NCube/papyrus huggingface dataset:
Développement d'un biosenseur BRET permettant le criblage de drogues qui causent l'activation de canaux Kir3 via les récepteurs couplés aux protéines G

G protein-coupled receptors form multimeric complexes comprising G protein and effectors. We want to characterize such complexes comprising delta opioid receptors (DOR) and Kir3 channels, which interest us due to their involvement in opioid analgesia. Immunopurification, BRET and GTPgS binding experiments were done in transfected HEK293 cells. Kir3 channels were co-immunopurified with DOR, implying a spontaneous interaction between the receptor and effector. BRET assays corroborated the presence of this interaction in living cells and allowed us to identify a spontaneous and specific interaction between DOR/Gg and Gg/Kir3, indicating their co-existence within the same complex. Since the activation of the receptor implies it undergoes conformational changes, we were interested in evaluating if the conformational information flows from the ligand-bound receptor until the downstream effector. Hence, we determined the effect of different ligands on the BRET signal that was generated by the following pairs: DOR/Gbg, DOR/Kir3 and Kir3/Gbg. We noticed a modulation of the DOR/Gbg and Gbg/Kir3 interactions that followed the order of efficacy of the ligands to activate the G protein, which we did not observe between DOR and Kir3. Therefore, we concluded that the conformational information flows from the receptor to the Kir3 channel via the Gbg protein. These results allowed us to develop a BRET biosensor (EYFP-Gg2/Kir3.1-Rluc), which could be used in high throughput screening to detect new molecules that activate Kir3 channels with high efficacy.

Keyphrases: HEK293, conformations multiples, multimeric complexes, functional selectivity, delta opioid receptors, HEK293, multiple conformations
Caractérisation génétique, phénotypique et formation de biofilm des souches de Candida albicans répondant ou non au farnésol

Candida albicans, the most common fungal pathogen, can undergo morphological transitions between yeast and hyphal forms, which are associated with biofilm formation. Farnesol, an endogenous lipid produced by C. albicans, is a quorum sensing molecule that inhibits this transition. Previous work identified two clinical isolates that didn’t respond to farnesol in colony morphology and biofilm assays. Our goal is to better understand C. albicans response to farnesol using these natural farnesol non responders. We have hypothesized that : 1) clinical isolate SC5314, the most characterized strain, is a farnesol responder; 2) non responders’ germination, growth and biofilm formation are different from those of responders; 3) lack of response to farnesol occurs even outside specific culture conditions; 4) farnesol acts through a nuclear receptor that is altered in non responders; 5) difference in farnesol response between strains is explained by transcriptional variations of specific genes (CHK1, HST7, CPH1, GAP1, RAM2 and DPP3), that were previously shown to be potentially involved in farnesol’s mechanism of action.    Non responders produce more hyphae, form 60 % more biofilm and grow 50% slower than responders. The SC5314 strain acts like a responder. Lack of response to farnesol occurs regardless of culture conditions. However, the refractory response to farnesol is not explained by a difference in the proposed genes expression level, except for DPP3 that is upregulated in ATTC® 36802 non responder, suggesting an overproduction of farnesol by this strain. Furthermore, if farnesol acts trough a nuclear receptor, it will be a type not previously described.

Keyphrases: Candida albicans, Biofilm, Quorum sensing, Chk1, Hst7, Cph1, Gap1, Ram2, Dpp3, Farnesol
Interleukin-15 in the pathogenesis of multiple sclerosis and its animal models

