The Phylogeography of Rabies in Grenada, West Indies, and Implications for Control

In Grenada, West Indies, rabies is endemic, and is thought to be maintained in a wildlife host, the small Indian mongoose (Herpestes auropunctatus) with occasional spillover into other hosts. Therefore, the present study was undertaken to improve understanding of rabies epidemiology in Grenada and to inform rabies control policy. Mongooses were trapped island-wide between April 2011 and March 2013 and examined for the presence of Rabies virus (RABV) antigen using the direct fluorescent antibody test (dFAT) and PCR, and for serum neutralizing antibodies (SNA) using the fluorescent antibody virus neutralization test (FAVN). An additional cohort of brain samples from clinical rabies suspects submitted between April 2011 and March 2014 were also investigated for the presence of virus. Two of the 171 (1.7%) live-trapped mongooses were RABV positive by FAT and PCR, and 20 (11.7%) had SNAs. Rabies was diagnosed in 31 of the submitted animals with suspicious clinical signs: 16 mongooses, 12 dogs, 2 cats and 1 goat. Our investigation has revealed that rabies infection spread from the northeast to the southwest of Grenada within the study period. Phylogenetic analysis revealed that the viruses from Grenada formed a monophyletic clade within the cosmopolitan lineage with a common ancestor predicted to have occurred recently (6–23 years ago), and are distinct from those found in Cuba and Puerto Rico, where mongoose rabies is also endemic. These data suggest that it is likely that this specific strain of RABV was imported from European regions rather than the Americas. These data contribute essential information for any potential rabies control program in Grenada and demonstrate the importance of a sound evidence base for planning interventions

Optimization of chondroitin production in E. coli using genome scale models

Chondroitin is a natural occurring glycosaminoglycan with applications as a nutraceutical and pharmaceutical ingredient and can be extracted from animal tissues. Microbial chondroitin-like polysaccharides emerged as a safer and more sustainable alternative source. However, chondroitin titers using either natural or recombinant microorganisms are still far from meeting the increasing demand. The use of genome-scale models and computational predictions can assist the design of microbial cell factories with possible improved titers of these value-added compounds. Genome-scale models have been herein used for the first time to predict genetic modifications in Escherichia coli engineered strains that would potentially lead to improved chondroitin production. Additionally, using synthetic biology approaches, a pathway for producing chondroitin has been designed and engineered in E. coli. Afterwards, the most promising mutants identified based on bioinformatics predictions were constructed and evaluated for chondroitin production in flask fermentation. This resulted in the production of 118 mg L1 of extracellular chondroitin by overexpressing both superoxide dismutase (sodA) and a lytic murein transglycosylase (mltB). Then, batch and fed-batch fermentations at the bioreactor scale were also evaluated, in which the mutant overexpressing mltB led to an extracellular chondroitin production of 427 mg L1 and 535 mg L1, respectively. The computational approach herein described identified several potential novel targets for improved chondroitin biosynthesis, which may ultimately lead to a more efficient production of this glycosaminoglycan.info:eu-repo/semantics/publishedVersio

