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See detailOutils d’aide à la réussite dans le cadre du cours de Biologie en Premier bachelier en Faculté des Sciences à l’Université de Liège : présentation et analyse
Thiry, Marc ULg; Rigo, Pierre ULg; Poffé, Corentin ULg

Conference (2015, August 26)

A l’Université de Liège, les cours du premier quadrimestre sont communs à certaines sections du premier bachelier de la Faculté des Sciences. Le programme de ce quadrimestre commun reprend un cours de ... [more ▼]

A l’Université de Liège, les cours du premier quadrimestre sont communs à certaines sections du premier bachelier de la Faculté des Sciences. Le programme de ce quadrimestre commun reprend un cours de biologie. Depuis plusieurs années, le titulaire de ce cours de biologie met en place diverses actions visant à favoriser la transition secondaire-université : séminaires, remédiations, aides à l’étude, forum, livre de référence, etc. Ces différentes actions font l’objet d’un suivi didactique, permettant un monitoring précis et une régulation au fil du temps. Dans un premier temps, la communication présentera l’état des lieux des outils mis à la disposition des étudiants en vue de favoriser leur réussite pour le cours de biologie, mais aussi pour les autres cours, par transfert des compétences acquises. Dans un second temps, le dispositif de suivi didactique des séances d'aide à l'étude et ses principaux résultats seront présentés. [less ▲]

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See detailSpatio‑temporal dynamics of β‑tubulin isotypes during the development of the sensory auditory organ in rat
Renauld, Justine ULg; Johnen, Nicolas; Thelen, Nicolas ULg et al

in Histochemistry & Cell Biology (2015)

There are different β-tubulin isoforms in microtubules of vertebrate tissues. However, their functional significance is still largely unknown. In the present study, we investigated the localization of ... [more ▼]

There are different β-tubulin isoforms in microtubules of vertebrate tissues. However, their functional significance is still largely unknown. In the present study, we investigated the localization of five β-tubulin isotypes (β1– 5) within the hearing organ during development in rat. By using confocal microscopy, we showed that with the exception of the β3-tubulin isoform that was specific to nerve fibres, all the different β-tubulin isoforms were mainly present in the supporting cells. Contrary to β1–4-tubulins, we also found that the β5-tubulin isoform appeared only at a key stage of the post-natal development in specific cell types (pillar cells and Deiters’ cells). By using transmission electron microscopy, we revealed further that this developmental stage coincided with the formation of two separate bundles of microtubules from a unique one in these supporting cells. Together, these results suggest that the β5-tubulin isoform might be involved in the generation of new microtubule bundles from a pre-existing one. [less ▲]

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See detailIdentification of VZV ORF9p potential cellular partners that could be important for the viral egress.
Lebrun, Marielle ULg; riva, laura; Rambout, Xavier ULg et al

Poster (2015, July 26)

ORF9p (homologous to HSV-1 VP22) is a VZV tegument protein essential for the viral replication. During the lytic cycle it is the mostly expressed gene. We have recently demonstrated that it is a substrate ... [more ▼]

ORF9p (homologous to HSV-1 VP22) is a VZV tegument protein essential for the viral replication. During the lytic cycle it is the mostly expressed gene. We have recently demonstrated that it is a substrate of the viral kinase ORF47p and that its ORF47p-dependent phosphorylation is important for the secondary envelopment process. We also have identified an acidic cluster (AC) within the protein that is important for its correct localization in the infected cells and for the interaction with ORF47p. The recombinant VZV expressing ORF9p-ΔAC presents an accumulation of capsids in the perinuclear space. ORF9p seems then to play an important role in several steps of the egress process. In this context, we sought to identify cellular partners of ORF9p that might be important for these functions. We performed a yeast two hybrid screen against the human ORFeome 5.1. and picked out 44 candidates among which 5 proteins playing roles in membrane organization and targeting. We currently are trying to confirm these interactions in infected cells and to assess the role of these interactions for the viral lytic cycle. [less ▲]

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See detailDUSP3 Phosphatase Deficiency or Inhibition Limits Platelet Activation and Arterial Thrombosis
Rahmouni, Souad ULg; MUSUMECI, Lucia ULg; Kuijpers, Marijke J et al

Conference (2015, June 24)

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See detailSpatio-temporal dynamics of β-tubulin isotypes and acetyl-alpha tubulin during the development of the sensory auditory organ in rodents.
Renauld, Justine ULg; Thelen, Nicolas ULg; Johnen, Nicolas ULg et al

Poster (2015, June 05)

The auditory organ is a highly specialized structure composed by specific cellular types. The sensory cells are characterized by stereocilia at their apex and are necessary for the sound perception. These ... [more ▼]

