Paracrine effect of carbon monoxide - astrocytes promote neuroprotection through purinergic signaling in mice

© 2016. Published by The Company of Biologists Ltd.The neuroprotective role of carbon monoxide (CO) has been studied in a cell-autonomous mode. Herein, a new concept is disclosed - CO affects astrocyte-neuron communication in a paracrine manner to promote neuroprotection. Neuronal survival was assessed when co-cultured with astrocytes that had been pre-treated or not with CO. The CO-pre-treated astrocytes reduced neuronal cell death, and the cellular mechanisms were investigated, focusing on purinergic signaling. CO modulates astrocytic metabolism and extracellular ATP content in the co-culture medium. Moreover, several antagonists of P1 adenosine and P2 ATP receptors partially reverted CO-induced neuroprotection through astrocytes. Likewise, knocking down expression of the neuronal P1 adenosine receptor A2A-R (encoded by Adora2a) reverted the neuroprotective effects of CO-exposed astrocytes. The neuroprotection of CO-treated astrocytes also decreased following prevention of ATP or adenosine release from astrocytic cells and inhibition of extracellular ATP metabolism into adenosine. Finally, the neuronal downstream event involves TrkB (also known as NTRK2) receptors and BDNF. Pharmacological and genetic inhibition of TrkB receptors reverts neuroprotection triggered by CO-treated astrocytes. Furthermore, the neuronal ratio of BDNF to pro-BDNF increased in the presence of CO-treated astrocytes and decreased whenever A2A-R expression was silenced. In summary, CO prevents neuronal cell death in a paracrine manner by targeting astrocytic metabolism through purinergic signaling.publishersversionpublishe

Physicochemical properties and cell viability of shrimp chitosan films as affected by film casting solvents. I-potential use as wound dressing

: Chitosan solubility in aqueous organic acids has been widely investigated. However, most of the previous works have been done with plasticized chitosan films and using acetic acid as the film casting solvent. In addition, the properties of these films varied among studies, since they are influenced by different factors such as the chitin source used to produce chitosan, the processing variables involved in the conversion of chitin into chitosan, chitosan properties, types of acids used to dissolve chitosan, types and amounts of plasticizers and the film preparation method. Therefore, this work aimed to prepare chitosan films by the solvent casting method, using chitosan derived from Litopenaeus vannamei shrimp shell waste, and five different organic acids (acetic, lactic, maleic, tartaric, and citric acids) without plasticizer, in order to evaluate the effect of organic acid type and chitosan source on physicochemical properties, degradation and cytotoxicity of these chitosan films. The goal was to select the best suited casting solvent to develop wound dressing from shrimp chitosan films. Shrimp chitosan films were analyzed in terms of their qualitative assessment, thickness, water vapor permeability (WVP), water vapor transmission rate (WVTR), wettability, tensile properties, degradation in phosphate buffered saline (PBS) and cytotoxicity towards human fibroblasts using the resazurin reduction method. Regardless of the acid type employed in film preparation, all films were transparent and slightly yellowish, presented homogeneous surfaces, and the thickness was compatible with the epidermis thickness. However, only the ones prepared with maleic acid presented adequate characteristics of WVP, WVTR, wettability, degradability, cytotoxicity and good tensile properties for future application as a wound dressing material. The findings of this study contributed not only to select the best suited casting solvent to develop chitosan films for wound dressing but also to normalize a solubilization protocol for chitosan, derived from Litopenaeus vannamei shrimp shell waste, which can be used in the pharmaceutical industry.info:eu-repo/semantics/publishedVersio

Molecular clouds in M51 from high-resolution extinction mapping

Here we present the cloud population extracted from M51, following the application of our new high-resolution dust extinction technique to the galaxy (Faustino Vieira et al. 2023). With this technique, we are able to image the gas content of the entire disc of M51 down to 5 pc (0.14"), which allows us to perform a statistical characterisation of well-resolved molecular cloud properties across different large-scale dynamical environments and with galactocentric distance. We find that cloud growth is promoted in regions in the galaxy where shear is minimised; i.e. clouds can grow into higher masses (and surface densities) inside the spiral arms and molecular ring. We do not detect any enhancement of high-mass star formation towards regions favourable to cloud growth, indicating that massive and/or dense clouds are not the sole ingredient for high-mass star formation. We find that in the spiral arms there is a significant decline of cloud surface densities with increasing galactocentric radius, whilst in the inter-arm regions they remain relatively constant. We also find that the surface density distribution for spiral arm clouds has two distinct behaviours in the inner and outer galaxy, with average cloud surface densities at larger galactocentric radii becoming similar to inter-arm clouds. We propose that the tidal interaction between M51 and its companion (NGC 5195) - which heavily affects the nature of the spiral structure - might be the main factor behind this.Comment: MNRAS accepted, 20 pages, 16 figure