S100B as a potential biomarker and therapeutic target in multiple sclerosis

Multiple sclerosis (MS) pathology is characterized by neuroinflammation and demyelination. Recently, the inflammatory molecule S100B was identified in cerebrospinal fluid (CSF) and serum of MS patients. Although seen as an astrogliosis marker, lower/physiological levels of S100B are involved in oligodendrocyte differentiation/maturation. Nevertheless, increased S100B levels released upon injury may induce glial reactivity and oligodendrocyte demise, exacerbating tissue damage during an MS episode or delaying the following remyelination. Here, we aimed to unravel the functional role of S100B in the pathogenesis of MS. Elevated S100B levels were detected in the CSF of relapsing-remitting MS patients at diagnosis. Active demyelinating MS lesions showed increased expression of S100B and its receptor, the receptor for advanced glycation end products (RAGE), in the lesion area, while chronic active lesions displayed increased S100B in demyelinated areas with lower expression of RAGE in the rim. Interestingly, reactive astrocytes were identified as the predominant cellular source of S100B, whereas RAGE was expressed by activated microglia/macrophages. Using an ex vivo demyelinating model, cerebral organotypic slice cultures treated with lysophosphatidylcholine (LPC), we observed a marked elevation of S100B upon demyelination, which co-localized mostly with astrocytes. Inhibition of S100B action using a directed antibody reduced LPC-induced demyelination, prevented astrocyte reactivity and abrogated the expression of inflammatory and inflammasome-related molecules. Overall, high S100B expression in MS patient samples suggests its usefulness as a diagnostic biomarker for MS, while the beneficial outcome of its inhibition in our demyelinating model indicates S100B as an emerging therapeutic target in MS.This work was supported by Medal of Honor L’Oréal for Women in Science (FCT, UNESCO, L’Óreal) and innovation grant (Ordem dos Farmacêuticos) to AF, a post-doctoral grant from Fundação para a Ciência e Tecnologia (FCT-SFRH/BPD/96794/2013) and a DuPré Grant from the European Committee for Treatment and Research in Multiple Sclerosis (ECTRIMS) to AB, and by FCT-Pest- OE/SAU/UI4013 to iMed.ULisboa.info:eu-repo/semantics/publishedVersio

Preferência por morrer em casa e fatores associados de pessoas idosas da cidade de Belo Horizonte, Brasil

PRISMA was funded by the European Commis- sion’s Seventh Framework Programme (contract number: Health-F2-2008-201655) with the over- all aim to co-ordinate high-quality international research into end-of-life cancer care. PRISMA aimed to provide evidence and guidance on best practice to ensure that research can measure and improve outcomes for patients and families. PRISMA activities aimed to reflect the preferenc- es and cultural diversities of citizens, the clinical priorities of clinicians, and appropriately mea- sure multidimensional outcomes across settings where end–of-life care is delivered. Principal In- vestigator: Richard Harding. Scientific Director: Irene J Higginson. PRISMA members: Gwenda Albers, Barbara Antunes, Ana Barros Pinto, Clau- dia Bausewein, Dorothee Bechinger-English, Ha- mid Benalia, Emma Bennett, Lucy Bradley, Lucas Ceulemans, Barbara A Daveson, Luc Deliens, Noël Derycke, Martine de Vlieger, Let Dillen, Julia Downing, Michael Echteld, Natalie Evans, Dagny Faksvåg Haugen, Silvia Finetti, Nancy Gikaara, Barbara Gomes, Marjolein Gysels, Sue Hall, Rich- ard Harding, Irene J Higginson, Stein Kaasa, Jon- athan Koffman, Pedro Lopes Ferreira, Arantza Meñaca, Johan Menten, Natalia Monteiro Calan- zani, Fliss Murtagh, Bregje Onwuteaka-Philipsen, Roeline Pasman, Francesca Pettenati, Robert Pool, Richard A. Powell, Miel Ribbe, Katrin Sig- urdardottir, Steffen Simon, Franco Toscani, Bart Van den Eynden, Paul Vanden Berghe and Trudie van Iersel. RJ was supported by Coordination for the Improvement of Higher Education Personnel (CAPES). AF was supported by Fundação para a Ciência e a Tecnologia (FCT), within project UID/ MAT/04106/2019 (CIDMA). LS was supported by National Funds through FCT - Fundação para a Ciência e a Tecnologia within CINTESIS, R&D Unit (reference UID/IC/4255/2019).Peer reviewe

Atmospheric plasma and UV polymerisation for developing sustainable anti-adhesive polyethylene terephthalate (PET) surfaces

