A fluorogenic chemically induced dimerization technology for controlling, imaging and sensing protein proximity.

Molecular tools enabling the control and observation of the proximity of proteins are essential for studying the functional role of physical distance between two proteins. Here we present CATCHFIRE (chemically assisted tethering of chimera by fluorogenic-induced recognition), a chemically induced proximity technology with intrinsic fluorescence imaging and sensing capabilities. CATCHFIRE relies on genetic fusion to small dimerizing domains that interact upon addition of fluorogenic inducers of proximity that fluoresce upon formation of the ternary assembly, allowing real-time monitoring of the chemically induced proximity. CATCHFIRE is rapid and fully reversible and allows the control and tracking of protein localization, protein trafficking, organelle transport and cellular processes, opening new avenues for studying or controlling biological processes with high spatiotemporal resolution. Its fluorogenic nature allows the design of a new class of biosensors for the study of processes such as signal transduction and apoptosis.

Genetic Targeting of Solvatochromic Dyes for Probing Nanoscale Environments of Proteins in Organelles.

A variety of protein tags are available for genetically encoded protein labeling, which allow their precise localization and tracking inside the cells. A new dimension in protein imaging can be offered by combining protein tags with polarity-sensitive fluorescent probes, which provide information about local nanoscale environments of target proteins within the subcellular compartments (organelles). Here, we designed three fluorescent probes based on solvatochromic nile red dye, conjugated to a HaloTag reactive targeting group through polyethylene glycol linkers of varying lengths. The probe with medium linker length, NR12-Halo, was found to label specifically a large variety of proteins localized in defined cell compartments, such as plasma membranes (outer and inner leaflets), endoplasmic reticulum, Golgi apparatus, cytosol, microtubules, actin, and chromatin. Owing to its polarity-sensitive fluorophore, the probe clearly distinguished the proteins localized within apolar lipid membranes from other proteins. Moreover, it revealed dramatic changes in the environment during the life cycle of proteins from biosynthesis to their expected localization and, finally, to recycling inside lysosomes. Heterogeneity in the local polarity of some membrane proteins also suggested a formation of low-polar protein aggregates, for example, within cell-cell contacts. The approach also showed that mechanical stress (cell shrinking by osmotic shock) induced a general polarity decrease in membrane proteins, probably due to the condensation of biomolecules. Finally, the nanoenvironment of some membrane proteins was affected by a polyunsaturated fatty acid diet, which provided the bridge between organization of lipids and proteins. The developed solvatochromic HaloTag probe constitutes a promising tool for probing nanoscale environments of proteins and their interactions within subcellular structures.

PSL Chemical Biology Symposia Third Edition: A Branch of Science in its Explosive Phase.

This symposium is the third PSL (Paris Sciences & Lettres) Chemical Biology meeting (2016, 2019, 2023) held at Institut Curie. This initiative originally started at Institut de Chimie des Substances Naturelles (ICSN) in Gif-sur-Yvette (2013, 2014), under the directorship of Professor Max Malacria, with a strong focus on chemistry. It was then continued at the Institut Curie (2015) covering a larger scope, before becoming the official PSL Chemical Biology meeting. This latest edition was postponed twice for the reasons that we know. This has given us the opportunity to invite additional speakers of great standing. This year, Institut Curie hosted around 300 participants, including 220 on site and over 80 online. The pandemic has had, at least, the virtue of promoting online meetings, which we came to realize is not perfect but has its own merits. In particular, it enables those with restricted time and resources to take part in events and meetings, which can now accommodate unlimited participants. We apologize to all those who could not attend in person this time due to space limitation at Institut Curie.

Fluorescence-Activating and Absorption-Shifting Tags for Advanced Imaging and Biosensing.

