Neurological disease in adults with Zika and chikungunya virus infection in Northeast Brazil: a prospective observational study

Background: Since 2015, the arthropod-borne viruses (arboviruses) Zika and chikungunya have spread across the Americas causing outbreaks, accompanied by increases in immune-mediated and infectious neurological disease. The spectrum of neurological manifestations linked to these viruses, and the importance of dual infection, are not known fully. We aimed to investigate whether neurological presentations differed according to the infecting arbovirus, and whether patients with dual infection had a different disease spectrum or severity. Methods: We report a prospective observational study done during epidemics of Zika and chikungunya viruses in Recife, Pernambuco, a dengue-endemic area of Brazil. We recruited adults aged 18 years or older referred to Hospital da Restauração, a secondary-level and tertiary-level hospital, with suspected acute neurological disease and a history of suspected arboviral infection. We looked for evidence of Zika, chikungunya, or dengue infection by viral RNA or specific IgM antibodies in serum or CSF. We grouped patients according to their arbovirus laboratory diagnosis and then compared demographic and clinical characteristics. Findings: Between Dec 4, 2014, and Dec 4, 2016, 1410 patients were admitted to the hospital neurology service; 201 (14%) had symptoms consistent with arbovirus infection and sufficient samples for diagnostic testing and were included in the study. The median age was 48 years (IQR 34–60), and 106 (53%) were women. 148 (74%) of 201 patients had laboratory evidence of arboviral infection. 98 (49%) of them had a single viral infection (41 [20%] had Zika, 55 [27%] had chikungunya, and two [1%] had dengue infection), whereas 50 (25%) had evidence of dual infection, mostly with Zika and chikungunya viruses (46 [23%] patients). Patients positive for arbovirus infection presented with a broad range of CNS and peripheral nervous system (PNS) disease. Chikungunya infection was more often associated with CNS disease (26 [47%] of 55 patients with chikungunya infection vs six [15%] of 41 with Zika infection; p=0·0008), especially myelitis (12 [22%] patients). Zika infection was more often associated with PNS disease (26 [63%] of 41 patients with Zika infection vs nine [16%] of 55 with chikungunya infection; p≤0·0001), particularly Guillain-Barré syndrome (25 [61%] patients). Patients with Guillain-Barré syndrome who had Zika and chikungunya dual infection had more aggressive disease, requiring intensive care support and longer hospital stays, than those with mono-infection (median 24 days [IQR 20–30] vs 17 days [10–20]; p=0·0028). Eight (17%) of 46 patients with Zika and chikungunya dual infection had a stroke or transient ischaemic attack, compared with five (6%) of 96 patients with Zika or chikungunya mono-infection (p=0·047). Interpretation: There is a wide and overlapping spectrum of neurological manifestations caused by Zika or chikungunya mono-infection and by dual infections. The possible increased risk of acute cerebrovascular disease in patients with dual infection merits further investigation. Funding: Fundação do Amparo a Ciência e Tecnologia de Pernambuco (FACEPE), EU's Horizon 2020 research and innovation programme, National Institute for Health Research. Translations: For the Portuguese and Spanish translations of the abstract see Supplementary Materials section

Smuggling tourism in the North of Portugal and Galicia

This research, which presents a qualitative methodology, has as main objective, to understand how smuggling is being remembered and portrayed today, and which strategies can be proposed by entities, to promote smuggling as one of the main tourism resources in the North of Portugal. The main findings focus on the need for the organizations to invest in this new tourism product, as well as denoting the existing constraints, concluding about the success factors for this new tourism product in the border region. To promote it, and visioning its success, the interviewees suggest the development of diversified routes along the border, because this product differentiates himself from others for it genuineness, influence the population of the border to participate more actively in the routes, to share their memories, and build new authentic experiences not previously known by tourists.info:eu-repo/semantics/publishedVersio

Effects of Gold Salt Speciation and Structure of Human and Bovine Serum Albumin on the Synthesis and Stability of Gold Nanostructures