Interleukin-15 is a cytokine involved in the homeostatic proliferation and maintenance of CD8 T cells. Activated CD8 T cells are implicated in several autoimmune diseases, including Multiple Sclerosis (MS). Elevated levels of IL-15 have been reported in serum and on peripheral leukocytes of MS patients relative to controls, yet no study has addressed the effects of elevated IL-15 levels on CD8 T cells. To study the in vivo effects of any molecule, the animal model for MS, EAE, is used; the expression of IL-15 during the EAE disease course has not yet been elucidated. Thus the goals of our study were to characterize surface IL-15 expression on human B lymphocytes and determine the effects on human CD8 T cell functions; and to assess the in vivo expression of IL-15 in MS mouse models. We found that B cells are capable of up-regulating the expression of surface IL-15 upon CD40 stimulation, and CD8 T cell effector functions were significantly enhanced upon co-culture with alloreactive IL-15-expressing B cells. In the MS mouse models we used, we found IL-15-expressing immune cells present within the central nervous system (CNS) at various points of disease, and that CNS-infiltrating CD8 T cells were potentially responsive to IL-15. Here, we not only demonstrate the modulation of CD8 T cell responses by IL-15 presented by B cells, implying a role for B cells in MS pathogenesis, but also show the presence of IL-15-expressing cells within the inflamed CNS of EAE.

Keyphrases: Interleukin-15, B cells, CD8 T cells, Multiple Sclerosis, Experimental autoimmune encephalomyelitis
Système de recombinaison Xer chez Staphylococcus aureus

The Xer recombination system is involved in the monomerisation of bacterial replicons, such as plasmids and chromosomes, in a wide variety of bacteria. This system is a site-specific recombination system comprised of two tyrosine recombinases, XerC and XerD, which act in concert to convert dimeric chromosomes to monomers by acting at a specific site near the terminus of replication called the dif site. Xer genes and their site of action have been identified in many gram positive and gram negative bacteria. Staphylococcus aureus represents a gram positive bacterium containing a XerCD/dif system. It is a bacteria implicated in many human diseases, such as skin infections, gastroenteritis and toxic shock syndrome, to name a few. Although the genes encoding the XerC and XerD proteins have been identified, not much is known about their mode of action on the dif site. Mutations in xerC have been obtained, but none in xerD, suggesting that this gene may be essential for this organism. The work presented in this paper has allowed us to better understand the XerD protein of S. aureus, not only in the sequencing of the xerD gene but also in the performing of DNA binding assays. It has been shown that XerD binds to the dif site of E. coli, not only alone but also in cooperativity with E. coli XerC. S. aureus XerD is also capable of complementing the mutated XerD protein in E. coli when it comes to chromosomal recombination. However, it does not demonstrate this same ability to complement XerD regarding recombination at the plasmidic cer sites.

Keyphrases: tyrosine recombinase, XerD, dif, Staphylococcus aureus, Site-specific recombination
Les cellules dendritiques plasmacytoides dans le sang de cordon et après greffe de sang de cordon

Umbilical cord blood transplantation has increasingly been used as a source of hematopoietic stem cells to successfully treat immunodeficiencies and malignant diseases such as leukemia in pediatric patients. Despite important advantages, namely lack of risk for the donor and low incidence of GvHD, use of cord blood is associated with several drawbacks. Specifically, delayed immune reconstitution, more opportunistic infections and a relative risk of relapse are complications that may occur and lead to a poor prognosis. Consequently, new immunotherapeutic strategies should be considered.  In this study, we were interested in plasmacytoid dendritic cells (pDC), whose functions are important for initiation of innate and adaptive immune responses and, in particular, for their ability to activate natural killer cells (NK). In order to elucidate the role and the impact of these cells in cord blood transplantation, pDC and NK numbers and function have been longitudinally followed in cord blood and bone marrow recipients. We showed that pDC and NK cells appeared early after umbilical cord blood transplantation and that these cells retained functional activity. Thus, these cells may constitute a good tool for immunotherapy in umbilical cord blood transplantation.  Moreover, the functional characterization of pDC in cord blood revealed a lower production of IFN-α by cord blood pDC, which may play a role in the lower incidence of GvHD after umbilical cord blood transplantations. In order to determine the molecular mechanism for the negative regulation of IFN-α production by cord blood pDC, we studied the expression of TLR9-IRF7 pathway. The stable expression of TLR9, MyD88, IRAK1 and IRF7 mRNA contrasts with the lower expression of corresponding proteins. Interestingly, expression of microRNA miR-146a and miR-155 is higher in cord blood pDC. Together, these results point to a post-transcriptionnal negative regulation of TLR9-IRF7 pathway which may explain the lower IFN-α production by cord blood pDC. This work reinforces the idea that pDCs constitute a target of choice for developing new therapeutic approaches in cord blood transplantations.