The auditory organ is a highly specialized structure composed by specific cellular types. The sensory cells are characterized by stereocilia at their apex and are necessary for the sound perception. These cells are supported by supporting cells. Supporting cells possess a characteristic cytoskeleton in direct relation with their morphological features and their development. There are different β-tubulin isoforms in microtubules of vertebrate tissues. However, their functional significance is still largely unknown. In the present study, we investigated the localization of five β-tubulin isotypes (β 1 to 5) as well as acetyl-α-tubulin within the hearing organ during development in rodents. By using confocal microscopy, we showed that with the exception of the β3-tubulin isoform that was specific to nerve fibres, all the different β-tubulin isoforms and acetyl-α-tubulin were mainly present in the supporting cells. Contrary to β1-4-tubulins, we also found that the β5-tubulin isoform appeared only at a key stage of the postnatal development in specific cell types (pillar cells and Deiters’ cells). By using transmission electron microscopy, we revealed further that this developmental stage coincided with the formation of two separate bundles of microtubules from a unique one in these supporting cells. In conclusion, these results suggest that the β5-tubulin isoform might be involved in the generation of new microtubule bundles from a pre-existing one. [less ▲]

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See detailVaricella-Zoster Virus (VZV) assemblons interplay with PML bodies
Lebrun, Marielle ULg; Thiry, Marc ULg; Sadzot, Catherine ULg

Poster (2015, May 15)

Using a virus expressing the small capsid protein fused to an eGFP tag (eGFP-ORF23 VZV), we recently identified, in the nuclei of infected cells, the presence of dynamic capsid aggegrates. Because we ... [more ▼]

Using a virus expressing the small capsid protein fused to an eGFP tag (eGFP-ORF23 VZV), we recently identified, in the nuclei of infected cells, the presence of dynamic capsid aggegrates. Because we believe that these structures might represent sites of preferential caspid assembly and by analogy with HSV-1, we referred to them as “VZV assemblons”. Structures resembling these assemblons and identified as capsids entrapped in some “PML-cages” were recently described in the nuclei of wild-type VZV infected cells (Reichelt et al., 2011). We then wonder if there was a link between these independent observations. When we infected MeWo cells in which the expression of each PML subunit is downregulated by shRNA, VZV assemblons still formed. Immunostaining of MeWo cells infected by eGFP-ORF23 VZV with an antibody against the PML protein showed that VZV assemblons only partially colocalize with PML bodies. However, overexpression of PML-I-eGFP in HEK293 cells followed by infection with a tagRFP-T-ORF23 VZV, where the ORF23 protein is fused to a red tag, showed a complete colocalization is complete. The same result was obtained with all tested PML isoforms. This suggests that the partial colocalization in normal cells could be due to the expression level of PML proteins. Altogether, these results suggest that rather than a progressive accumulation of newly formed capsids within PML cages, it is likely that PML protein is recruited to the sites where VZV assemblons develop. It correlates with the fact that the number of PML bodies decreases with the infection. Obviously, even if this phenomenon might impede the egress of a substantial amount of capsids and, in this regard, limit the infection progression, all the tested cell lines are permissive to VZV. It would then be interesting to investigate the relationship between VZV assemblons and PML bodies in latent or non permissive VZV infection models. [less ▲]

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See detailDUSP3 Phosphatase Deficiency or Inhibition Limit Platelet Activation and Arterial Thrombosis
Musumeci, Lucia ULg; Kuijpers, Marijke; Gilio, Karen et al

in Circulation (2015), 131(7), 656-68

Background A limitation of current antiplatelet therapies is their inability to separate thrombotic events from bleeding occurrences. Better understanding of the molecular mechanisms leading to platelet ... [more ▼]

Background A limitation of current antiplatelet therapies is their inability to separate thrombotic events from bleeding occurrences. Better understanding of the molecular mechanisms leading to platelet activation is of importance for the development of improved therapies. Recently, protein tyrosine phosphatases (PTPs) have emerged as critical regulators of platelet function. Methods and Results This is the first report implicating the dual-specificity phosphatase 3 (DUSP3) in platelet signaling and thrombosis. This phosphatase is highly expressed in human and mouse platelets. Platelets from DUSP3-deficient mice displayed a selective impairment of aggregation and granule secretion mediated through the collagen receptor glycoprotein VI (GPVI) and the C-type lectin-like receptor 2 (CLEC-2). DUSP3-deficient mice were more resistant to collagen- and epinephrine-induced thromboembolism, compared to wild-type mice, and showed severely impaired thrombus formation upon ferric chloride-induced carotid artery injury. Intriguingly, bleeding times were not altered in DUSP3-deficient mice. At the molecular level, DUSP3 deficiency impaired Syk tyrosine phosphorylation, subsequently reducing phosphorylation of PLCγ2 and calcium fluxes. To investigate DUSP3 function in human platelets, a novel small-molecule inhibitor of DUSP3 was developed. This compound specifically inhibited collagen and CLEC-2-induced human platelet aggregation, thereby phenocopying the effect of DUSP3 deficiency in murine cells. Conclusions DUSP3 plays a selective and essential role in collagen- and CLEC-2-mediated platelet activation and thrombus formation in vivo. Inhibition of DUSP3 may prove therapeutic for arterial thrombosis. This is the first time a PTP, implicated in platelet signaling, has been targeted with a small-molecule drug. [less ▲]