The present study aimed to investigate the influence of albumin structure and gold speciation on the synthesis of gold nanoparticles (GNPs). The strategy of synthesis was the addition of HAuCl4 solutions at different pH values (3-12) to solutions of human and bovine serum albumins (HSA and BSA) at the same corresponding pH values. Different pH values influence the GNP synthesis due to gold speciation. Besides the inherent effect of pH on the native structure of albumins, the use N-ethylmaleimide (NEM)-treated and heat-denaturated forms of HSA and BSA provided additional insights about the influence of protein structure, net charge, and thiol group approachability on the GNP synthesis. NEM treatment, heating, and the extreme values of pH promoted loss of the native albumin structure. The formation of GNPs indicated by the appearance of surface plasmon resonance (SPR) bands became detectable from fifteen days of the synthesis processes that were carried out with native, NEM-treated and heat-denaturated forms of HSA and BSA, exclusively at pH 6 and 7. After two months of incubation, SPR band was also detected for all synthesis carried out at pH 8.0. The mean values of the hydrodynamic radius (RH) were 24 and 34 nm for GNPs synthesized with native HSA and BSA, respectively. X-ray diffraction (XRD) revealed crystallites of 13 nm. RH, XRD, and zeta potential values were consistent with GNP capping by the albumins. However, the GNPs produced with NEM-treated and heat-denaturated albumins exhibited loss of protein capping by lowering the ionic strength. This result suggests a significant contribution of non-electrostatic interactions of albumins with the GNP surface, in these conditions. The denaturation of proteins exposes hydrophobic groups to the solvent, and these groups could interact with the gold surface. In these conditions, the thiol blockage or oxidation, the latter probably favored upon heating, impaired the formation of a stable capping by thiol coordination with the gold surface. Therefore, the cysteine side chain of albumins is important for the colloidal stabilization of GNPs rather than as the reducing agent for the synthesis. Despite the presence of more reactive gold species at more acidic pH values, i.e., belo

Nanoparticle–Cell Interactions: Surface Chemistry Effects on the Cellular Uptake of Biocompatible Block Copolymer Assemblies

The development of nanovehicles for intracellular drug delivery is strongly bound to the understating and control of nanoparticles cellular uptake process, which in turn is governed by surface chemistry. In this study, we explored the synthesis, characterization, and cellular uptake of block copolymer assemblies consisting of a pH-responsive poly­[2-(diisopropyl­amino)­ethyl methacrylate] (PDPA) core stabilized by three different biocompatible hydrophilic shells (a zwitterionic type poly­(2-methacryl­oyloxyethyl phosphorylcholine) (PMPC) layer, a highly hydrated poly­(ethylene oxide) (PEO) layer with stealth effect, and an also proven nontoxic and nonimmunogenic poly­(<i>N</i>-(2-hydroxypropyl)­methacrylamide) (PHPMA) layer). All particles had a spherical core–shell structure. The largest particles with the thickest hydrophilic stabilizing shell obtained from PMPC₄₀-<i>b</i>-PDPA₇₀ were internalized to a higher level than those smaller in size and stabilized by PEO or PHPMA and produced from PEO₁₂₂-<i>b</i>-PDPA₄₃ or PHPMA₆₄-<i>b</i>-PDPA₇₂, respectively. Such a behavior was confirmed among different cell lines, with assemblies being internalized to a higher degree in cancer (HeLa) as compared to healthy (Telo-RF) cells. This fact was mainly attributed to the stronger binding of PMPC to cell membranes. Therefore, cellular uptake of nanoparticles at the sub-100 nm size range may be chiefly governed by the chemical nature of the stabilizing layer rather than particles size and/or shell thickness

Efficient Condensation of DNA into Environmentally Responsive Polyplexes Produced from Block Catiomers Carrying Amine or Diamine Groups

