Intracellular trafficking and cellular uptake mechanism of PHBV nanoparticles for targeted delivery in epithelial cell lines

Indexación: Web of Science; Scopus; Scielo.Background: Nanotechnology is a science that involves imaging, measurement, modeling and a manipulation of matter at the nanometric scale. One application of this technology is drug delivery systems based on nanoparticles obtained from natural or synthetic sources. An example of these systems is synthetized from poly(3-hydroxybutyrate-co-3-hydroxyvalerate), which is a biodegradable, biocompatible and a low production cost polymer. The aim of this work was to investigate the uptake mechanism of PHBV nanoparticles in two different epithelial cell lines (HeLa and SKOV-3). Results: As a first step, we characterized size, shape and surface charge of nanoparticles using dynamic light scattering and transmission electron microscopy. Intracellular incorporation was evaluated through flow cytometry and fluorescence microscopy using intracellular markers. We concluded that cellular uptake mechanism is carried out in a time, concentration and energy dependent way. Our results showed that nanoparticle uptake displays a cell-specific pattern, since we have observed different colocalization in two different cell lines. In HeLa (Cervical cancer cells) this process may occur via classical endocytosis pathway and some internalization via caveolin-dependent was also observed, whereas in SKOV-3 (Ovarian cancer cells) these patterns were not observed. Rearrangement of actin filaments showed differential nanoparticle internalization patterns for HeLa and SKOV-3. Additionally, final fate of nanoparticles was also determined, showing that in both cell lines, nanoparticles ended up in lysosomes but at different times, where they are finally degraded, thereby releasing their contents. Conclusions: Our results, provide novel insight about PHBV nanoparticles internalization suggesting that for develop a proper drug delivery system is critical understand the uptake mechanism.https://jnanobiotechnology.biomedcentral.com/articles/10.1186/s12951-016-0241-

Effect of the Reduction Temperature of PdAg Nanoparticles during the Polyol Process in the Ethanol Electrooxidation Reaction

This work reports the effect of reduction temperature during the synthesis of PdAg catalysts through the polyol process and their evaluation in the ethanol electrooxidation reaction (EOR). The characterization was performed using Transmission Electron Microscopy (TEM) and X-Ray Diffraction (XRD). The electrochemical evaluation for the ethanol electrooxidation reaction was implemented in alkaline medium using chronoamperometry (CA) and cyclic voltammetry (CV). An important effect of the reduction temperature on electroactivity and catalytic stability was observed: both the maximum current density and the catalytic stability were higher in the catalyst synthesized at the highest temperature (135°C). This performance was associated with the extent of the interaction between Pd and Ag which was measured in terms of the structural expansion of Pd

Retrograde trafficking of β-dystroglycan from the plasma membrane to the nucleus

β-Dystroglycan (β-DG) is a transmembrane protein with critical roles in cell adhesion, cytoskeleton remodeling and nuclear architecture. This functional diversity is attributed to the ability of β-DG to target to, and conform specific protein assemblies at the plasma membrane (PM) and nuclear envelope (NE). Although a classical NLS and importin α/β mediated nuclear import pathway has already been described for β-DG, the intracellular trafficking route by which β-DG reaches the nucleus is unknown. In this study, we demonstrated that β-DG undergoes retrograde intracellular trafficking from the PM to the nucleus via the endosome-ER network. Furthermore, we provided evidence indicating that the translocon complex Sec61 mediates the release of β-DG from the ER membrane, making it accessible for importins and nuclear import. Finally, we show that phosphorylation of β-DG at Tyr890 is a key stimulus for β-DG nuclear translocation. Collectively our data describe the retrograde intracellular trafficking route that β-DG follows from PM to the nucleus. This dual role for a cell adhesion receptor permits the cell to functionally connect the PM with the nucleus and represents to our knowledge the first example of a cell adhesion receptor exhibiting retrograde nuclear trafficking and having dual roles in PM and NE

Impact of phenolic-rich olive leaf extract on blood pressure, plasma lipids and inflammatory markers: a randomised controlled trial

