Microglial cells in astroglial cultures: a cautionary note

Primary rodent astroglial-enriched cultures are the most popular model to study astroglial biology in vitro. From the original methods described in the 1970's a great number of minor modifications have been incorporated into these protocols by different laboratories. These protocols result in cultures in which the astrocyte is the predominant cell type, but astrocytes are never 100% of cells in these preparations. The aim of this review is to bring attention to the presence of microglia in astroglial cultures because, in my opinion, the proportion of and the role that microglial cells play in astroglial cultures are often underestimated. The main problem with ignoring microglia in these cultures is that relatively minor amounts of microglia can be responsible for effects observed on cultures in which the astrocyte is the most abundant cell type. If the relative contributions of astrocytes and microglia are not properly assessed an observed effect can be erroneously attributed to the astrocytes. In order to illustrate this point the case of NO production in activated astroglial-enriched cultures is examined. Lipopolysaccharide (LPS) induces nitric oxide (NO) production in astroglial-enriched cultures and this effect is very often attributed to astrocytes. However, a careful review of the published data suggests that LPS-induced NO production in rodent astroglial-enriched cultures is likely to be mainly microglial in origin. This review considers cell culture protocol factors that can affect the proportion of microglial cells in astroglial cultures, strategies to minimize the proportion of microglia in these cultures, and specific markers that allow the determination of such microglial proportions.

In vitro and in vivo ocular biocompatibility of electrospun poly(ɛ-caprolactone) nanofibers.

Biocompatibility is a requirement for the development of nanofibers for ophthalmic applications. In this study, nanofibers were elaborated using poly(ε-caprolactone) via electrospinning. The ocular biocompatibility of this material was investigated. MIO-M1 and ARPE-19 cell cultures were incubated with nanofibers and cellular responses were monitored by viability and morphology. The in vitro biocompatibility revealed that the nanofibers were not cytotoxic to the ocular cells. These cells exposed to the nanofibers proliferated and formed an organized monolayer. ARPE-19 and MIO-M1 cells were capable of expressing GFAP, respectively, demonstrating their functionality. Nanofibers were inserted into the vitreous cavity of the rat's eye for 10days and the in vivo biocompatibility was investigated using Optical Coherence Tomography (OCT), histology and measuring the expression of pro-inflammatory genes (IL-1β, TNF-α, VEGF and iNOS) (real-time PCR). The OCT and the histological analyzes exhibited the preserved architecture of the tissues of the eye. The biomaterial did not elicit an inflammatory reaction and pro-inflammatory cytokines were not expressed by the retinal cells, and the other posterior tissues of the eye. Results from the biocompatibility studies indicated that the nanofibers exhibited a high degree of cellular biocompatibility and short-term intraocular tolerance, indicating that they might be applied as drug carrier for ophthalmic use.

Cell type-specific expression and localization of cytochrome P450 isoforms in tridimensional aggregating rat brain cell cultures.

Within the Predict-IV FP7 project a strategy for measurement of in vitro biokinetics was developed, requiring the characterization of the cellular model used, especially regarding biotransformation, which frequently depends on cytochrome P450 (CYP) activity. The extrahepatic in situ CYP-mediated metabolism is especially relevant in target organ toxicity. In this study, the constitutive mRNA levels and protein localization of different CYP isoforms were investigated in 3D aggregating brain cell cultures. CYP1A1, CYP2B1/B2, CYP2D2/4, CYP2E1 and CYP3A were expressed; CYP1A1 and 2B1 represented almost 80% of the total mRNA content. Double-immunolabeling revealed their presence in astrocytes, in neurons, and to a minor extent in oligodendrocytes, confirming the cell-specific localization of CYPs in the brain. These results together with the recently reported formation of an amiodarone metabolite following repeated exposure suggest that this cell culture system possesses some metabolic potential, most likely contributing to its high performance in neurotoxicological studies and support the use of this model in studying brain neurotoxicity involving mechanisms of toxication/detoxication.

