Optimisation des conditions de culture des cellules endothéliales cornéennes félines pour la reconstruction d'un endothélium cornéen autologue

Les cellules endothéliales cornéennes constituent un tissu à la fois extrêmement particulier et d'une importance singulière. En effet, celui-ci est pourvu d'une capacité régénératrice pratiquement nulle, mais son intégrité est essentielle à la transparence cornéenne et donc à la vision. Par contre, l'endothélium cornéen fait défaut dans diverses pathologies telles que la dystrophie de Fuch et la kératopathie bulleuse du pseudophaque. La méthode traditionnelle pour le remplacement d'un endothélium cornéen pathologique est la greffe de cornée pleine épaisseur. Par contre, la greffe lamellaire postérieure est une alternative chirurgicale en plein essor, très intéressante, puisqu'elle permet de remplacer seulement la portion malade de la cornée. Cette technique chirurgicale a ouvert la porte à une nouvelle possibilité au niveau du génie tissulaire. En effet, il est maintenant envisageable de greffer un endothélium produit en laboratoire. Un défi supplémentaire, que s'est imposé notre équipe, est de reconstruire non seulement un endothélium complet, mais d'en reconstruire un qui serait également autologue, l'immunogénicité jouant un rôle non négligeable lors d'échec de greffes. Mon rôle dans ce projet était donc d'évaluer la faisabilité de reconstruire par génie tissulaire un endothélium cornéen autologue pour notre modèle animal, et ce, à des fins de greffes. D'abord en optimisant les conditions de culture afin de permettre la croissance des cellules endothéliales cornéennes animales. Puis en reconstruisant in vitro un endothélium à partir d'une biopsie cornéenne de taille minimale. Un endothélium sain similaire au tissu natif a ainsi été reconstruit

IPV, distress, and affect dysregulation

Despite an increase in research initiatives and prevention campaigns, intimate partner violence (IPV) remains a public health problem that affects many victims worldwide. The current study aims to examine whether psychological distress symptoms (anger, depression, anxiety) are indirectly related to the perpetration of IPV (physical assault, psychological abuse, coercive control) through affect dysregulation (AD) in men seeking help. Online questionnaires assessing psychological distress symptoms, AD, and violent behaviors were completed by 335 adult men entering treatment for IPV. A path analysis model revealed the indirect associations between psychological distress symptoms and higher IPV perpetration through higher AD. Symptoms of anger were indirectly related to the three forms of perpetrated IPV through higher AD. Symptoms of depression were, directly and indirectly, related to the three forms of perpetrated IPV through higher AD. Finally, symptoms of anxiety were directly related to lower physical assault perpetration, and indirectly related to higher physical assault and coercive control perpetration through higher AD. The final model explained 10% of the variance in perpetrated physical assault, 23% of the variance in perpetrated psychological abuse, and 13% of the variance in perpetrated coercive control. These results underline the necessity of assessing and addressing symptoms of psychological distress and AD among men perpetrators in the treatment of IPV

Attachment, couple communication and sexual coercion

Abstract : The use of subtle strategies to have sex with an unwilling partner is harmful to a couple’s sexual well-being but these strategies remain understudied. This research examined the mediating role of communication patterns in the associations between attachment insecurities and sustained sexual coercion in 145 same- and cross-gender couples, and the moderating role of partners’ gender. In addition to actor and partner effects, results revealed significant indirect effects from attachment insecurities to sexual coercion via communication patterns, with moderating effects of gender. Results may help practitioners and researchers understand the ways attachment insecurities and dysfunctional communication patterns can manifest in the experience of subtle forms of sexual coercion within couples

