phenotypic and Molecular Characterization of Acinetobacter and Pseudomonas Strains Living in a Red Epilithic Biofilm

Acquarossa river (Viterbo, Italy) was the site of a prospering Etruscan civilization thanks to metallurgical activity around 625-550 B.C. This caused the spread of heavy metals throughout the area. Rocks along the river probably act as a filter for these elements and they are covered by two different biofilms (epilithons). They differ for both color and bacterial composition. One is red and is enriched with Pseudomonas strains, while the other one is black and Acinetobacter is the most represented genus. Along the river lay the Infernaccio waterfalls, whose surrounding rocks are covered only by the red epilithon. The bacterial composition of this biofilm was analyzed through high throughput sequencing and compared to those ones of red and black epilithons of Acquarossa river. Moreover, cultivable bacteria were isolated and their phenotype (i.e., resistance against antibiotics and heavy metals) was studied. As previously observed in the case of Acquarossa river, characterization of bacterial composition of the Infernaccio red epilithon revealed that the two most represented genera were Acinetobacter and Pseudomonas. Nonetheless, these strains differed from those isolated from Acquarossa, as revealed by RAPD analysis. This work, besides increasing knowledge about the ecological properties of this site, allowed to isolate new bacterial strains, which could potentially be exploited for biotechnological applications, because of their resistance against environmental pollutants

Lie families: theory and applications

We analyze families of non-autonomous systems of first-order ordinary differential equations admitting a common time-dependent superposition rule, i.e., a time-dependent map expressing any solution of each of these systems in terms of a generic set of particular solutions of the system and some constants. We next study relations of these families, called Lie families, with the theory of Lie and quasi-Lie systems and apply our theory to provide common time-dependent superposition rules for certain Lie families.Comment: 23 pages, revised version to appear in J. Phys. A: Math. Theo

Amphiphilic pentablock copolymers and their blends with PDMS for antibiofouling coatings

Well-defined amphiphilic pentablock copolymers Siy-(EGx-FAz)2 composed of polysiloxane (Si), polyethylene glycol (EG), and perfluorohexylethyl polyacrylate (FA) blocks are synthesized by ATRP of FA monomer starting from a difunctional bromo-terminated macroinitiator. Diblock copolymers EGx-FAz are also synthesized as model systems. The block copolymers are used, either alone or blended with a PDMS matrix in varied loadings, to prepare antibiofouling coatings. Angle-resolved XPS and contact angle measurements show that the coating surface is highly enriched in fluorine content but undergoes reconstruction after contact with water. Protein adsorption experiments with human serum albumin and calf serum highlight that diblock copolymers resist protein adhesion better than do pentablock copolymers. Blending of the pentablock copolymer with PDMS results in increased protein adsorption. By contrast, the PDMS-matrix coatings show high removal percentages of sporelings of the green fouling alga Ulva linz