Identitat diversa. Conflicte identitari en persones vinculades a diferents cultures

Aquest treball de Final de Grau es centra en investigar un fenomen personal i social experimentat per persones d’origen immigrat al plantejar-se la seva identitat vinculada a dues o més cultures. Aquesta recerca pretén apropar-se a les vivències, pensaments, sentiments, etc. de persones que han viscut un conflicte identitari i esbrinar si han experimentat una certa pressió social per definir la seva identitat en una o altra cultura i com ho han viscut. La investigació compta amb la participació de sis persones d’origen marroquí, algunes nascudes al territori i altres no, les quals expliquen las seva experiència i perspectiva sobre aquest fenomen.

Cyclosporine A kinetics in brain cell cultures and its potential of crossing the blood-brain barrier.

There is an increasing need to develop improved systems for predicting the safety of xenobiotics. However, to move beyond hazard identification the available concentration of the test compounds needs to be incorporated. In this study cyclosporine A (CsA) was used as a model compound to assess the kinetic profiles in two rodent brain cell cultures after single and repeated exposures. CsA induced-cyclophilin B (Cyp-B) secretion was also determined as CsA-specific pharmacodynamic endpoint. Since CsA is a potent p-glycoprotein substrate, the ability of this compound to cross the blood-brain barrier (BBB) was also investigated using an in vitro bovine model with repeated exposures up to 14days. Finally, CsA uptake mechanisms were studied using a parallel artificial membrane assay (PAMPA) in combination with a Caco-2 model. Kinetic results indicate a low intracellular CsA uptake, with no marked bioaccumulation or biotransformation. In addition, only low CsA amounts crossed the BBB. PAMPA and Caco-2 experiments revealed that CsA is mostly trapped to lipophilic compartments and exits the cell apically via active transport. Thus, although CsA is unlikely to enter the brain at cytotoxic concentrations, it may cause alterations in electrical activity and is likely to increase the CNS concentration of other compounds by occupying the BBBs extrusion capacity. Such an integrated testing system, incorporating BBB, brain culture models and kinetics could be applied for assessing neurotoxicity potential of compounds.

Amiodarone biokinetics, the formation of its major oxidative metabolite and neurotoxicity after acute and repeated exposure of brain cell cultures.

The difficulty in mimicking nervous system complexity and cell-cell interactions as well as the lack of kinetics information has limited the use of in vitro neurotoxicity data. Here, we assessed the biokinetic profile as well as the neurotoxicity of Amiodarone after acute and repeated exposure in two advanced rodent brain cell culture models, consisting of both neurons and glial cells organized in 2 or 3 dimensions to mimic the brain histiotypic structure and function. A strategy was applied to evidence the abiotic processes possibly affecting Amiodarone in vitro bioavailability, showing its ability to adsorb to the plastic devices. At clinically relevant Amiodarone concentrations, known to induce neurotoxicity in some patients during therapeutic treatment, a complete uptake was observed in both models in 24h, after single exposure. After repeated treatments, bioaccumulation was observed, especially in the 3D cell model, together with a greater alteration of neurotoxicity markers. After 14days, Amiodarone major oxidative metabolite (mono-N-desethylamiodarone) was detected at limited levels, indicating the presence of active drug metabolism enzymes (i.e. cytochrome P450) in both models. The assessment of biokinetics provides useful information on the relevance of in vitro toxicity data and should be considered in the design of an Integrated Testing Strategy aimed to identify specific neurotoxic alerts, and to improve the neurotoxicity assay predictivity for human acute and repeated exposure.

