The French version of the HSCL-25 has now been validated for use in primary care

Background The Hopkins Symptom Checklist in 25 items (HSCL-25) helps to assess anxiety and depression in Primary Care. Anxiety and depression show considerable overlap in primary care. This self-administrated questionnaire is valid, reliable and ergonomic in the original US version. We have translated it into French. The aim of this study was to estimate the test characteristics of the HSCL-25, in its French version (F-HSCL-25), by comparing it to the Present State Examination-9 French version (F-PSE-9) and by determining its internal validity and dimensions. Method Outpatients from three French General Practice settings (rural, semi-rural and urban) were recruited: approximately 20,000 outpatients among 17 GPs. Two groups were formed: F-HSCL-25 ≥1.75 and F-HSCL-25 1.75 was considered to indicate a clinically relevant level of symptoms of depression and anxiety. In order to obtain two balanced groups, a different method of randomization was chosen for each group. The F-PSE-9 was randomly administered to 1 in 2 patients in the F-HSCL-25 ≥1.75 group, and to 1 in 16 in the (much larger) F-HSCL-25 <1.75 group. The diagnostic performance was assessed and the test results obtained from both groups were compared with their F-PSE-9 results. Results Of the 1126 patients who completed the F-HCL-25, 886 joined the F-HSCL-25 <1.75 group and 240 the F-HSCL-25 ≥1.75 group. The overall prevalence of depression, using the F-HSCL-25, was 21% in these medical practices. The diagnostic performance of the F-HSCL-25 versus the F-PSE-9, the external criteria were as follows: Positive Predictive Value (PPV) 69.8%, Negative Predictive Value (NPV) 87%; Sensitivity 59.1%, and Specificity 91.4%. The Principal Component Analysis showed that F-HSCL-25 is a one-dimensional tool (anxiety and depression dimensions combined) with a Cronbach Alpha of 0.93. Conclusion The F-HSCL-25 is an appropriate diagnostic tool for anxiety and depression in primary care in France due to its high specificity and high NPV. The HSCL-25 scale has a high eigenvalue. This pilot study will be extended throughout Europe; however, preliminary evidence suggests that the HSCL-25 is a reliable and suitable diagnostic tool for primary care

Arsonium-Containing Lipophosphoramides, Poly-Functional Nano-Carriers for Simultaneous Antibacterial Action and Eukaryotic Cell Transfection.

International audienceGene therapy of diseases like cystic fibrosis (CF) would consist of delivering a gene medicine towards the lungs via the respiratory tract into the target epithelial cells. Accordingly, poly-functional nano-carriers are required in order to overcome the various successive barriers of such a complex environment, such as airway colonization with bacterial strains. In this work, the antibacterial effectiveness of a series of cationic lipids is investigated before evaluating its compatibility with gene transfer into human bronchial epithelial cells. Among the various compounds considered, some bearing a trimethyl-arsonium headgroup demonstrate very potent biocide effects towards clinically relevant bacterial strains. In contrast to cationic lipids exhibiting no or insufficient antibacterial potency, arsonium-containing lipophosphoramides can simultaneously inhibit bacteria while delivering DNA into eukaryotic cells, as efficiently and safely as in absence of bacteria. Moreover, such vectors can demonstrate antibacterial activity in vitro while retaining high gene transfection efficiency to the nasal epithelium as well as to the lungs in mice in vivo. Arsonium-containing amphiphiles are the first synthetic compounds shown to achieve efficient gene delivery in the presence of bacteria, a property particularly suitable for gene therapy strategies under infected conditions such as within the airways of CF patients

Interaction between galectin-3 and cystinosin uncovers a pathogenic role of inflammation in&nbsp;kidney involvement of cystinosis.

Inflammation is involved in the pathogenesis of many disorders. However, the underlying mechanisms are often unknown. Here, we test whether cystinosin, the protein involved in cystinosis, is a critical regulator of galectin-3, a member of the β-galactosidase binding protein family, during inflammation. Cystinosis is a lysosomal storage disorder and, despite ubiquitous expression of cystinosin, the kidney is the primary organ impacted by the disease. Cystinosin was found to enhance lysosomal localization and degradation of galectin-3. In Ctns-/- mice, a mouse model of cystinosis, galectin-3 is overexpressed in the kidney. The absence of galectin-3 in cystinotic mice ameliorates pathologic renal function and structure and decreases macrophage/monocyte infiltration in the kidney of the Ctns-/-Gal3-/- mice compared to Ctns-/- mice. These data strongly suggest that galectin-3 mediates inflammation involved in kidney disease progression in cystinosis. Furthermore, galectin-3 was found to interact with the pro-inflammatory cytokine Monocyte Chemoattractant Protein-1, which stimulates the recruitment of monocytes/macrophages, and proved to be significantly increased in the serum of Ctns-/- mice and also patients with cystinosis. Thus, our findings highlight a new role for cystinosin and galectin-3 interaction in inflammation and provide an additional mechanistic explanation for the kidney disease of cystinosis. This may lead to the identification of new drug targets to delay cystinosis progression