Enhancing the hydrophilicity of polymeric materials is an important step for achieving anti-adhesiveness. Thus, in this study, atmospheric plasma as a pre-treatment was combined with a UV grafting process to obtain a durable surface modification on polyethylene terephthalate (PET). The most promising conditions for the atmospheric plasma process were found to be 15 kW power and 4 m/min speed, leading to a contact angle reduction from 70 ± 6° to approximately 30°. However, it was observed that these values increased over time due to the ageing and washing of the PET surface, ultimately causing it to recover its initial contact angle. Therefore, the plasma-pre-treated PET samples were further modified through a UV grafting process using sodium acrylate (NaAc) and 3-sulfopropyl acrylate potassium salts (KAc). The grafted acrylate PET samples exhibited contact angles of 8 ± 3° and 28 ± 13° for NaAc and KAc, respectively, while showing durability in ageing and washing tests. The dry film thicknesses for both samples were found to be 28 ± 2 μm. Finally, the anti-adhesive properties of the NaAc- and KAc-treated surfaces were evaluated using an Escherichia coli expressing YadA, an adhesive protein from Yersinia. The modified PET surfaces were highly effective in reducing bacterial adhesion by more than 90%.This work was supported by the ViBrANT project, which received funding from the EU Horizon 2020 Research and Innovation Programme under Marie Sklowdowska-Curie (grant agreement no. 765042), and the Portuguese Foundation for Science and Technology (FCT) (grant number UIDB/04469/2020).info:eu-repo/semantics/publishedVersio

Development of a cost-effective media for biosurfactants production by Pseudomonas aeruginosa

In the last years, the textile industry has shown a growing interest in biosurfactants due to their biocompatibility , biodegradability , and versatility at various pH and temperature ranges . These compounds have found applications as softeners, wetting agents, lubricants, foam stabilizers, and even in the scouring of wool. This study aims to develop an economically efficient medium for biosurfactant production by Pseudomonas aeruginosa #112. Firstly , waste cooking oils after treatment (WCOT), a residue rich in lipids, was evaluated as an inducer of biosurfactants production . Different concentrations of these substrates (1, 2.5, 5, and 10 % w/v) were tested, and glucose was used as a carbon source. In the experiments with 1 % of WCOT it was observed a significant (p 0.05) reduction in the surface tension from 48.4 mN/m to 34.8 mN/m, suggesting the biosurfactant production . Furthermore , rice husk (RH) and vine pruning (VP) residues were identified as alternative carbon sources for biosurfactants production, when combined with WCOT . Both residues are rich in cellulose, which can be broken down into free glucose. An enzymatic pre- treatment that combines xylanase and cellulase was used to hydrolyze residues and release free glucose . The obtained results demonstrate that the combination of 1 % OUAT with hydrolyzed RH or VP resulted in a substantial (53 %) reduction in surface tension. At the end of the fermentation, 1.65 g/L and 0.26 g/L of biosurfactant were recovered for the experiments with hydrolyzed PV and RH, respectively. Additionally, the critical micelle dilution results demonstrate that the two tested media allow biosurfactant production and effective reduction of the surface tension of distilled water , even at low concentrations . This is the first report of biosurfactant production using a mixture of these three agro-industrial residues , which can be very useful for the sustainable production of these promising molecules.The authors acknowledge the financial support from integrated project be@t – Textile Bioeconomy (TCC12-i01, Sustainable Bioeconomy No. 02/C12-i01/2022), promoted by the Recovery and Resilience Plan (RRP), Next Generation EU, for the period 2021 – 2026. The authors also acknowledge the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UIDB/04469/2020 unit.info:eu-repo/semantics/publishedVersio

Modification of PET surfaces with Gum Arabic towards its bacterial anti-adhesiveness using an experimental factorial design approach

Bacterial adhesion onto hospital material surfaces still represents a big healthcare issue, being preventive measures required to mitigate this problem, such as increasing material surface hydrophilicity. In the present study, gum Arabic, a hydrophilic polysaccharide, was used to modify the surface of polyethylene terephthalate (PET). Initial water contact angle (WCA) and WCA after several washing cycles were studied as response variables by a 24 full factorial design. Several reaction parameters, such as contact time between gum Arabic and PET, gum Arabic concentration, curing temperature and curing time for PET modification were investigated. The most significant parameters were found to be the curing temperature and curing time. The optimized parameters led to a WCA reduction from 70° to 27°. The modified PET samples were characterized using several techniques including AFM, colorimetric, ATR-FTIR and contact angle which further confirmed a successful surface modification. Furthermore, bacterial adhesion assays have clearly shown that the treated PET material was highly effective in preventing the bacterial adhesion of Escherichia coli expressing YadA, an adhesive protein from Yersinia so-called Yersinia adhesin. The use of design of experiments techniques allowed for successfully attaining a PET material with a high bacterial anti-adhesiveness, using a simple grafting approach.This work was supported by the ViBrANT project that received funding from the EU Horizon 2020 Research and Innovation Programme under the Marie Sklowdowska-Curie, Grant agreement no 765042 and the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UIDB/04469/2020.info:eu-repo/semantics/publishedVersio