Keyphrases: cord blood, immunotherapie, immunotherapy, hematopoietic stem cell transplantation, plasmacytoid dendritic cells, leukemia
Les infractions portant atteinte à la sécurité du système informatique d’une entreprise

The new information and communication technologies (NICT) currently play an important role in companies, regardless of their size or field of activity; in addition they contribute positively to the economy. However, their use has led to NICT-related criminality, which threatens the security and integrity of the companies’ computer systems. NICT-related criminality has grown exponentially; its increase is hard to assess, and especially hard to control using the existing legislative provisions. Hence, legal adaptations appear unavoidable. Several First World countries have decided to set up, through different means, an adequate legal framework to guarantee the security of companies’ computer systems. Our study will focus precisely on the mechanisms that have been set by two different legal systems. France and Canada, which had to take into account a new reality–new to at least some extent–have decided to amend their respective penal and criminal codes by adding provisions that penalize further infringements. In this work, we will analyze the crimes that undermine the security of the companies’ computer systems in light of the legal tools in place. We will asess how effectively they face today’s computer world and will determine whether or not the law will meet or not the needs of this type of technology.

Keyphrases: New information and communication technologies, Security, Computer system, Company
Embryonic and Uterine Characteristics of Diapause

Delayed implantation or diapause describes arrest or retardation during embryogenesis. In mink, diapause is related to insufficient pituitary prolactin secretion, resulting in incomplete differentiation of the corpus luteum with reduced progesterone concentration. The mink blastocyst at diapause was believed to be totally quiescent or expanding at a low rate. To explore this, DNA replication was studied. Results showed synthesis of DNA, and thus cell proliferation at the morulae stage before diapause and at the blastocyst following activation. DNA replication was detected not only at diapause but also at extended diapause. Furthermore, implantation is considered as a two-way interaction between the blastocyst and the uterus. It has been shown that prostaglandins are important for vascularization of the uterus and products of the prostaglandin pathway could reactivate the mink uterus following diapause. Protein concentration and localization was studied for cytosolic phospholipase A2 (CPLA2) and cyclooxygenase 2 (COX2) in mink uterus.  Expression of CPLA2 and COX2 was up regulated at implantation. It is know that prolactin is the factor that activates the mink corpus luteum. The idea of a link between prolactin and prostaglandin pathway was investigated by quantifying the prolactin receptors in the uterus. Results showed an increase of prolactin receptors at implantation suggesting that prolactin could activate the prostaglandin pathway at the uterus through its own receptor. In conclusion, embryos during diapause are not completely arrested, and proteins related to the prostaglandin pathway are implicated in reactivation of the uterus.

Keyphrases: Diapause, implantation, CPLA2, COX2, PRL-R, DNA replication
Évaluation d’une intervention infirmière Motivationnelle Selon les Stades de Changement (MSSC) sur des comportements d’auto-soins chez des patients insuffisants cardiaques

Heart failure (HF) is characterized by the heart’s reduced capacity to pump, resulting in symptoms causing respiratory problems and reduced quality of life. HF patients are advised to adopt self-care behaviours to prevent frequent episodes of decompensation. While medication and behaviour change play a major role in maintaining physiological stability, patient adherence to self-care recommendations is not optimal. The goal of this pilot study was to evaluate a Motivational Intervention following the Stages of Change (MISC) on HF patients’ self-care behaviours. To guide the MISC, the situation-specific theory of heart failure self-care (Riegel & Dickson, 2008) was retained as well as Bédard et al. (2006) model of intervention, a combination of Motivational interviewing (MI) (Miller & Rollnick, 2006) and Transtheoretical model (TTM) (Prochaska & DiClemente, 1984). This was a randomized experimental pilot project (pre and post-test, with a control group) (n=15/group). Patients in the control group received usual care from the HF clinic while patients from the experimental group (EG) received 3 MISC interventions. Data were collected at one month post-randomization by a blinded research assistant. The effect of the intervention was assessed on five self-care outcomes using analysis of covariance models: self-care maintenance and management (general and specific to HF), confidence in self-care (general and specific to HF) and conviction. Acceptability and feasibility were assessed. Significant results were obtained regarding the confidence in performing self-care behaviours specific to HF. A majority of patient in the EG did progress in the stages of change. The results suggest that a MISC intervention has potential to improve self-care. A larger study is needed to evaluate the effects of the MISC intervention in a randomized controlled trial.