Fluorescent labels and biosensors play central roles in biological and medical research. Targeted to specific biomolecules or cells, they allow noninvasive imaging of the machinery that govern cells and organisms in real time. Recently, chemogenetic reporters made of organic dyes specifically anchored to genetic tags have challenged the paradigm of fully genetically encoded fluorescent proteins. Combining the advantage of synthetic fluorophores with the targeting selectivity of genetically encoded tags, these chemogenetic reporters open new exciting prospects for studying cell biochemistry and biology. In this Account, we present the growing toolbox of fluorescence-activating and absorption-shifting tags (FASTs), small monomeric proteins of 14 kDa (125 amino acids residues) that can be used as markers to monitor gene expression and protein localization in live cells and organisms. Engineered by directed protein evolution from the photoactive yellow protein (PYP) from the bacterium Halorhodospira halophila, prototypical FAST binds and stabilizes the fluorescent state of live-cell compatible hydroxybenzylidene rhodanine chromophores. This class of chromophores are normally dark when free in solution or in cells because they dissipate light energy through nonradiative processes. The protein cavity of FAST allows the stabilization of the deprotonated state of the chromophore and blocks the chromophore into a planar conformation, which leads to highly fluorescent protein-chromophore assemblies. The use of such fluorogenic dyes (also called fluorogens) enables the imaging of FAST fusion proteins in cells with high contrast without the need to remove unbound ligands through separate washing steps. Fluorogens with various spectral properties exist nowadays allowing investigators to adjust the spectral properties of FAST to their experimental conditions. Molecular engineering allowed furthermore to generate membrane-impermeant fluorogens for the selective labeling of cell-surface proteins. Over the years, we generated a collection of FAST variants with expanded spectral properties or fluorogen selectivity using a concerted strategy involving molecular engineering and directed protein evolution. Moreover, protein engineering allowed us to adapt FASTs for the design of fluorescent biosensors. Circular permutation enabled the generation of FAST variants with increased conformational flexibility for the design of biosensors in which fluorogen binding is conditioned to the recognition of a given analyte. Bisection of FASTs into two complementary fragments allowed us furthermore to create split variants with reversible complementation that allow the detection and imaging of dynamic protein-protein interactions. We provide, here, a general overview of the current state of development of these different systems and their applications for advanced live cell imaging and biosensing and discuss potential future directions.

Orthogonal fluorescent chemogenetic reporters for multicolor imaging.

Spectrally separated fluorophores allow the observation of multiple targets simultaneously inside living cells, leading to a deeper understanding of the molecular interplay that regulates cell function and fate. Chemogenetic systems combining a tag and a synthetic fluorophore provide certain advantages over fluorescent proteins since there is no requirement for chromophore maturation. Here, we present the engineering of a set of spectrally orthogonal fluorogen-activating tags based on the fluorescence-activating and absorption shifting tag (FAST) that are compatible with two-color, live-cell imaging. The resulting tags, greenFAST and redFAST, demonstrate orthogonality not only in their fluorogen recognition capabilities, but also in their one- and two-photon absorption profiles. This pair of orthogonal tags allowed the creation of a two-color cell cycle sensor capable of detecting very short, early cell cycles in zebrafish development and the development of split complementation systems capable of detecting multiple protein-protein interactions by live-cell fluorescence microscopy.

SARS-CoV-2 testing, infection and places of contamination in France, a national cross-sectional study, December 2021.