Courts and Courtly Cultures in Early Modern Italy and Europe: Models and Languages

This volume deals with the forms of propaganda and self-representation, through words and images, during the rise of the 'civiltà delle corti' and through processes typical of the time, such as confrontation, adaptation, competition and rivalry. This period, which marked the passage of Italian and European culture from the Middle Ages to the Renaissance, is fundamental in the development of Modern Europe, and it lasted up to the XVIII century and beyond. At the heart of many matters debated here lies the relationship between culture and politics. The formation of a 'Lombard identity', central to the Sinergia project which was the frame of the whole research and its conferences, is closely linked to this broad general context. It places the so called 'questione milanese' - above the traditional hierarchies 'Toscana oriented' - at the centre of many questions regarding Northern Italy as a whole, starting from the dissolution of the Medieval communes, through to the rise of the signorie, from the end of the XIII century and the beginning of the XIV century up to the early XVI century.

Identification des dermatophytes en cultures sur la base de leurs critères macroscopiques et microscopiques

Contexte : Les dermatophytes sont des champignons filamenteux parasites spécialisés qui dégradent les tissus kératinisés. Ils sont responsables de la plupart des mycoses de la peau, du cuir chevelu et des cheveux, et des ongles. Le choix du traitement des dermatophytoses dépend des symptômes et du dermatophyte incriminé parmi une quinzaine d'espèces possibles. L'identification des dermatophytes se fait en général sur la base des caractères macroscopiques et microscopiques des cultures. L'identification est parfois difficile ou reste incertaine car il peut y avoir des variations d'un isolat à l'autre au sein d'une même espèce. Cependant, les espèces sont facilement identifiées sur la base de séquences d'ADN. En pratique, des séquences d'ADN ribosomique suffisamment polymorphes sont le plus souvent utilisées pour discriminer les espèces de dermatophytes. Des méthodes spécialisées et sophistiquées telles que les séquences d'ADN et la spectrométrie de masse sont de plus en plus proposées dans la littérature pour identifier les dermatophytes. Toutefois, ces méthodes ne peuvent pas être utilisées directement par un médecin dans un cabinet médical. C'est pourquoi des méthodes plus simples basées sur l'observation de caractères phénotypiques des champignons en culture ne devraient pas être abandonnées. Objectif : Etablir une clé d'identification dichotomique se basant sur des caractères macroscopiques et microscopiques permettant une identification fiable du dermatophyte par la culture. Des clés d'identification des espèces seront élaborées et testées pour leur validation en parallèle avec leur identification par des méthodes de Biologie Moléculaire. Créer un outil simple qui pourra être utilisé au laboratoire par des médecins ou des biologistes non spécialisés en mycologie pour identifier les dermatophytes sans avoir recours à une technologie sophistiquée. Méthodes : Inventaire des espèces isolées de 2001 à 2012 au laboratoire de dermatologie du CHUV. Inventaire des caractères phénotypiques permettant de caractériser chaque espèce. Création d'un système dichotomique sur la base des caractères phénotypiques pour séparer et identifier les espèces (clé d'identification des espèces). Résultats attendus : Les résultats attendus sont définis au niveau des objectifs. L'outil doit être accessible pour des personnes inexpérimentées qui pourront alors identifier les dermatophytes. Plus-value : Les dermatophytoses sont fréquemment diagnostiquées. Cet outil est destiné à tous les dermatologues installés et au personnel de laboratoire qui ne sont pas nécessairement spécialisés en la matière.

Faire fondre la banquise : la difficile ouverture des villes suisses aux cultures jeunes

L'année 1980 représente une année charnière pour le développement des grandes villes de Suisse. Jusqu'alors marquées par une offre culturelle extrêmement pauvre et peu diversifiée, l'apparition et l'activité des scènes musicales jeunes vont les obliger à changer de politique. Cette arrivée soudaine des scènes de musique va d'abord passer par une phase conflictuelle. Pour faire fondre la banquise conservatrice qui recouvre alors les différentes villes, les jeunes acteurs de ces scènes vont devoir se mobiliser pour conquérir des territoires urbains. Mais par la suite, les autorités vont engager des réformes prenant en compte les revendications des scènes musicales quant à leur existence dans le paysage urbain. En s'intéressant aux cinq plus grandes villes de Suisse (Zurich, Bâle, Genève, Berne et Lausanne), cet article souhaite démontrer comment les scènes musicales ont permis de moderniser ces villes, en en renouvelant l'identité et en s'inscrivant concrètement dans leur territoire. Dans une perspective socio-historique, cette étude croisera analyse d'archives, témoignages d'individus membres de ces scènes, données quantitatives sur l'offre culturelle et études des politiques gouvernementales. Il s'agira à la fois d'observer l'impact concret des scènes musicales sur ces villes et de se demander si cette intégration implique des formes de récupération ou de transformation de ce ses scènes.

