Analysis of the presynaptic signaling mechanisms underlying the inhibition of LTP in rat dentate gyrus by the tyrosine kinase inhibitor, genistein

No abstract

Activation of adenosine A2A receptors induces TrkB translocation and increases BDNF-mediated phospho-TrkB localization in lipid rafts : implications for neuromodulation

Copyright © 2010 the authorsBrain-derived neurotrophic factor (BDNF) signaling is critical for neuronal development and transmission. Recruitment of TrkB receptors to lipid rafts has been hown to be necessary for the activation of specific signaling pathways and modulation of neurotransmitter release by BDNF. Since TrkB receptors are known to be modulated by adenosine A2A receptor activation, we hypothesized that activation of A2A receptors could influence TrkB receptor localization among different membrane microdomains. We found that adenosine A2A receptor agonists increased the levels of TrkB receptors in the lipid raft fraction of cortical membranes and potentiated BDNF-induced augmentation of phosphorylated TrkB levels in lipid rafts. Blockade of the clathrin-mediated endocytosis with monodansylcadaverine(100µM) did not modify the effects of theA2A receptor agonists but significantly impairedBDNFeffects on TrkB recruitment to lipid rafts. The effect of A2A receptor activation in TrkB localization was mimicked by 5 µM forskolin, an adenylyl cyclase activator. Also, it was blocked by the PKA inhibitors Rp-cAMPs and PKI-(14 –22), and by the Src-family kinase inhibitor PP2. Moreover, removal of endogenous adenosine or disruption of lipid rafts reduced BDNF stimulatory effects on glutamate release from cortical synaptosomes. Lipid raft integrity was also required for the effects of BDNF on hippocampal long-term potentiation at CA1 synapses. Our data demonstrate, for the first time, a BDNF-independent recruitment of TrkB receptors to lipid rafts induced by activation of adenosine A2A receptors, with functional consequences for TrkB phosphorylation and BDNF-induced modulation of neurotransmitter release and hippocampal plasticity.Supported by Fundacão para a Ciência e a Tecnologia (SFRH/BD/21374/2005 for N.A.L., SFRH/BD/21359/2005 for V.C.S., and SFRH/BPD/11528/2002 for D.B.P.) and by the European Union [European Cooperation in Science and Technology (COST) COST B30 concerted action, Neural Regeneration and Plasticity (NEREPLAS)]

SYBR Green-based Real-Time PCR targeting kinetoplast DNA can be used to discriminate between the main etiologic agents of Brazilian cutaneous and visceral leishmaniases

<p>Abstract</p> <p>Background</p> <p>Leishmaniases control has been hampered by the unavailability of rapid detection methods and the lack of suitable therapeutic and prophylactic measures. Accurate diagnosis, which can distinguish between <it>Leishmania </it>isolates, is essential for conducting appropriate prognosis, therapy and epidemiology. Molecular methods are currently being employed to detect <it>Leishmania </it>infection and categorize the parasites up to genus, complex or species level. Real-time PCR offers several advantages over traditional PCR, including faster processing time, higher sensitivity and decreased contamination risk.</p> <p>Results</p> <p>A SYBR Green real-time PCR targeting the conserved region of kinetoplast DNA minicircles was able to differentiate between <it>Leishmania </it>subgenera. A panel of reference strains representing subgenera <it>Leishmania </it>and <it>Viannia </it>was evaluated by the derivative dissociation curve analyses of the amplified fragment. Distinct values for the average melting temperature were observed, being 78.95°C ± 0.01 and 77.36°C ± 0.02 for <it>Leishmania </it>and <it>Viannia</it>, respectively (p < 0.05). Using the Neighbor-Joining method and Kimura 2-parameters, the alignment of 12 sequences from the amplified conserved minicircles segment grouped together <it>L</it>. (<it>V</it>.) <it>braziliensis </it>and <it>L</it>. (<it>V</it>.) <it>shawii </it>with a bootstrap value of 100%; while for <it>L</it>. (<it>L</it>.) <it>infantum </it>and <it>L</it>. (<it>L</it>.) <it>amazonensis</it>, two groups were formed with bootstrap values of 100% and 62%, respectively. The lower dissociation temperature observed for the subgenus <it>Viannia </it>amplicons could be due to a lower proportion of guanine/cytosine sites (43.6%) when compared to species from subgenus <it>Leishmania </it>(average of 48.4%). The method was validated with 30 clinical specimens from visceral or cutaneous leishmaniases patients living in Brazil and also with DNA samples from naturally infected <it>Lutzomyia </it>spp. captured in two Brazilian localities.</p> <p>Conclusions</p> <p>For all tested samples, a characteristic amplicon melting profile was evidenced for each <it>Leishmania </it>subgenus, corroborating the data from reference strains. Therefore, the analysis of thermal dissociation curves targeting the conserved kinetoplast DNA minicircles region is able to provide a rapid and reliable method to identify the main etiologic agents of cutaneous and visceral leishmaniases in endemic regions of Brazil.</p