BACKGROUND: This study aimed to describe the use of diagnostic testing for SARS-CoV-2 in France until December 2021, the characteristics of people infected, and places of contamination. METHODS: Data were collected from the national 2021 Health Barometer cross-sectional study, which was conducted between February and December 2021 and included French-speaking individuals aged 18-85 years old selected through randomly generated landline and mobile phone numbers. Participants were interviewed about COVID-19-like symptoms in the previous 12 months, diagnostic testing for SARS-CoV-2, positive diagnosis for SARS-CoV-2, and the place(s) of contamination. Determinants of diagnostic testing and of infection were studied using univariate and multivariate Poisson regressions. RESULTS: A total of 24,514 persons participated in the study. We estimated that 66.4% [65.0-67.7] of persons had been tested for SARS-CoV-2 the last time they experienced COVID-19-like symptoms, and that 9.8% [9.3-10.3] of the population in France - with or without symptoms - had been tested positive. Diagnostic testing was less frequent in men, unemployed persons, and people living alone; it was also less frequent during the first months of the pandemic. The estimated proportion of the population infected was higher in healthcare professionals (PRa: 1.5 [1.3-1.7]), those living in large cities ( > = 200 000 inhabitants, and Paris area) (1.4 [1.2-1.6]), and in households comprising > 3 persons (1.7 [1.5-2.0]). It was lower in retired persons (0.8 [0.6-0.97]) and those over 65 years old (0.6 [0.4-0.9]). Almost two-thirds (65.7%) of infected persons declared they knew where they were contaminated; 5.8% [4.5-7.4] reported being contaminated outdoors, 47.9% [44.8-51.0] in unventilated indoor environments, and 43.4% [40.3-46.6] in ventilated indoor environments. Specifically, 51.1% [48.0-54.2] declared they were contaminated at home or in a family of friend's house, 29.1% [26.4-31.9] at their workplace, 13.9% [11.9-16.1] in a healthcare structure, and 9.0% [7.4-10.8] in a public eating place (e.g., cafeteria, bar, restaurant). CONCLUSIONS: To limit viral spread, preventive actions should preferentially target persons tested least frequently and those at a higher risk of infection. They should also target contamination in households, healthcare structures, and public eating places. Importantly, contamination is most frequent in places where prevention measures are most difficult to implement.

Fluorescent secreted bacterial effectors reveal active intravacuolar proliferation of Listeria monocytogenes in epithelial cells.

Real-time imaging of bacterial virulence factor dynamics is hampered by the limited number of fluorescent tools suitable for tagging secreted effectors. Here, we demonstrated that the fluorogenic reporter FAST could be used to tag secreted proteins, and we implemented it to monitor infection dynamics in epithelial cells exposed to the human pathogen Listeria monocytogenes (Lm). By tracking individual FAST-labelled vacuoles after Lm internalisation into cells, we unveiled the heterogeneity of residence time inside entry vacuoles. Although half of the bacterial population escaped within 13 minutes after entry, 12% of bacteria remained entrapped over an hour inside long term vacuoles, and sometimes much longer, regardless of the secretion of the pore-forming toxin listeriolysin O (LLO). We imaged LLO-FAST in these long-term vacuoles, and showed that LLO enabled Lm to proliferate inside these compartments, reminiscent of what had been previously observed for Spacious Listeria-containing phagosomes (SLAPs). Unexpectedly, inside epithelial SLAP-like vacuoles (eSLAPs), Lm proliferated as fast as in the host cytosol. eSLAPs thus constitute an alternative replication niche in epithelial cells that might promote the colonization of host tissues.

Arylopeptoid oligomers functionalised by combinatorial or sequential on-resin click chemistry using a copper(I)-N-heterocyclic carbene catalyst.

An efficient on-resin click chemistry protocol using a stable copper(I)-N-heterocyclic carbene catalyst is developed for post-functionalization of N-alkylated aminomethylbenzamide oligomers (arylopeptoids). The accessibility to a panel of polyfunctionalized N-substituted aromatic oligoamides by solid-phase synthesis is demonstrated using combinatorial and sequential approaches.

An expanded palette of fluorogenic HaloTag probes with enhanced contrast for targeted cellular imaging.