Critical behavior of a water monolayer under hydrophobic confinement

The properties of water can have a strong dependence on the confinement. Here, we consider a water monolayer nanoconfined between hydrophobic parallel walls under conditions that prevent its crystallization. We investigate, by simulations of a many-body coarse-grained water model, how the properties of the liquid are affected by the confinement. We show, by studying the response functions and the correlation length and by performing finite-size scaling of the appropriate order parameter, that at low temperature the monolayer undergoes a liquid-liquid phase transition ending in a critical point in the universality class of the two-dimensional (2D) Ising model. Surprisingly, by reducing the linear size L of the walls, keeping the walls separation h constant, we find a 2D-3D crossover for the universality class of the liquid-liquid critical point for L/h=~50, i.e. for a monolayer thickness that is small compared to its extension. This result is drastically different from what is reported for simple liquids, where the crossover occurs for , and is consistent with experimental results and atomistic simulations. We shed light on these findings showing that they are a consequence of the strong cooperativity and the low coordination number of the hydrogen bond network that characterizes water.

Enhanced triterpene production in Tabernaemontana catharinensis cell suspension cultures in response to biotic elicitors

Cell suspension cultures of Tabernaemontana catharinensis were treated with autoclaved homogenates of Candida albicans, Fusarium oxysporum, Penicillium avelanium and Saccharomyces cerevisiae. The effects caused by the concentration, exposure time and the type of elicitor on the accumulation of pentacyclic triterpenes were monitored. When exposed to biotic elicitors for longer periods, some cell lines redoubled the production of those triterpenes. Saccharomyces cerevisiae homogenate was the best elicitor of triterpenes in all cell lines investigated.

Les cultures en contacte

L’evidència de la multiplicitat de cultures a l’escola planteja molts interrogants sobre la manera com han de ser-hi presents aquestes cultures. Si volem que contribueixin a construir una cultura comuna plenament inserida en el nostre país, com podem convertir les dificultats d’aquest objectiu en clau d’oportunitat?

Leaf and root volatiles produced by tissue cultures of Alpinia zerumbet (pers.) Burtt & Smith under the influence of different plant growth regulators

Volatiles produced by plantlets of Alpinia zerumbet were obtained by means of simultaneous distillation-extraction (SDE). The effects of indole-3-acetic acid, kinetin, thidiazuron and 6-benzylaminopurine on leaf and root volatile composition obtained by tissue cultures were investigated. A higher content of b-pinene and a lower content of sabinene were observed in leaf volatile of plantlets cultured in control, IAA and IAA+ TDZ media, as compared with those of donor plants. In vitro conditions were favorable to increase caryophyllene content. Volatile compounds from the root were characterized mainly by camphene, fenchyl-acetate and bornyl acetate; which constitute about 60% of total volatile.

Interaction between cationic and conventional nonionic surfactants in the mixed micelle and monolayer formed in aqueous medium

Mixed micellization and surface properties of cationic and nonionic surfactants dimethyl decyl-, tetradecyl- and hexadecyl phosphineoxide mixtures are studied using conductivity and surface tension measurements. The models of Rubingh, Rosen, and Clint, are used to obtain the interaction parameter, minimum area per molecule, mixed micelle composition, free energies of mixing and activity coefficients. The micellar mole fractions were always higher than ideal values indicating high contributions of cationics in mixed micelles. Activity coefficients were less than unity indicating synergism in micelles. The negative free energies of mixing showed the stability of the surfactants in the mixed micelles.