Purpose Dietary polyphenols have been demonstrated to favourably modify a number of cardiovascular risk markers such as blood pressure (BP), endothelial function and plasma lipids. We conducted a randomised, double-blind, controlled, crossover trial to investigate the effects of a phenolic-rich olive leaf extract (OLE) on BP and a number of associated vascular and metabolic measures. Methods A total of 60 pre-hypertensive [systolic blood pressure (SBP): 121–140 mmHg; diastolic blood pressure (DBP): 81–90 mmHg] males [mean age 45 (±SD 12.7 years, BMI 26.7 (±3.21) kg/m2] consumed either OLE (136 mg oleuropein; 6 mg hydroxytyrosol) or a polyphenol-free control daily for 6 weeks before switching to the alternate arm after a 4-week washout. Results Daytime [−3.95 (±SD 11.48) mmHg, p = 0.027] and 24-h SBP [−3.33 (±SD 10.81) mmHg, p = 0.045] and daytime and 24-h DBP [−3.00 (±SD 8.54) mmHg, p = 0.025; −2.42 (±SD 7.61) mmHg, p = 0.039] were all significantly lower following OLE intake, relative to the control. Reductions in plasma total cholesterol [−0.32 (±SD 0.70) mmol/L, p = 0.002], LDL cholesterol [−0.19 (±SD 0.56) mmol/L, p = 0.017] and triglycerides [−0.18 (±SD 0.48), p = 0.008] were also induced by OLE compared to control, whilst a reduction in interleukin-8 [−0.63 (±SD 1.13) pg/ml; p = 0.026] was also detected. Other markers of inflammation, vascular function and glucose metabolism were not affected. Conclusion Our data support previous research, suggesting that OLE intake engenders hypotensive and lipid-lowering effects in vivo

Seasonal drought limits tree species across the Neotropics

Within the tropics, the species richness of tree communities is strongly and positively associated with precipitation. Previous research has suggested that this macroecological pattern is driven by the negative effect of water-stress on the physiological processes of most tree species. This process implies that the range limits of taxa are defined by their ability to occur under dry conditions, and thus in terms of species distributions it predicts a nested pattern of taxa distribution from wet to dry areas. However, this ‘dry-tolerance’ hypothesis has yet to be adequately tested at large spatial and taxonomic scales. Here, using a dataset of 531 inventory plots of closed canopy forest distributed across the Western Neotropics we investigated how precipitation, evaluated both as mean annual precipitation and as the maximum climatological water deficit, influences the distribution of tropical tree species, genera and families. We find that the distributions of tree taxa are indeed nested along precipitation gradients in the western Neotropics. Taxa tolerant to seasonal drought are disproportionally widespread across the precipitation gradient, with most reaching even the wettest climates sampled; however, most taxa analysed are restricted to wet areas. Our results suggest that the ‘dry tolerance’ hypothesis has broad applicability in the world's most species-rich forests. In addition, the large number of species restricted to wetter conditions strongly indicates that an increased frequency of drought could severely threaten biodiversity in this region. Overall, this study establishes a baseline for exploring how tropical forest tree composition may change in response to current and future environmental changes in this region

Improved Performance of an Epoxy Matrix as a Result of Combining Graphene Oxide and Reduced Graphene

We present an easy and effective way to improve the mechanical properties of an epoxy matrix by reinforcing it with a combination of graphene oxide (GO) and reduced graphene oxide (RGO). These nanocomposites were prepared with different load of nanofillers: 0.1, 0.4, 0.7, 1.0 wt% and a neat epoxy. Ratios of graphene oxide and reduced graphene (GO : RGO) employed were: 0 : 1, 0.25 : 0.75, 0.5 : 0.5, 0.75 : 0.25, and 1 : 0. Results show that with only 0.4 wt% and a ratio 0.2 : 0.75 of GO : RGO, tensile strength and tensile toughness are 52% and 152% higher than neat epoxy while modulus of elasticity was improved ~20%. The obtained results suggest that it is possible achieve advantageous properties by combining graphene in oxidized and reduced conditions as it shows a synergic effect by the presence of both nanofillers

Hyperdominance in the Amazonian tree flora

The vast extent of the Amazon Basin has historically restricted the study of its tree communities to the local and regional scales. Here, we provide empirical data on the commonness, rarity, and richness of lowland tree species across the entire Amazon Basin and Guiana Shield (Amazonia), collected in 1170 tree plots in all major forest types. Extrapolations suggest that Amazonia harbors roughly 16,000 tree species, of which just 227 (1.4%) account for half of all trees. Most of these are habitat specialists and only dominant in one or two regions of the basin. We discuss some implications of the finding that a small group of species--less diverse than the North American tree flora--accounts for half of the world's most diverse tree community.This work was supported by Alberta Mennega Stichting; ALCOA Suriname; Banco de la República; Center for Agricultural Research in Suriname; Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (Plano Nacional de Pós-Graduação); Conselho Nacional de Desenvovimento Científico e Tecnológico of Brazil (CNPq) projects Programa de Pesquisas Ecológicas de Longa Duração (PELD) (558069/2009-6), Programa de Apoio a Núcleos de Excelência da Fundação de Amparo à Pesquisa do Estado do Amazonas (PRONEX-FAPEAM) (1600/2006), Áreas Úmidas, and MAUA; PELD (403792/2012-6), PPBio, CENBAM, Universal (479599/2008-4), and Universal 307807-2009-6; Fundação de Amparo À Pesquisa Do Estado Do Amazonas (APEAM) projects DCR/2006, Hidroveg with FAPESP, and PRONEX with CNPq; FAPESP; Colciencias; Duke University; Ecopetrol; FEPIM 044/2003; the Field Museum; Conservation International/DC (TEAM/INPA Manuas), Gordon and Betty Moore Foundation; Guyana Forestry Commission; Investissement d’Avenir grant of the French Agence Nationale de la Recherche (ANR) (Centre d’Étude de la Biodiversité Amazonienne ANR-10-LABX-0025); Margaret Mee Amazon Trust; Miquel fonds; National Geographic Society (7754-04, 8047-06 to P.M.J.); Netherlands Foundation for the Advancement of Tropical Research WOTRO grants WB85- 335 and W84-581; Primate Conservation Incorporated; Programme Ecosystèmes Tropicaux (French Ministry of Ecology and Sustainable Development; Shell Prospecting and Development Peru; Smithsonian Institution’s Biological Diversity of the Guiana Shield Program; Stichting het van Eeden-fonds; the Body Shop; the Ministry of the Environment of Ecuador; TROBIT; Tropenbos International; NSF (NSF-0743457 and NSF-0101775 to P.M.J.); USAID; Variety Woods Guyana; WWF-Brazil; WWF-Guianas; XIIéme Contrat de Plan Etat Région-Guyane (French Government and European Union); and grants to RAINFOR from the European Union, UK Natural Environment Research Council, the Gordon and Betty Moore Foundation, and U.S. National Geographic Society. O.L.P. is supported by a European Research Council Advanced Grant and a Royal Society Wolfson Research Merit Award