Species-Specific Codon Context Rules Unveil Non-Neutrality Effects of Synonymous Mutations

Codon pair usage (codon context) is a species specific gene primary structure feature whose evolutionary and functional roles are poorly understood. The data available show that codon-context has direct impact on both translation accuracy and efficiency, but one does not yet understand how it affects these two translation variables or whether context biases shape gene evolution.Here we study codon-context biases using a set of 72 orthologous highly conserved genes from bacteria, archaea, fungi and high eukaryotes to identify 7 distinct groups of codon context rules. We show that synonymous mutations, i.e., neutral mutations that occur in synonymous codons of codon-pairs, are selected to maintain context biases and that non-synonymous mutations, i.e., non-neutral mutations that alter protein amino acid sequences, are also under selective pressure to preserve codon-context biases.Since in vivo studies provide evidence for a role of codon context on decoding fidelity in E. coli and for decoding efficiency in mammalian cells, our data support the hypothesis that, like codon usage, codon context modulates the evolution of gene primary structure and fine tunes the structure of open reading frames for high genome translational fidelity and efficiency in the 3 domains of life

Can superhydrophobic PET surfaces prevent bacterial adhesion?

Prevention of bacterial adhesion is a way to reduce and/or avoid biofilm formation, thus restraining its associated infections. The development of repellent anti-adhesive surfaces, such as superhydrophobic surfaces, can be a strategy to avoid bacterial adhesion. In this study, a polyethylene terephthalate (PET) film was modified by in situ growth of silica nanoparticles (NPs) to create a rough surface. The surface was further modified with fluorinated carbon chains to increase its hydrophobicity. The modified PET surfaces presented a pronounced superhydrophobic character, showing a water contact angle of 156° and a roughness of 104 nm (a considerable increase comparing with the 69° and 4.8 nm obtained for the untreated PET). Scanning Electron Microscopy was used to evaluate the modified surfaces morphology, further confirming its successful modification with nanoparticles. Additionally, a bacterial adhesion assay using an Escherichia coli expressing YadA, an adhesive protein from Yersinia so-called Yersinia adhesin A, was used to assess the anti-adhesive potential of the modified PET. Contrarily to what was expected, adhesion of E. coli YadA was found to increase on the modified PET surfaces, exhibiting a clear preference for the crevices. This study highlights the role of material micro topography as an important attribute when considering bacterial adhesion.This work was supported by the ViBrANT project that received funding from the EU Horizon 2020 Research and Innovation Programme under the Marie Sklowdowska-Curie, Grant agreement no 765042 and the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UIDB/04469/2020.info:eu-repo/semantics/publishedVersio

Collagen-coated magnetic nanoparticles to capture pathogens from biological samples

Conventional methods of diagnosing bacterial infections such as microbial culture and molecular techniques, while highly sensitive, rely on expensive equipment and highly skilled operators. There is a need for affordable and portable diagnostic systems which are simple to operate while preserving reliability. Pre-enrichment of bacteria present in a sample coupled with subsequent bacterial identification steps can serve as a simple yet effective diagnostic technique. Magnetic nanoparticles (MNPs) coated with collagen were used to demonstrate enrichment of E.coli recombinantly expressing adhesins YadA and UspA2 since these adhesins are known to target and adhere to host collagen. The MNPs were synthesized chemically and characterized by Fourier transform infrared spectroscopy and Dynamic light scattering and the most stable MNPs were selected. Adhesion assays were performed together with fluorescent microscopy imaging to assess the pre-enrichment of bacteria by the collagen MNPs. Capture of bacteria by the collagen MNPs was successfully observed and capture efficiency of the collagen MNPs for E.coli YadA and E.coli UspA2 was calculated to be 50% and 68% respectively.VibrANT H2020-MSCA-ITN-2017, agreement no. 765042. FCT UIDB/04469/2020 unit and BioTecNorte operation (NORTE-01-0145-FEDER-000004).info:eu-repo/semantics/publishedVersio