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See detailMyoferlin: an indispensable component in VEGFA secretion by pancreas cancer cells
Fahmy, Karim ULg; Peulen, Olivier ULg; Castronovo, Vincenzo ULg et al

Poster (2015, January 31)

In this poster, our laboratory showed the importance of myoferlin, a biomarker of pancreas cancer, in the controle of VEGF-A mediated angiogenesis. Our laboratory showed that silencing myoferlin in ... [more ▼]

In this poster, our laboratory showed the importance of myoferlin, a biomarker of pancreas cancer, in the controle of VEGF-A mediated angiogenesis. Our laboratory showed that silencing myoferlin in pancreas cancer cells, BxPC-3, provoques a decrease in cell prolifération in vitro and a decrease in tumor volumes in animal model. Myoferlin silencing also provokes a decrease in VEGF-A secretion in the conditioned medium and that decrease was abserved in the animal model as a decrease in microvessels dencity. It appeared that this decrease in secretion is due to a a blockage in the exocytosis. Our data also showed a significate correlation between myoferlin expression and microvessels density in patients section. [less ▲]

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See detailMyoferlin Regulates Endosomal Trafficking and Tunes Cancer Cell Metabolism
Blomme, Arnaud; Costanza, Brunella; Thiry, Marc ULg et al

Poster (2015, January 31)

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See detailBacterial DNA mimetics activate platelets and promote thrombosis via CLEC-2
Delierneux, Céline; Hego, Alexandre; Lecut, Christelle et al

Poster (2015, January 27)

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See detailAccumulation of methylglyoxal, a glycolysis by-product modulates YAP1 transcription co-factor localization and activity in human breast cancer cells through HSP90 modification
Nokin, Marie-Julie; Durieux, Florence; Peixoto, Paul et al

Poster (2015, January 27)

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See detailMyoferlin: an indispensable component in VEGR-A secretion by pancreatic cancer cell
Fahmy, Karim; Gonzalez, Arnaud; Arafad, Mohamed et al

Poster (2015, January 27)

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See detailMyoferlin regulates endosomal trafficking and tunes cancer cell metabolism
Blomme, Arnaud; Costanza, Brunella; Thiry, Marc ULg et al

Poster (2015, January 27)

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See detailLocalization of rDNA transcription sites within reptilian nucleoli
Bartholomé, Odile ULg; Franck, Claire; Thiry, Marc ULg

Poster (2015, January 27)

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See detailChange in the protofilamentnumber and β5-tubulin appearance in supporting cells during development of the hearing organ.
Renauld, Justine ULg; Thelen, Nicolas ULg; Johnen, Nicolas ULg et al

Poster (2015, January)

The supporting cells of the hearing organ are characterized by the presence of an abundant cytoskeleton which is mainly composed of microtubules. These supporting cells have also been shown to contain a ... [more ▼]

The supporting cells of the hearing organ are characterized by the presence of an abundant cytoskeleton which is mainly composed of microtubules. These supporting cells have also been shown to contain a minor mammalian tubulin, the β5-tubulin, recently reported as a biomarker of cell proliferation. It was shown that a β-tubulin isoform can specify the microtubule architecture, as seen with the expression of the Moth β2 tubulin in the Drosophila testes which imposes the 16-protofilament (16pf) structure on the corresponding subset of Drosophila microtubules. Moreover, supporting cell microtubules are formed by 15pf instead of the canonical 13, a unique fact among vertebrates. Such a protofilament configuration has been observed in C. elegans’ neurons which are responsible for the mechanosensory sense of touch. It was also shown that these 15pf microtubules were essential to the proper functioning of these neurons. To determine the role of this particular tubulin in the auditory organ and its possible involvement in the formation of the unusual 15pf microtubules of supporting cells, we studied the spatiotemporal localization of β5-tubulin during development in rats from embryonic day 18 until P25 (25th postnatal day). Then we examined the fine structure of microtubules at the transmission electron microscope level (TEM). Our results showed that β5-tubulin, contrary to other β-tubulins, had a unique distribution in the cochlea. This β-tubulin appeared at a postnatal stage, before the opening of the Corti’s tunnel and is restricted to supporting cells, especially in pillar and Deiters’ cells. Our TEM study further indicated that these cells were composed by 13pf microtubules at P2, but by 15pf microtubules at P25. In conclusion, the architecture and composition of microtubules present in the supporting cells change during development of the Corti organ. Further experiments are now required to determine if these changes are related to the appearance of β5-tubulin. [less ▲]

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