We report the development of HaloTag fluorogens based on dipolar flexible molecular rotor structures. By modulating the electron donating and withdrawing groups, we have tuned the absorption and emission wavelengths to design a palette of fluorogens with emissions spanning the green to red range, opening new possibilities for multicolor imaging. The probes were studied in glycerol and in the presence of HaloTag and exhibited good fluorogenic properties thanks to a viscosity-sensitive emission. In live-cell confocal imaging, the fluorogens yielded only a very low non-specific signal that enabled wash-free targeted imaging of intracellular organelles and proteins with good contrast. Combining experimental studies and theoretical investigation of the protein/fluorogen complexes by molecular dynamics, these results offer new insight into the design of molecular rotor-based fluorogenic HaloTag probes in order to improve reaction rates and the imaging contrast.

Influenza vaccination coverage of professionals working in nursing homes in France and related determinants, 2018-2019 season: a cross-sectional survey.

BACKGROUND: The burden of influenza morbidity and mortality in nursing homes (NH) is high. Vaccination of residents and professionals working in NH is the main prevention strategy. Despite recommendations, vaccination coverage among professionals is generally low. METHODS: We performed a nationwide cross-sectional survey of NH using a single-stage stratified random sampling design to estimate influenza vaccination coverage in NH healthcare workers (HCW) and non-medical professionals in France during the 2018-2019 season, and to identify measures likely to increase it. For each NH, a questionnaire was completed with aggregated data by one member of the management team. A multivariate analysis was performed using a negative binomial regression. RESULTS: Five-hundred and eighty nine NH filled in the study questionnaire (response rate: 49.5%). When considering all professionals (i.e., HCW and non-medical professionals), overall vaccination coverage was 30.6% (95%CI [28.2-33.0], range: 1.6-96.2). Overall influenza vaccination coverage in HCW was 31.9% [29.7-34.1]. It varied according to occupational category: 75.5% [69.3-81.7] for physicians, 42.9% [39.4-46.4] for nurses, 26.7% [24.5-29.0] for nursing assistants, and 34.0% [30.1-38.0] for other paramedical personnel. Vaccination coverage was higher i) in private nursing homes (RRa: 1.3, [1.1-1.5]), ii) in small nursing homes (0.9 [0.8-0.9]), iii) when vaccination was offered free of charge (1.4, [1.1-1.8]), iv) when vaccination promotion for professionals included individual (1.6 [1.1-2.1]) or collective (1.3 [1.1-1.5]) information sessions, videos or games (1.4 [1.2-1.6], v) when information on influenza vaccines was provided (1.2 [1.0-1.3], and finally, vi) when a vaccination point of contact-defined as an HCW who could provide reliable information on vaccination-was nominated within the nursing home (1.7 [1.3-2.2]). CONCLUSIONS: Urgent and innovative actions are required to increase coverage in HCW. Vaccination programmes should include free on-site vaccination and education campaigns, and particularly target nursing assistants. The results of this nationwide study provide keys for improving influenza vaccination coverage in HCW. Programmes should ensure that information on influenza vaccines is provided by a vaccination point of contact in NH using attractive media. Combining the different prevention measures proposed could increase coverage in NH nationwide by over 50%.

Increased awareness and knowledge of Lyme Borreliosis and tick bite prevention among the general population in France: 2016 and 2019 health barometer survey.

BACKGROUND: Lyme borreliosis (LB) is the most frequent tick-borne disease in France. In the absence of a vaccine, LB prevention mainly relies on reducing tick bites. In 2016, the French Ministry of Health launched a national plan against tick-borne infections, including a prevention component. To evaluate the impact of this prevention strategy, we assessed knowledge and practices of tick bite prevention using the 2016 and 2019 national surveys on health attitudes and beliefs known as the French Health Barometer. METHODS: The Health Barometer is a repeated nationwide phone survey conducted annually on a random sample aged 18 to 75 years living in mainland France. In 2016 and 2019, participants were asked, among others, about their exposure to ticks, their behavior and practices regarding tick bites, and their knowledge about LB and its prevention. RESULTS: In 2019, 30% of the population reported a lifetime tick bite and 6% in the last year, an increase from 25% and 4%, respectively, in 2016 (p < 0.001). In 2019, 25% of the population felt exposed to tick bites compared to 23% in 2016 (p < 0.001). The proportion of participants who had heard about LB and who considered themselves well informed respectively increased from 66% and 29% in 2016 to 79% and 41% in 2019, (p < 0.001). In 2019 compared to 2016, a greater part of the French population applied protective measures against tick bites, particularly wearing protective clothing (74% vs 66%, p < 0.001) and regular tick checks and prompt tick removal after exposure (54% vs 47%, p < 0.001). CONCLUSIONS: A substantial proportion of French residents are exposed to tick bites and apply protective measures. Our findings indicate a trend toward an increased knowledge and awareness of tick bites and LB between 2016 and 2019 in France. Our results can be used to target future information campaigns to specific age groups or at-risk areas in addition to the general population. However, we need to further study the barriers to the use of preventive measures.