In Vitro Hepatotoxic and Neurotoxic Effects of Titanium and Cerium Dioxide Nanoparticles, Arsenic and Mercury Co-Exposure

This article belongs to the Special Issue Toxicity of Nanomaterials and Legacy Contaminants: Risks to the Environment and Human Health.Considering the increasing emergence of new contaminants, such as nanomaterials, mixing with legacy contaminants, including metal(loid)s, it becomes imperative to understand the toxic profile resulting from these interactions. This work aimed at assessing and comparing the individual and combined hepatotoxic and neurotoxic potential of titanium dioxide nanoparticles (TiO2NPs 0.75-75 mg/L), cerium oxide nanoparticles (CeO2NPs 0.075-10 μg/L), arsenic (As 0.01-2.5 mg/L), and mercury (Hg 0.5-100 mg/L) on human hepatoma (HepG2) and neuroblastoma (SH-SY5Y) cells. Viability was assessed through WST-1 (24 h) and clonogenic (7 days) assays and it was affected in a dose-, time- and cell-dependent manner. Higher concentrations caused greater toxicity, while prolonged exposure caused inhibition of cell proliferation, even at low concentrations, for both cell lines. Cell cycle progression, explored by flow cytometry 24 h post-exposure, revealed that TiO2NPs, As and Hg but not CeO2NPs, changed the profiles of SH-SY5Y and HepG2 cells in a dose-dependent manner, and that the cell cycle was, overall, more affected by exposure to mixtures. Exposure to binary mixtures revealed either potentiation or antagonistic effects depending on the composition, cell type and time of exposure. These findings prove that joint toxicity of contaminants cannot be disregarded and must be further explored.This research was funded by FEDER—Fundo Europeu de Desenvolvimento Regional funds through the COMPETE 2020—Operacional Programme for Competitiveness and Internationalisation (POCI), and by Portuguese funds through FCT—Fundação para a Ciência e a Tecnologia in the framework of the project POCI-01-0145-FEDER-029651. This work was also financed by national funds through FCT—Fundação para a Ciência e a Tecnologia, I.P., in the scope of projects UIDB/04750/2020 and LA/P/0064/2020. This work was also developed within the scope of the project CICECO- Aveiro Institute of Materials, UIDB/50011/2020, UIDP/50011/2020 & LA/P/0006/2020, financed by national funds through the FCT/MEC (PIDDAC). A.T. Reis thanks the financial support of FCT through individual Grant SFRH/BPD/122112/2016. C.B. Lopes and A.C. Estrada acknowledge their research position funded by national funds (OE), through FCT—Fundação para a Ciência e a Tecnologia, I.P., in the scope of the framework contract foreseen in the numbers 4, 5 and 6 of the article 23, of the Decree-Law 57/2016, of August 29, changed by Law 57/2017, of July 19.info:eu-repo/semantics/publishedVersio

Tensile strain hardening of a metakaolin based fibre reinforced composite

Portland cement concrete is the most used building material in the world. However, its manufacture is energy-intensive and it is susceptible to harsh environments. Alternative binder systems without ordinary Portland cement, such as geopolymers or alkali-activated materials, are recently new in the Civil Engineered world. These alternative binder systems seek, among other characteristics, improved durability and environmental efficiency. The attaining of strain hardening and multiple cracking typical of Strain Hardening Cementitious Composites (SHCC) using these alternative binder systems is very attractive from a conceptual point of view, since additional endurance to certain harsh or extreme environments, as well as enhanced durability, are usually expected as two of the main outcomes. In the present work, the behaviour of two different composites was studied: an existing Engineered Cementitious Composite (ECC) and a new composite based on an alternative binder prepared with metakaolin. Polyvinyl alcohol (PVA) fibres were used in both materials. A series of experiments, including compressive and direct tensile testing were carried out to characterize and compare the mechanical properties of both materials. The results showed that the alternative binder composite, when subjected to uniaxial tension, developed multiple cracks at steadily increasing tensile stress and strain, which is also typical of ECCs showing strain hardening behaviour. The development of fibre reinforced geopolymer or alkali-activated materials showing strain hardening ability in tension may still be considered as a novel research topic, with great potential for creating new and interesting developments for Civil Engineering and structural applications, particularly the ones subjected to harsh environments