Impact of cell source on human cornea reconstructed by tissue engineering

Purpose: To investigate the effect of the tissue origin of stromal fibroblasts and epithelial cells on reconstructed corneas in vitro. Methods: Four types of constructs were produced by the self-assembly approach using the following combinations of human cells: corneal fibroblasts/corneal epithelial cells, corneal fibroblasts/skin epithelial cells, skin fibroblasts/corneal epithelial cells, skin fibroblasts/skin epithelial cells. Fibroblasts were cultured with ascorbic acid to produce stromal sheets on which epithelial cells were cultured. After 2 weeks at the air-liquid interface, the reconstructed tissues were photographed, absorption spectra were measured, and tissues were fixed for histologic analysis. Cytokine expression in corneal- or skin-fibroblast-conditioned media was determined with the use of protein array membranes. The effect of culturing reconstructed tissues with conditioned media, or media supplemented with a cytokine secreted mainly by corneal fibroblasts, was determined. Results: The tissue source from which epithelial and mesenchymal cells were isolated had a great impact on the macroscopic and histologic features (epithelium thickness and differentiation) and the functional properties (transparency) of the reconstructed tissues. The reconstructed cornea had ultraviolet-absorption characteristics resembling those of native human cornea. The regulation of epithelial differentiation and thickness was mesenchyme-dependent and mediated by diffusible factors. IL-6, which is secreted in greater amounts by corneal fibroblasts than skin fibroblasts, decreased the expression of the differentiation marker DLK in the reconstructed epidermis. Conclusions: The tissue origin of fibroblasts and epithelial cells plays a significant role in the properties of the reconstructed tissues. These human models are promising tools for gaining a thorough understanding of epithelial-stromal interactions and regulation of epithelia homeostasis

Reconstruction of a human cornea by the self-assembly approach of tissue engineering using the three native cell types

Purpose: The purpose of this study was to produce and characterize human tissue-engineered corneas reconstructed using all three corneal cell types (epithelial, stromal, and endothelial cells) by the self-assembly approach. Methods: Fibroblasts cultured in medium containing serum and ascorbic acid secreted their own extracellular matrix and formed sheets that were superposed to reconstruct a stromal tissue. Endothelial and epithelial cells were seeded on each side of the reconstructed stroma. After culturing at the air-liquid interface, the engineered corneas were fixed for histology and transmission electron microscopy (TEM). Immunofluorescence labeling of epithelial keratins, basement membrane components, Na+/K+-ATPase α1, and collagen type I was also performed. Results: Epithelial and endothelial cells adhered to the reconstructed stroma. After 10 days at the air-liquid interface, the corneal epithelial cells stratified (4 to 5 cell layers) and differentiated into well defined basal and wing cells that also expressed Na+/K+-ATPase α1 protein, keratin 3/12, and basic keratins. Basal epithelial cells from the reconstructed epithelium formed many hemidesmosomes and secreted a well defined basement membrane rich in laminin V and collagen VII. Endothelial cells formed a monolayer of tightly-packed cells and also expressed the function related protein Na+/K +-ATPase α1. Conclusions: This study demonstrates the feasibility of producing a complete tissue-engineered human cornea, similar to native corneas, using untransformed fibroblasts, epithelial and endothelial cells, without the need for exogenous biomaterial

Internal deformation of the subducted Nazca slab inferred from seismic anisotropy

Within oceanic lithosphere a fossilized fabric is often preserved originating from the time of plate formation. Such fabric is thought to form at the mid-ocean ridge when olivine crystals align with the direction of plate spreading1, 2. It is unclear, however, whether this fossil fabric is preserved within slabs during subduction or overprinted by subduction-induced deformation. The alignment of olivine crystals, such as within fossil fabrics, can generate anisotropy that is sensed by passing seismic waves. Seismic anisotropy is therefore a useful tool for investigating the dynamics of subduction zones, but it has so far proved difficult to observe the anisotropic properties of the subducted slab itself. Here we analyse seismic anisotropy in the subducted Nazca slab beneath Peru and find that the fast direction of seismic wave propagation aligns with the contours of the slab. We use numerical modelling to simulate the olivine fabric created at the mid-ocean ridge, but find it is inconsistent with our observations of seismic anisotropy in the subducted Nazca slab. Instead we find that an orientation of the olivine crystal fast axes aligned parallel to the strike of the slab provides the best fit, consistent with along-strike extension induced by flattening of the slab during subduction (A. Kumar et al., manuscript in preparation). We conclude that the fossil fabric has been overprinted during subduction and that the Nazca slab must therefore be sufficiently weak to undergo internal deformation

Using gene expression to identify the most suitable environmental conditions for juvenile deepwater redfish (Sebastes mentella) growth and metabolism in the Estuary and the Gulf of St. Lawrence.