Keyphrases: Self-care, Heart failure, Motivational interviewing, Nursing, Transtheoretical model
Les progressions arithmétiques dans les nombres entiers

The subject of this thesis is the study of arithmetic progressions in the integers. Precisely, we are interested in the size v(N) of the largest subset of the integers from 1 to N that contains no 3 term arithmetic progressions. Therefore, we will construct a large subset of integers with no such progressions, thus giving us a lower bound on v(N). We will begin by looking at the proofs of all the significant lower bounds obtained on v(N), then we will show another proof of the best lower bound known today.  For the proof, we will consider points on a large d-dimensional annulus, and count the number of integer points inside that annulus and the number of arithmetic progressions it contains. To obtain bounds on those quantities, it will be interesting to look at the theory behind counting lattice points in high dimensional spheres, which is the subject of the last section.

Keyphrases: Combinatoire additive, Additive combinatorics, Arithmetic progressions, Lattice theory, Integer points in large spheres
Qualité du sommeil et de la vigilance suite à un ajustement circadien partiel lors d’une simulation de travail de nuit

Night work is associated with many short and long term health problems. Misalignment between the circadian phase and the sleep-wake schedule causes a state of internal desynchrony considered as the main cause of these health problems. Circadian adjustment to night work is proposed as a countermeasure. Usually, complete circadian adjustment is not recommended because night workers experience circadian misalignment when returning on a day-active schedule, causing repeated circadian phase shifts and internal desynchrony. Therefore, partial circadian realignment was proposed as a compromise to stabilize circadian rhythms in night workers. However, the extent of partial circadian adjustment necessary to improve sleep and vigilance quality is still unclear. Here, partial adjustments obtained by phase delay or advance were quantified not only by the phase shift of dim light salivary melatonin onset, but also by the overlap of the melatonin production episode with the sleep-wake cycle during simulated night work. The effects of small but statistically significant partial circadian adjustments on sleep and vigilance quality were assessed in a laboratory simulation of night work to determine their clinical significance. Effects on daytime sleep and night-time vigilance quality were modest. They suggest that even small phase delays may decrease the accumulation of sleep debt, whereas the advance strategy improves subjective night work alertness and mood. Absolute phase shifts, by advance or delay, were associated with improved subjective night work alertness and mood. Field studies are needed, to determine whether these strategies can be adapted to real-life situations and provide effective support to night workers.

Keyphrases: Night Work, Phase Shift, Melatonin, Alertness, Sleep, Vigilance
Influence des facteurs immuno-modulateurs d'origine placentaire sur la différenciation et la maturation des cellules dendritiques plasmacytoïdes

Plasmacytoid dendritic cells (pDC) are specialized dendritic cells, also known as IFN-α producing cells. pDC are key inducers of immune responses following viral infections, by detecting viral antigens through the Toll-like-receptors (TLR) 7 and 9. Although cord blood pDC are unable to produce IFN-α after stimulation, they retain their capacity to mature into antigen presenting cells.   To gain insights into the mechanisms of pDC regulation in cord blood, we investigated the effects of immune regulators secreted by the placenta on the differentiation and function of pDC. Using in vitro differentiated pDC, we analyzed the effect of progesterone (PG), IL-10 and TGF-β, separately or in combination, on pDC differentiation and activation. The data revealed that supra-physiological concentrations of these three factors could individually impair pDC differentiation and IFN-α production. Physiological concentrations of PG, IL-10 or TGF-β, individually, had low impact on pDC while in combination they were able to modulate IFN-α production. We also showed that PG, IL-10 or TGF-β failed to induce the expression of micro-RNA 146a and 155 in pDC. Lastly, our results indicated that PG, IL-10 and TGF-β were present at high levels in cord blood as compared to adult blood.  These results, therefore, shed new light on the synergy of immune regulators secreted by the placenta and their impact on foetal and neonatal immune responses.