Bacterioplankton Community Shifts during a Spring Bloom of Aphanizomenon gracile and Sphaerospermopsis aphanizomenoides at a Temperate Shallow Lake

Climate change is enhancing the frequency of cyanobacterial blooms not only during summer but also in spring and autumn, leading to increased ecological impacts. The bacterioplankton community composition (BCC), in particular, is deeply affected by these blooms, although at the same time BCC can also play important roles in blooms’ dynamics. However, more information is still needed regarding BCC during species-specific cyanobacterial blooms. The goal of this study was to assess BCC succession in a hypereutrophic shallow lake (Vela Lake, Portugal) during a warm spring using a metagenomic approach to provide a glimpse of the changes these communities experience during the dominance of Aphanizomenon-like bloom-forming species. BCC shifts were studied using 16S rRNA gene metabarcoding and multivariate analyses. A total of 875 operational taxonomic units (OTUs) were retrieved from samples. In early spring, the dominant taxa belonged to Proteobacteria (mainly Alphaproteobacteria—Rickettsiales) and Bacteroidetes (Saprospirales, Flavobacteriales and Sphingobacteriales). However, at the end of May, a bloom co-dominated by cyanobacterial populations of Aphanizomenon gracile, Sphaerospermopsis aphanizomenoides and Synechococcus sp. developed and persisted until the end of spring. This led to a major BCC shift favouring the prevalence of Alphaproteobacteria (Rickettsiales and also Rhizobiales, Caulobacteriales and Rhodospirillales) and Bacteroidetes (Saprospirales, followed by Flavobacteriales and Sphingobacteriales). These results contribute to the knowledge of BCC dynamics during species-specific cyanobacterial blooms, showing that BCC is strongly affected (directly or indirectly) by Aphanizomenon-Sphaerospermopsis blooms.publishe

Emission Profiles of Carbonyl Compounds at a Brazilian Charcoal Plant

A indústria siderúrgica brasileira consome grandes quantidades de carvão vegetal. As carvoarias são responsáveis por emissões atmosféricas de compostos orgânicos voláteis tais como compostos carbonílicos (CC). Neste trabalho, foram determinados o perfil e as concentrações de CC em fase vapor durante a produção de carvão vegetal a partir de eucalipto. As amostras foram coletadas próximas aos fornos, em cartuchos de sílica revestidos com 2,4-dinitrofenilidrazina e, em seguida, extraídas e analisadas por HPLC-DAD-MS (cromatografia líquida de alta eficiência acoplada a detector de arranjo de diodo e espectrômetro de massas). Doze CC foram identificados e quantificados, entre eles acetaldeído e os pares coeluídos furfural/propanal, acroleína/propanona e 2-pentenal/pentanal. Estes três pares tiveram seus picos resolvidos por meio de espectrometria de massas para doze amostras e individualmente quantificados. Os perfis obtidos indicaram uma maior abundância do par furfural/propanal nas três fases de carbonização, seguido do acetaldeído na primeira fase e do 2-oxobutanal nas segunda e terceira fases. Furfural/propanal e acetaldeído se destacaram dentre os CC determinados nas proximidades dos fornos, com concentrações médias de 4948 e 2558 µg m -3 e concentrações máximas de 11655 e 4805 µg m -3 , respectivamente. Brazilian steel industry consumes huge amounts of charcoal. The charcoal plants are responsible for atmospheric emissions of volatile organic compounds such as carbonyl compounds (CC). In this work, the profile and the concentrations of CC in the vapor phase were determined during the charcoal production from eucalyptus. Samples were collected near kilns on silica cartridges coated with 2,4-dinitrophenylhydrazine, and then extracted and analyzed by HPLC-DAD-MS (high performance liquid chromatography coupled with diode array detector and mass spectrometry). Twelve CC were identified and quantified, amongst them acetaldehyde and the co-eluting acrolein/ propanone, furfural/propanal and 2-pentenal/pentanal pairs. These three pairs were peak resolved by mass spectra for twelve samples and thus individually quantified. The profiles obtained indicated the furfural/propanal pair as the most abundant in all three carbonization stages, followed by acetaldehyde in the first stage and 2-oxobutanal in the second and third stages. Furfural/propanal and acetaldehyde stood out among CC determined near the kilns, with average concentrations of 4948 and 2558 µg m -3 and maximum concentrations of 11655 and 4805 µg m -3 , respectively