A high-resolution extinction mapping technique for face-on disc galaxies

We present a new dust extinction technique with which we are able to retrieve parsec-scale gas surface density maps for entire nearby galaxies. The method measures the dust attenuation in optical bands on a pixel-by-pixel basis against a smoothed, reconstructed stellar distribution. The contribution of foreground light along the line-of-sight is calibrated using dust emission observations, assuming that the dust sits in a layer close to the mid-plane of the face-on galaxy. Here, we apply this technique to M51 (NGC 5194) as a proof-of-concept, obtaining a resolution of 0.14" (5 pc). Our dust (and gas) surface density map is consistent with independent dust- and CO-based studies at lower resolution. We find that discrepancies between our estimates of surface density and other studies stem primarily from the choice of dust model (i.e. different dust absorption coefficients). When assuming the same dust opacity law, our technique produces surface densities that are consistent with independent studies. This dust extinction technique provides us with gas surface density maps at an unprecedented resolution for full disc coverage studies of nearby galaxies. The resulting well-resolved spatial information opens the possibility for more in-depth examination of the influence of large-scale dynamics (and also stellar feedback mechanisms) on the interstellar medium at parsec-scales, and consequently star formation in nearby galaxies.Comment: MNRAS accepted. 15 pages, 12 figure

Significance of osteopontin expression in human invasive breast tumour stroma

Osteopontin has been reported to stimulate cell adhesion, migration and specific signalling functions. Its overexpression has been found in melanoma, breast, lung, colorectal, stomach and ovarian cancer. However, its overexpression and role in human breast cancer remains to be elucidated. In this study, invasive breast tumours from 129 patients were examined by immunohistochemistry in order to assess osteopontin association with several molecular tumour markers. Additionally, its relationship with proliferation and angiogenesis was determined. Ultimately, other tumour variables such as histological grade, tumour size and nodal status were also assessed. Results achieved showed that no statistical significant association exists between osteopontin expression and major clinicopathological parameters or angiogenesis, except for the number of lymph nodes involved. However, a correlation with some molecular markers was observed, namely with P-Cadherin, EGFR, cytokeratin 14 and vimentin. Additionally, higher proliferation rates were found for the tumours expressing osteopontin. Although several studies refer osteopontin as a potential breast cancer biomarker, it is still not clear if it can provide important diagnosis information, evaluate treatment effects or assess the potential for metastatic disease in patients.Fundação para a Ciência e a Tecnologia (FCT

New method to assess mitophagy flux by flow cytometry

PMID:25945953 WOS:000355323800009publishersversionpublishe

Carbon Monoxide Modulation of Microglia-Neuron Communication: Anti-Neuroinflammatory and Neurotrophic Role