Keyphrases: Coord blood, Neonatal immune response, Placenta, Interferon-alpha
Étude sur l’entreposage de la matière grasse chez des femmes obèses post-ménopausées présentant un taux élevé ou normal d’apolipoprotéine B

BACKGROUND: Inefficiency of fatty acid (FA) trapping in white adipose tissue (WAT) is known to promote insulin resistance (IR) in peripheral tissue, but in the liver, it also promotes increased secretion of apolipoprotein B100-lipoproteins (apoB-lipoproteins). We recently demonstrated that post-menopausal obese women with high numbers of fasting apoB-lipoproteins (plasma apoB> 1.2 g/L) had higher IR than women with normal apoB. Our aim was thus to examine whether inefficiency of FA trapping was a mechanism explaining IR in vivo in this population. HYPOTHESES: Postmenopausal overweight and obese women with high apoB have lower efficiency of FA trapping in WAT than women with low apoB. METHODS/RESULTS: The efficiency of FA trapping was examined in 22 non-diabetic overweight and obese women, and the group was separated around median apoB (0.9 g/L) into women with higher vs lower apoB (N=11 per group). The efficiency of FA trapping in WAT was indirectly assessed by following the fate of a 13C-triolein labelled high-fat meal (0.0162g 13C-triolein/g of fat, 66% fat, 47 g fat/m2 body surface area) over 6 hours in circulation (plasma [13C]TG and [13C]FA) and oxidation (breath 13CO2). 13Cenrichment of breath and plasma samples was measured by isotope ratio mass spectrometry. Women with higher apoB had delayed total postprandial plasma TG clearance (p<0.05) but not total plasma free FA clearance compared to women with lower apoB. However, examining the fate of 13C-triolein revealed that women with higher apoB had delayed plasma [13C]-TG clearance (44.78 μM vs 7.81 μM; p<0.05) and plasma [13C]-FA clearance (2.64 μM vs 0.06 μM; p<0.05) at 6 hours, without any differences in % recovered 13C-triolein in breath CO2. This data suggests that women with higher apoB have lower postprandial 13C-triolein clearance and trapping in WAT. CONCLUSION: Ineffective FA trapping in WAT in post-menopausal overweight and obese women with higher apoB may be a mechanism underlying IR in these subjects.

Keyphrases: Apolipoprotein B, Obesity, Insulin resistance, Fatty acid trapping, Dysfunctional adipose tissue, Low Density Lipoprotein (LDL), Postmenopausal women
Le système de recombinaison site-spécifique dif/Xer de Campylobacter jejuni

DNA replication can form dimers in bacteria harboring a circular chromosome. The dif/Xer recombination system resolves monomers them so that chromosome segregation and cell division take place normally. This system is composed of one or two tyrosine recombinases that act at a specific recombination site, dif, with the help of the FtsK translocase that mobilises DNA to the septum before recombination. The Xer system has been first identified and widely characterized in Escherichia coli where XerC and XerD are the recombinases. The system has been found and studied in many other Gram negative and positive bacteria. A different form, carrying a single recombinase acting on an atypical site, has been identified in Streptococci and Lactococci, difSL/XerS.  In silico studies suggested the existence of other single recombinase systems in a sub-group of pathogenic ε-proteobacteriasuch as Campylobacter jejuni and Helicobacter pylori. The components of this system were identified as XerH and difH. In this thesis, the first in vitro studies made on this system are presented. The characterization of the XerH recombinase of C. jejuni started with the sequencing of its gene and with the DNA binding and cleavage assays. These studies showed that XerH could bind difSL of S. suis non-cooperatively, that it could bind difSL half-sites and that it was unable to perform cleavage on difSL. Also, in silico comparisons permitted predictions on FtsK of C. jejuni.

Keyphrases: Tyrosine recombinase, Site-specific recombination, XerH, difH, Campylobacter jejuni