International audienceAbstract Deepwater redfish Sebastes mentella will be among the most important resource-sustaining commercial bottom-fish fisheries in the years to come in the Estuary and Gulf of St. Lawrence (EGSL). In 2011, 2012, and 2013, three strong cohorts were recruited to the stock; their abundance in 2018 was 80 times higher than that of the 1993–2012 period. The main goal of this work was to deepen our knowledge of their growth regulation and metabolism in order to identify molecular indicators and determine how they are influenced by natural environmental conditions. Fish weight and water temperature explained 11% of the variation in relative mRNA levels of specific gene targets in liver and muscle among seven sites where deepwater redfish were captured in the EGSL. The relative expression of liver insulin-like growth factor-1 (igf-1) and white muscle A-chain lactate dehydrogenase (ldh-a) correlate positively with weight, whereas heavy chain muscle myosin (myo), heart citrate synthase (cs), and white muscle pyruvate kinase (pk) correlate negatively. The relative expression of heart cytochrome c oxidase subunit 1 (cox-1) and white muscle igf-1 receptor isoform a (igf-1ra) correlate negatively with temperature. Deepwater redfish from the estuary were smaller than those caught at other sites. Since the growth potential of deepwater redfish was strongly correlated with temperature (being enhanced by higher temperatures), this study suggests an ecological advantage for this species in a climate-warming context

Décolonisation humanitaire et transformations structurelles

The present article outlines the internationalization strategy of the Center for International Studies and Cooperation (CECI) developed within the context of Canadian policy and international normative frameworks that call for the strengthening and empowerment of ecosystems. The results of this collaborative research, conducted using a mixed-methods methodology, reveal the organizational drivers and barriers within CECI’s transformation strategy. Our analysis, rooted in the work of critical humanitarian action authors within NGOs, enriches Permultter’s model by highlighting key transition moments during the implementation of CECI’s organizational strategy to decolonize its network

Regional variation of gene regulation associated with storage lipid metabolism in American glass eels (Anguilla rostrata)

Variation in gene regulation may be involved in the differences observed for life history traits within species. American eel (Anguilla rostrata) is well known to harbor distinct ecotypes within a single panmictic population. We examined the expression of genes involved in the regulation of appetite as well as lipid and glycogen among glass eels migrating to different locations on the Canadian east coast and captured at two different periods of upstream migration. Gene expression levels of three reference and five candidate genes were analyzed by real-time PCR with Taqman probes in recently captured wild glass eels. All gene transcripts were detected in glass eels. Of the five candidate genes, bile salt activated and triacylglycerol lipases were respectively 7.65 and 3.25 times more expressed in glass eels from the St. Lawrence estuary than in those from Nova Scotia, and there was no effect related to the two-week difference in capture date. These two genes explained 82.41% of the dissimilarity between the two rivers. In contrast, glycogen phosphorylase, ghrelin, and leptin receptor genes showed no significant differences in gene transcription. These results confirmed at the molecular level an observation that was recently made at the phenotypic level that glass eels from the St. Lawrence estuary have a greater capacity to use lipid reserves to sustain their metabolic needs. These observations add to the body of evidence supporting the hypothesis that regional phenotypic variation observed in American eel is determined early in life and that part of this variation is likely under genetic control. -- Keywords : Ecotypes ; Ghrelin ; Glycogen phosphorylase ; Leptin receptor ; Lipase ; mRNA expression ; Transcriptomics