Fluorogenic Protein-Based Strategies for Detection, Actuation, and Sensing.

Fluorescence imaging has become an indispensable tool in cell and molecular biology. GFP-like fluorescent proteins have revolutionized fluorescence microscopy, giving experimenters exquisite control over the localization and specificity of tagged constructs. However, these systems present certain drawbacks and as such, alternative systems based on a fluorogenic interaction between a chromophore and a protein have been developed. While these systems are initially designed as fluorescent labels, they also present new opportunities for the development of novel labeling and detection strategies. This review focuses on new labeling protocols, actuation methods, and biosensors based on fluorogenic protein systems.

A Far-Red Emitting Fluorescent Chemogenetic Reporter for In†Vivo Molecular Imaging.

Far-red emitting fluorescent labels are highly desirable for spectral multiplexing and deep tissue imaging. Here, we describe the generation of frFAST (far-red Fluorescence Activating and absorption Shifting Tag), a 14-kDa monomeric protein that forms a bright far-red fluorescent assembly with (4-hydroxy-3-methoxy-phenyl)allylidene rhodanine (HPAR-3OM). As HPAR-3OM is essentially non-fluorescent in solution and in cells, frFAST can be imaged with high contrast in presence of free HPAR-3OM, which allowed the rapid and efficient imaging of frFAST fusions in live cells, zebrafish embryo/larvae, and chicken embryos. Beyond enabling the genetic encoding of far-red fluorescence, frFAST allowed the design of a far-red chemogenetic reporter of protein-protein interactions, demonstrating its great potential for the design of innovative far-red emitting biosensors.

2nd PSL Chemical Biology Symposium (2019): At the Crossroads of Chemistry and Biology.

Chemical Biology is the science of designing chemical tools to dissect and manipulate biology at different scales. It provides the fertile ground from which to address important problems of our society, such as human health and environment.

HCV and HBV prevalence based on home blood self-sampling and screening history in the general population in 2016: contribution to the new French screening strategy.

BACKGROUND: The advent of effective direct-acting antivirals (DAAs), has prompted an assessment of the French Hepatitis C virus (HCV) screening strategy, which historically targeted high-risk groups. One of the options put forward is the implementation of combined (i.e., simultaneous) HCV, Hepatitis B virus (HBV) and HIV screening for all adults at least once during their lifetime ("universal combined screening"). However, recent national survey-based data are lacking to guide decision-making regarding which new strategy to implement. Accordingly, we aimed to provide updated data for both chronic hepatitis C (CHC) and B (CHB) prevalence and for HCV and HBV screening history, using data from the BaroTest and 2016 Health Barometer (2016-HB) studies, respectively. METHODS: 2016-HB was a national cross-sectional phone based health survey conducted in 2016 among 20,032 randomly selected individuals from the general population in mainland France. BaroTest was a virological sub-study nested in 2016-HB. Data collected for BaroTest were based on home blood self-sampling on dried blood spots (DBS). RESULTS: From 6945 analyzed DBS, chronic hepatitis C (CHC) and B (CHB) prevalence was estimated at 0.30% (95% Confidence Interval (CI): 0.13-0.70) and 0.30% (95% CI: 0.13-0.70), respectively. The proportion of individuals aware of their status was estimated at 80.6% (95% CI: 44.2-95.6) for CHC and 17.5% (95% CI: 4.9-46.4) for CHB. Universal combined screening would involve testing between 32.6 and 85.3% of 15-75 year olds according to whether we consider only individuals not previously tested for any of the three viruses, or also those already tested for one or two of the viruses. CONCLUSIONS: Our data are essential to guide decision-making regarding which new HCV screening recommendation to implement in France. They also highlight that efforts are still needed to achieve the WHO's targets for eliminating these diseases. Home blood self-sampling may prove to be a useful tool for screening and epidemiological studies.