This work was financed by FEDER-Fundo Europeu de Desenvolvimento Regional funds through the COMPETE 2020-Operational Programme for Competitiveness and Internationalization (POCI), Portugal 2020, and by Portuguese funds through FCT-Fundação para a Ciência e a Tecnologia/Ministério da Ciência (FCT), Tecnologia e Ensino Superior in the framework of FCT-ANR/NEU-NMC/0022/2012 grant, PTDC/MEC-NEU/28750/2017 grant, Applied Molecular Biosciences Unit-UCIBIO (UID/Multi/04378/2019) grant; LA/P/0140/2020 of the Associate Laboratory Institute for Health and Bioeconomy; and FCT provided individual financial support to NLS (PD/BD/127819/2016), BFM (PD/BD/128336/2017) and HLAV (IF/00185/2012).Microglia, the ‘resident immunocompetent cells’ of the central nervous system (CNS), are key players in innate immunity, synaptic refinement and homeostasis. Dysfunctional microglia contribute heavily to creating a toxic inflammatory milieu, a driving factor in the pathophysiology of several CNS disorders. Therefore, strategies to modulate the microglial function are required to tackle exacerbated tissue inflammation. Carbon monoxide (CO), an endogenous gaseous molecule produced by the degradation of haem, has anti-inflammatory, anti-apoptotic, and pro-homeostatic and cytoprotective roles, among others. ALF-826A, a novel molybdenum-based CO-releasing molecule, was used for the assessment of neuron-microglia remote communication. Primary cultures of rat microglia and neurons, or the BV-2 microglial and CAD neuronal murine cell lines, were used to study the microglia-neuron interaction. An approach based on microglial-derived conditioned media in neuronal culture was applied. Medium derived from CO-treated microglia provided indirect neuroprotection against inflammation by limiting the lipopolysaccharide (LPS)-induced expression of reactivity markers (CD11b), the production of reactive oxygen species (ROS) and the secretion of inflammatory factors (TNF-α, nitrites). This consequently prevented neuronal cell death and maintained neuronal morphology. In contrast, in the absence of inflammatory stimulus, conditioned media from CO-treated microglia improved neuronal morphological complexity, which is an indirect manner of assessing neuronal function. Likewise, the microglial medium also prevented neuronal cell death induced by pro-oxidant tert-Butyl hydroperoxide (t-BHP). ALF-826 treatment reinforced microglia secretion of Interleukin-10 (IL-10) and adenosine, mediators that may protect against t-BHP stress in this remote communication model. Chemical inhibition of the adenosine receptors A2A and A1 reverted the CO-derived neuroprotective effect, further highlighting a role for CO in regulating neuron-microglia communication via purinergic signalling. Our findings indicate that CO has a modulatory role on microglia-to-neuron communication, promoting neuroprotection in a non-cell autonomous manner. CO enhances the microglial release of neurotrophic factors and blocks exacerbated microglial inflammation. CO improvement of microglial neurotrophism under non-inflammatory conditions is here described for the first time.publishersversionepub_ahead_of_prin

Molecular clouds in M51 from high-resolution extinction mapping

Here, we present the cloud population extracted from M51, following the application of our new high-resolution dust extinction technique to the galaxy. With this technique, we are able to image the gas content of the entire disc of M51 down to 5 pc (0.14 arcsec), which allows us to perform a statistical characterization of well-resolved molecular cloud properties across different large-scale dynamical environments and with galactocentric distance. We find that cloud growth is promoted in regions in the galaxy where shear is minimized; i.e. clouds can grow into higher masses (and surface densities) inside the spiral arms and molecular ring. We do not detect any enhancement of high-mass star formation towards regions favourable to cloud growth, indicating that massive and/or dense clouds are not the sole ingredient for high-mass star formation. We find that in the spiral arms there is a significant decline of cloud surface densities with increasing galactocentric radius, whilst in the inter-arm regions they remain relatively constant. We also find that the surface density distribution for spiral arm clouds has two distinct behaviours in the inner and outer galaxy, with average cloud surface densities at larger galactocentric radii becoming similar to inter-arm clouds. We propose that the tidal interaction between M51 and its companion (NGC 5195) – which heavily affects the nature of the spiral structure – might be the main factor behind this

Neuroprotective effects of digested polyphenols from wild blackberry species

Abstract Purpose Blackberry ingestion has been demonstrated to attenuate brain degenerative processes with the benefits ascribed to the (poly)phenolic components. The aim of this work was to evaluate the neuroprotective potential of two wild blackberry species in a neurodegeneration cell model and compare them with a commercial variety. Methods This work encompasses chemical characterization before and after an in vitro digestion and the assessment of neuroprotection by digested metabolites. Some studies targeting redox/cell death systems were also performed to assess possible neuroprotective molecular mechanisms. Results The three blackberry extracts presented some quantitative differences in polyphenol composition that could be responsible for the different responses in the neurodegeneration cell model. Commercial blackberry extracts were ineffective but both wild blackberries, Rubus brigantinus and Rubus vagabundus, presented neuroprotective effects. It was verified that a diminishment of intracellular ROS levels, modulation of glutathione levels and activation of caspases occurred during treatment. The last effect suggests a preconditioning effect since caspase activation was not accompanied by diminution in cell death and loss of functionality. Conclusions This is the first time that metabolites obtained from an in vitro digested food matrix, and tested at levels approaching the concentrations found in human plasma, have been described as inducing an adaptative response