Nine forward–backward translations of the Hopkins symptom checklist-25 with cultural checks

Introduction: The Hopkins Symptom Checklist-25 (HSCL-25) is an effective, reliable, and ergonomic tool that can be used for depression diagnosis and monitoring in daily practice. To allow its broad use by family practice physicians (FPs), it was translated from English into nine European languages (Greek, Polish, Bulgarian, Croatian, Catalan, Galician, Spanish, Italian, and French) and the translation homogeneity was confirmed. This study describes this process. Methods: First, two translators (an academic translator and an FP researcher) were recruited for the forward translation (FT). A panel of English-speaking FPs that included at least 15 experts (researchers, teachers, and practitioners) was organized in each country to finalize the FT using a Delphi procedure. Results: One or two Delphi procedure rounds were sufficient for each translation. Then, a different translator, who did not know the original version of the HSCL-25, performed a backward translation in English. An expert panel of linguists compared the two English versions. Differences were listed and a multicultural consensus group determined whether they were due to linguistic problems or to cultural differences. All versions underwent cultural check. Conclusion: All nine translations were finalized without altering the original meaning

Immunothérapie antitumorale (production de lymphocytes T spécifiques d antigènes de tumeur au moyen de cellules présentatrices d antigènes génétiquement modifiées)

L'immunothérapie par stimulation des lymphocytes T spécifiques d'antigènes de tumeur constitue une approche très prometteuse pour le traitement des cancers. Notre travail a porté sur la vectorisation de l'antigène de tumeur NY-ESO-1 dans les DC, à l'aide d'un vecteur synthétique du groupe des lipophosphoramides cationiques, le KLN-5. Nous avons dans un premier temps déterminé les conditions optimales de transfection. Nous avons montré que la protéine NY-ESO-1 est synthétisée dans les DC transfectées, clivée en peptides puis chargée sur les molécules de classe I et finalement présentée aux lymphocytes T de façon efficace. L'activation d'un clone spécifique de l'épitope de classe II a aussi été obtenue, mais reste à confirmer. Dans le seconde partie de notre travail, nous avons utilisé des cellules présentatrices d'antigène artificielles (CPAA), modifiées pour exprimer de façon stable un complexe HLA-peptide, ainsi que les molécules humaines de costimulation B7.1, ICAM-1, et LFA-3, afin de stimuler in vitro des lymphocytes T cytotoxiques, spécifiques de l'antigène NY-ESO-1. Nous avons pu détecter la présence de lymphocytes T CD8 spécifiques dans la population stimulée par les CPAA exprimant le peptide NY-ESO-1 par marquage tétramère. En conclusion, ce travail nous a permis d'évaluer deux méthodes en vue d'activer des lymphocytes T in vitro. L'utilisation de vecteurs synthétiques pour modifier génétiquement les DC semble être une voie prometteuse pour générer une réponse polyclonale T CD8 et T CD4 contre des antigènes de tumeurs. Enfin, les CPAA représentent un système alternatif de production de lymphocytes T spécifique d'antigènes de tumeurs à visée d'immunothérapie adoptive.Genetic modification of human monocyte-derived dendritic cells (DCs) with tumor-associated antigen (TAA) cDNA sequences is a promising strategy for immunotherapy of cancer. As an alternative to the commonly used viral vectors, a nonviral gene transfer method based on the use of the lipophosphoramide transfection reagent KLN-5 was investigated. We first determined the optimum conditions for transfection. The NY-ESO-1 TAA coding gene was found efficiently transfected in DCs as shown by tumor antigen mRNA expression analysis. Antigen processing and presentation of the immunodominant epitope in the HLA-A2 context was demonstrated by specific activation of a NY-ESO-1 CD8+ T cell clone. A NY-ESO-1 specific CD4+ T clone activation was also observed but remains to be confirmed. In the second part of our work, artificial APC (AAPC) were used to stimulate NY-ESO-1 specific cytotoxic T cells (CTL). The CPAA are derived from mouse fibroblasts, retrovirally transduced to stably express a HLA-peptide complex, and the B7.1, ICAM-1, and LFA-3. Stimulation experiments were performed on naive T cells. NY-ESO-1 specific CD8 T cells were detected by tetramer staining when stimulated with the AAPC expressing NY-ESO-1. But we were not able to detect INF-g production when CTL were stimulated with NY-ESO-1 expressing cells. In conclusion, in this work we have evaluated two T cell activation methods. The synthetic vector KLN5 represents an attractive nonviral agent to generate a T CD8 and T CD4 response against TAA, even if the ability to prime a naïve T cell response remains to be shown. And finally, AAPC are useful to study T cell activation and to induce antigen-specific T cells for clinical purposes.RENNES1-BU Sciences Philo (352382102) / SudocSudocFranceF