Using hydrogenated and perfluorinated gases to probe the interactions and structure of fluorinated ionic liquids.

After studying the properties of a mixture of hydrogenated and fluorinated ionic liquids we have measured the solubility of perfluoromethane, perfluoroethane and perfluoropropane in 1-alkyl-3-methylimidazolium based ionic liquids with hydrogenated or fluorinated alkyl side-chains: 1-octyl-3-methylimidazolium bis[trifluoromethylsulfonyl]amide ([C8C1Im][NTf2]), 1-octyl-3-methylimidazolium bis[pentafluoroethylsulfonyl]amide ([C8C1Im][BETI]), 1-(3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl)-3-methylimidazolium bis[trifluoromethylsulfonyl]amide ([C8H4F13C1Im][NTf2]), and 1-(3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl)-3-methylimidazolium bis[pentafluoroethylsulfonyl]amide ([C8H4F13C1Im][BETI]). The ionic liquids expand on mixing and mix endothermally with a relatively high enthalpy of mixing (ΔmixH for [C8C1Im]x[C8H4F13C1Im](1-x)[NTf2] of ca. 0.85 kJ mol-1 for x = 0.5) when compared with other ionic mixtures. The solubility of the perfluorinated gases is larger in the fluorinated ionic liquids when compared with that of their hydrogenated counterparts and follows the order [C8H4F13C1Im][BETI] > [C8H4F13C1Im][NTf2] > [C8C1Im][BETI] > [C8C1Im][NTf2], a behaviour explained by a slightly more favourable enthalpy of solvation. The fluorinated ionic liquids nevertheless do not dissolve larger quantities of perfluorinated gases than their hydrogenated equivalents, as observed by comparing the results herein for perfluoroethane to those measured previously for ethane in the same ionic liquids. By using molecular simulations to study the microscopic structure of the solutions, we could show that the gases, hydrogenated and fluorinated, are always preferentially solvated in the apolar domains of the ionic liquids, and the hydrogenated hydrocarbon gases are always more soluble, independent of the fluorination of the ionic liquid.

General practitioners who never perform Pap smear: the medical offer and the socio-economic context around their office could limit their involvement in cervical cancer screening.

BACKGROUND: In France, with the growing scarcity of gynecologists and a globally low and socially differentiated coverage of cervical cancer screening (CCS), general practitioners (GPs) are valuable resources to improve screening services for women. Still all GPs do not perform Pap smears. In order to promote this screening among GPs, the characteristics of physicians who never perform CCS should be more precisely specified. Besides already-known individual characteristics, the contextual aspects of the physicians' office, such as gynecologist density in the area, could shape GPs gynecological activities. METHODS: To analyze county (département) characteristics of GPs' office associated with no performance of CCS, we used a representative sample of 1063 French GPs conducted in 2009 and we constructed mixed models with two levels, GP and county. RESULTS: Almost 35% (n = 369) of the GPs declared never performing CCS. GPs working in counties with a poor GP-density per inhabitants were more likely to perform CCS (odds ratio (OR) = 0.52 for each increase of density by 1 GP per 10,000 inhabitants, 95% confidence interval (CI) = 0.37-0.74). On the contrary, GPs working in counties with an easier access to a gynecologist were more likely not to perform CCS (OR = 1.06 for each increase of density by 1 gynecologist per 100,000 women, 95%CI = 1.03-1.10 and OR = 2.02 if the first gynecologist is reachable in less than 15 min, 95%CI = 1.20-3.41) as well as GPs working in areas with a poverty rate above the national average (OR = 1.66, 95%CI = 1.09-2.54). These contextual characteristics explain most of the differences between counties concerning rates of not performing CCS. CONCLUSIONS: Specific programs should be developed for GPs working in contexts unfavorable to their involvement in CCS.