Magnetic liposomes containing calcium ferrite nanoparticles for breast cancer therapy

In this work, calcium ferrite nanoparticles were prepared and characterized. The structural and magnetic properties of the nanoparticles were evaluated by XRD, TEM and SQUID. The synthesized nanoparticles were either entrapped in liposomes, originating aqueous magnetoliposomes (AMLs), or covered with a lipid bilayer, forming solid magnetoliposomes (SMLs). Magnetoliposomes present average diameters around 150 nm, suitable for biomedical applications. The magnetoliposomes were loaded with new potential anticancer drugs, thienopyridine derivatives, with a strong antitumor activity against breast cancer cells.This work was supported by the Portuguese Foundation for Science and Technology (FCT) in the framework of the Strategic Funding of CF-UM-UP (UID/FIS/04650/2013) and of CQUM (UID/QUI/00686/2013). FCT, POPH-QREN and FSE are acknowledged for the PhD grants of A. R. O. Rodrigues (SFRH/BD/90949/2012) and V. A. Machado (SFRH/BD/77373/2011) and for financial support to MAP-Fis Joint Doctoral Programme.info:eu-repo/semantics/publishedVersio

Differential aspects of stroke and congestive heart failure in quality of life reduction: a case series with three comparison groups

<p>Abstract</p> <p>Background</p> <p>To assess QOL of patients with stroke in comparison to other groups (caregivers and CHF patients), to identify which items of QOL are more affected on each group and what is the functional profile of patients with stroke.</p> <p>Methods</p> <p>Consecutive stroke or congestive heart failure (CHF) patients were evaluated and compared to their caregivers (caregivers). The NIH Stroke Scale (NIHSS) and EuroQoL-5D (EQ-5D) scale were applied.</p> <p>Results</p> <p>We evaluated 67 patients with stroke, 62 with CHF and 67 caregivers. For stroke patients, median NIHSS score was four. EQ-5D score was significantly worse in stroke, as compared to CHF and caregivers (0.52, 0.69 and 0.65, respectively). Mobility and usual activity domains were significantly affected in stroke and CHF patients as compared to caregivers; and self-care was more affected in stroke as compared with the other two groups.</p> <p>Conclusions</p> <p>Despite a mild neurological deficit, there was a significantly worse QOL perception in stroke as compared to CHF patients, mostly in their perception of self-care.</p

Contamination of Mesenchymal Stem-Cells with Fibroblasts Accelerates Neurodegeneration in an Experimental Model of Parkinson’s Disease

Pre-clinical studies have supported the use of mesenchymal stem cells (MSC) to treat highly prevalent neurodegenerative diseases such as Parkinson’s disease (PD) but preliminary trials have reported controversial results. In a rat model of PD induced by MPTP neurotoxin, we first observed a significant bilateral preservation of dopaminergic neurons in the substantia nigra and prevention of motor deficits typically observed in PD such as hypokinesia, catalepsy, and bradykinesia, following intracerebral administration of human umbilical cord-derived MSC (UC-MSC) early after MPTP injury. However, surprisingly, administration of fibroblasts, mesenchymal cells without stem cell properties, as a xenotransplantation control was highly detrimental, causing significant neurodegeneration and motor dysfunction independently of MPTP. This observation prompted us to further investigate the consequences of transplanting a MSC preparation contaminated with fibroblasts, a plausible circumstance in cell therapy since both cell types display similar immunophenotype and can be manipulated in vitro under the same conditions. Here we show for the first time, using the same experimental model and protocol, that transplantation of UC-MSC induced potent neuroprotection in the brain resulting in clinical benefit. However, co-transplantation of UC-MSC with fibroblasts reverted therapeutic efficacy and caused opposite damaging effects, significantly exacerbating neurodegeneration and motor deficits in MPTP-exposed rats. Besides providing a rationale for testing UC-MSC transplantation in early phases of PD aiming at delaying disease progression, our pre-clinical study suggests that fibroblasts may be common cell contaminants affecting purity of MSC preparations and clinical outcome in stem cell therapy protocols, which might also explain discrepant clinical results