Editorial: Current status and prospects on nucleic acid transfer

International audienc

Thiophosphoramidates : une nouvelle famille de lipides pour la vectorisation d’acides nucléiques

Patent PCT/EP2011/06742

Aerosol Delivery to the Airways Using Cationic Lipid Nanocomplexes in a Perspective of Cystic Fibrosis Treatment

International audienceDiseases affecting the respiratory tract are numerous and many are disabling or deadly. Nucleic acids are emerging as a new tool and a new therapeutic class of compounds that could be used to address some lung diseases. However, considering the different anatomical barriers and the physicochemical behavior of nucleic acids in the blood stream, lungs are not easy to treat following a general administration. In this context, aerosol delivery is largely considered as an adapted and noninvasive approach avoiding the hepatic metabolism. This local delivery leads to a lung concentration and limits eventual side effects associated with a systemic injection. However, aerosol administration of active molecules is a real challenge due to the physical, chemical, and biological obstacles, such as respiratory movement, surfactant, or bacterial strains. Nanocomplexes have the potential to increase bioavailability and favor intracellular penetration of specific drugs into the pulmonary tissue. In this chapter, we will present our strategy to develop efficient gene carriers that, from a synthesis point of view, can be prepared on a large scale. We will discuss their use in preparing various formulations and their progressive adaptation to the airway constraints. In particular, we will focus our attention on cationic amphiphilic complexes used for aerosol gene delivery and we will present promising formulations in preclinical studies in a cystic fibrosis context.Les maladies touchant les voies respiratoires sont nombreuses et parfois mortelles. Les acides nucléiques apparaissent comme de nouveaux outils et une nouvelle classe thérapeutique qui pourraient être utilisés vis-à-vis de certaines maladies pulmonaires. Cependant, considérant les différentes barrières anatomiques et les caractéristiques physico-chimiques des acides nucléiques placés dans le flux sanguin, les poumons ne sont pas facile à traiter par une approche systémique. Dans ce contexte, l’aérosolisation, qui est une administration non invasive, est une approche pertinente. Ainsi, cette délivrance locale peut aussi contribuer à éviter des effet secondaires associés à une délivrance systémique. Néanmoins, la délivrance par aérosol est un véritable défi du fait des obstacles physiques, chimiques et biologiques tel que le mouvement respiratoire, la présence de surfactants ou de bactéries. Les nano-complexes ont le potentiel d’augmenter la biodisponibilité et de favoriser la pénétration intracellulaire. Dans ce chapitre, nous présentons nos stratégies pour développer des transporteurs de gènes efficaces. Cela inclut la conception des nano-vecteurs et leurs évaluations. Un focus est fait sur les amphiphiles cationiques qui sont utilisés pour une administration de plasmide par aérosols. Nous montrons que certaines formulations produisent des résultats précliniques intéressants et cela dans le contexte de la mucoviscidose

Phosphonodithioester–Amine Coupling as a Key Reaction Step for the Design of Cationic Amphiphiles Used for Gene Delivery

International audienceA convergent synthesis of cationic amphiphilic compounds is reported here with the use of the phosphonodithioester–amine coupling (PAC) reaction. This versatile reaction occurs at room temperature without any catalyst, allowing binding of the lipid moiety to a polar head group. This strategy is illustrated with the use of two lipid units featuring either two oleyl chains or two-branched saturated lipid chains. The final cationic amphiphiles were evaluated as carriers for plasmid DNA delivery in four cell lines (A549, Calu3, CFBE and 16HBE) and were compared to standards (BSV36 and KLN47). These new amphiphilic derivatives, which were formulated with DOPE or DOPE-cholesterol as helper lipids, feature high transfection efficacies when associated with DOPE. The highest transfection efficacies were observed in the four cell lines at low charge ratios (CR = 0.7, 1 or 2). At these CRs, no toxic effects were detected. Altogether, this new synthesis scheme using the PAC reaction opens up new possibilities for investigating the effects of lipid or polar head groups on transfection efficacies

Cutevariant: a standalone GUI-based desktop application to explore genetic variations from an annotated VCF file

International audienceAbstract Summary Cutevariant is a graphical user interface (GUI)-based desktop application designed to filter variations from annotated VCF file. The application imports data into a local SQLite database where complex filter queries can be built either from GUI controllers or using a domain-specific language called Variant Query Language. Cutevariant provides more features than existing applications and is fully customizable thanks to a complete plugins architecture. Availability and implementation Cutevariant is distributed as a multiplatform client-side software under an open source license and is available at https://github.com/labsquare/cutevariant