The intracellular delivery of nucleic acids requires a vector system as they cannot diffuse across lipid membranes. Although polymeric transfecting agents have been extensively investigated, none of the proposed gene delivery vehicles fulfill all of the requirements needed for an effective therapy, namely, the ability to bind and compact DNA into polyplexes, stability in the serum environment, endosome-disrupting capacity, efficient intracellular DNA release, and low toxicity. The challenges are mainly attributed to conflicting properties such as stability vs efficient DNA release and toxicity vs efficient endosome-disrupting capacity. Accordingly, investigations aimed at safe and efficient therapies are still essential to achieving gene therapy clinical success. Taking into account the mentioned issues, herein we have evaluated the DNA condensation ability of poly­(ethylene oxide)₁₁₃-<i>b</i>-poly­[2-(di­iso­propyl­amino)­ethyl meth­acrylate]₅₀ (PEO₁₁₃-<i>b</i>-PDPA₅₀), poly­(ethylene oxide)₁₁₃-<i>b</i>-poly­[2-(di­ethyl­amino)­ethyl meth­acrylate]₅₀ (PEO₁₁₃-<i>b</i>-PDEA₅₀), poly­[oligo­(ethylene glycol)­methyl ether meth­acrylate]₇₀-<i>b</i>-poly­[oligo­(ethylene glycol)­methyl ether meth­acrylate₁₀-<i>co</i>-2-­(diethylamino)­ethyl meth­acrylate₄₇-<i>co</i>-2-(diisopropylamino)­ethyl meth­acrylate₄₇] (POEGMA₇₀-<i>b</i>-P­(OEGMA₁₀-<i>co</i>-DEA₄₇-<i>co</i>-DPA₄₇), and poly­[oligo­(ethylene glycol)­methyl ether meth­acrylate]₇₀-<i>b</i>-poly­{oligo­(ethylene glycol)­methyl ether meth­acrylate₁₀-<i>co</i>-2-methyl­acrylic acid 2-[(2-(di­methyl­amino)­ethyl)­methyl­amino]­ethyl ester₄₄} (POEGMA₇₀-<i>b</i>-P­(OEGMA₁₀-<i>co</i>-DAMA₄₄). Block copolymers PEO₁₁₃-<i>b</i>-PDEA₅₀ and POEGMA₇₀-<i>b</i>-P­(OEGMA₁₀-<i>co</i>-DEA₄₇-<i>co</i>-DPA₄₇) were evidenced to properly condense DNA into particles with a desirable size for cellular uptake via endocytic pathways (<i>R</i>_H ≈ 65–85 nm). The structure of the polyplexes was characterized in detail by scattering techniques and atomic force microscopy. The isothermal titration calorimetric data revealed that the polymer/DNA binding is endothermic; therefore, the process in entropically driven. The combination of results supports that POEGMA₇₀-<i>b</i>-P­(OEGMA₁₀-<i>co</i>-DEA₄₇-<i>co</i>-DPA₄₇) condenses DNA more efficiently and with higher thermodynamic outputs than does PEO₁₁₃-<i>b</i>-PDEA₅₀. Finally, circular dichroism spectroscopy indicated that the conformation of DNA remained the same after complexation and that the polyplexes are very stable in the serum environment

Guillain-Barre syndrome during the Zika virus outbreak in Northeast Brazil: An observational cohort study

Objective: To determine the clinical phenotype of Guillain-Barré syndrome (GBS) after Zika virus (ZIKV) infection, the anti-glycolipid antibody signature, and the role of other circulating arthropod-borne viruses, we describe a cohort of GBS patients identified during ZIKV and chikungunya virus (CHIKV) outbreaks in Northeast Brazil. Methods: We prospectively recruited GBS patients from a regional neurology center in Northeast Brazil between December 2014 and February 2017. Serum and CSF were tested for ZIKV, CHIKV, and dengue virus (DENV), by RT-PCR and antibodies, and serum was tested for GBS-associated antibodies to glycolipids. Results: Seventy-one patients were identified. Forty-eight (68%) had laboratory evidence of a recent arbovirus infection; 25 (52%) ZIKV, 8 (17%) CHIKV, 1 (2%) DENV, and 14 (29%) ZIKV and CHIKV. Most patients with a recent arbovirus infection had motor and sensory symptoms (72%), a demyelinating electrophysiological subtype (67%) and a facial palsy (58%). Patients with a recent infection with ZIKV and CHIKV had a longer hospital admission and more frequent mechanical ventilation compared to the other patients. No specific anti-glycolipid antibody signature was identified in association with arbovirus infection, although significant antibody titres to GM1, GalC, LM1, and GalNAc-GD1a were found infrequently. Conclusion: A large proportion of cases had laboratory evidence of a recent infection with ZIKV or CHIKV, and recent infection with both viruses was found in almost one third of patients. Most patients with a recent arbovirus infection had a sensorimotor, demyelinating GBS. We did not find a specific anti-glycolipid antibody signature in association with arbovirus-related GBS