Persistent effects of pre-Columbian plant domestication on Amazonian forest composition

The extent to which pre-Columbian societies altered Amazonian landscapes is hotly debated. We performed a basin-wide analysis of pre-Columbian impacts on Amazonian forests by overlaying known archaeological sites in Amazonia with the distributions and abundances of 85 woody species domesticated by pre-Columbian peoples. Domesticated species are five times more likely to be hyperdominant than non-domesticated species. Across the basin the relative abundance and richness of domesticated species increases in forests on and around archaeological sites. In southwestern and eastern Amazonia distance to archaeological sites strongly influences the relative abundance and richness of domesticated species. Our analyses indicate that modern tree communities in Amazonia are structured to an important extent by a long history of plant domestication by Amazonian peoples

Insight into the prebiotic concept: lessons from an exploratory, double blind intervention study with inulin-type fructans in obese women

Objective To highlight the contribution of the gut microbiota to the modulation of host metabolism by dietary inulin-type fructans (ITF prebiotics) in obese women. Methods A double blind, placebo controlled, intervention study was performed with 30 obese women treated with ITF prebiotics (inulin/oligofructose 50/50 mix; n=15) or placebo (maltodextrin; n=15) for 3 months (16 g/day). Blood, faeces and urine sampling, oral glucose tolerance test, homeostasis model assessment and impedancemetry were performed before and after treatment. The gut microbial composition in faeces was analysed by phylogenetic microarray and qPCR analysis of 16S rDNA. Plasma and urine metabolic profiles were analysed by 1H-NMR spectroscopy. Results Treatment with ITF prebiotics, but not the placebo, led to an increase in Bifidobacterium and Faecalibacterium prausnitzii; both bacteria negatively correlated with serum lipopolysaccharide levels. ITF prebiotics also decreased Bacteroides intestinalis, Bacteroides vulgatus and Propionibacterium, an effect associated with a slight decrease in fat mass and with plasma lactate and phosphatidylcholine levels. No clear treatment clustering could be detected for gut microbial analysis or plasma and urine metabolomic profile analyses. However, ITF prebiotics led to subtle changes in the gut microbiota that may importantly impact on several key metabolites implicated in obesity and/or diabetes. Conclusions ITF prebiotics selectively changed the gut microbiota composition in obese women, leading to modest changes in host metabolism, as suggested by the correlation between some bacterial species and metabolic endotoxaemia or metabolomic signatures

Ribosomal protein deficiencies linked to Diamond-Blackfan anemia induce distinctive alterations of ATF4 expression

Ribosomal protein haploinsufficiency causes Diamond-Blackfan anemia (DBA) and other ribosomopathies. DBA has been linked to p53 activation and reduced GATA1 expression, but these mechanisms do not fully explain the disease. This study unveils that deficiencies in small (RPS) or large (RPL) ribosomal subunit proteins cause a p53-independent loss of ATF4, a master regulator of stress responses and erythropoiesis, by reducing the pool of actively translating ATF4 mRNAs. This defect is more pronounced in RPS deficiencies because the loss of 40S, but not 60S, subunits cause a destabilization of ATF4 transcripts. ATF4 downregulation occurs in early hematopoietic progenitors and correlates with the severity of erythroid differentiation defects in patients with DBA. It is also linked to the de-repression of fetal hemoglobin in erythroid cells, a frequent feature in patients with DBA. Our findings indicate that impaired ATF4 expression might be a primary contributor to DBA and explain the aggravated erythroid failure of RPS-mutant patients