Small fluorescence-activating and absorption-shifting tag for tunable protein imaging in vivo.

This paper presents Yellow Fluorescence-Activating and absorption-Shifting Tag (Y-FAST), a small monomeric protein tag, half as large as the green fluorescent protein, enabling fluorescent labeling of proteins in a reversible and specific manner through the reversible binding and activation of a cell-permeant and nontoxic fluorogenic ligand (a so-called fluorogen). A unique fluorogen activation mechanism based on two spectroscopic changes, increase of fluorescence quantum yield and absorption red shift, provides high labeling selectivity. Y-FAST was engineered from the 14-kDa photoactive yellow protein by directed evolution using yeast display and fluorescence-activated cell sorting. Y-FAST is as bright as common fluorescent proteins, exhibits good photostability, and allows the efficient labeling of proteins in various organelles and hosts. Upon fluorogen binding, fluorescence appears instantaneously, allowing monitoring of rapid processes in near real time. Y-FAST distinguishes itself from other tagging systems because the fluorogen binding is highly dynamic and fully reversible, which enables rapid labeling and unlabeling of proteins by addition and withdrawal of the fluorogen, opening new exciting prospects for the development of multiplexing imaging protocols based on sequential labeling.

Population perception of mandatory childhood vaccination programme before its implementation, France, 2017.

BackgroundVaccination policy in France was previously characterised by the coexistence of eight recommended and three mandatory vaccinations for children younger than 2 years old. These 11 vaccines are now mandatory for all children born after 1 January 2018.AimTo study the French population's opinion about this new policy and to assess factors associated with a positive opinion during this changing phase.MethodsA cross-sectional survey about vaccination was conducted from 16 November-19 December 2017 among the GrippeNet.fr cohort. Data were weighted for age, sex and education according to the French population. Univariate and multivariate analyses were performed to identify factors associated with a favourable opinion on mandatory vaccines' extension and defined in the '3Cs' model by the World Health Organization Strategic Advisory Group of Experts working group on vaccine hesitancy.ResultsOf the 3,222 participants (response rate 50.5%) and after adjustment, 64.5% agreed with the extension of mandatory vaccines. It was considered a necessary step by 68.7% of the study population, while 33.8% considered it unsafe for children and 56.9% saw it as authoritarian. Factors associated with a positive opinion about the extension of mandatory vaccines were components of the confidence, complacency and convenience dimensions of the '3Cs' model.ConclusionsIn our sample, two thirds of the French population was in favour of the extension of mandatory vaccines for children. Perception of vaccine safety and benefits were major predictors for positive and negative opinions about this new policy.

Next-Generation Fluorogen-Based Reporters and Biosensors for Advanced Bioimaging.

Our ability to observe biochemical events with high spatial and temporal resolution is essential for understanding the functioning of living systems. Intrinsically fluorescent proteins such as the green fluorescent protein (GFP) have revolutionized the way biologists study cells and organisms. The fluorescence toolbox has been recently extended with new fluorescent reporters composed of a genetically encoded tag that binds endogenously present or exogenously applied fluorogenic chromophores (so-called fluorogens) and activates their fluorescence. This review presents the toolbox of fluorogen-based reporters and biosensors available to biologists. Various applications are detailed to illustrate the possible uses and opportunities offered by this new generation of fluorescent probes and sensors for advanced bioimaging.