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The taxonomic status of the keeled back mullets (Teleostei: Mugilidae) has been reinvestigated. Two nominal mugilid species having keeled backs from East Asia: Mugil lauvergnii Eydoux & Souleyet, 1850 and Mugil affinis Günther, 1861 have been re-evaluated through examination of the holotypes and fresh specimens. Comparison of morpho-meristic characters of the holotypes shows that both species are identical. Phylogenetic analysis based on mitochondrial cytochrome c oxidase 1 (CO1) confirmed morphological data by highlighting presence of a single clade from East Asia. Mugil lauvergnii (=Planiliza lauvergnii) is thus the sole keeled back mullet from East Asia and a senior synonym of Mugil affinis (=Planiliza affinis). The taxonomic status of two other keeled back mullets, Planiliza carinata and P. klunzingeri, is also contentious due to their similar morphology. Meristic and morphometric variation as well as sequence divergence between the two species are limited but phylogenetic analyses delineate well-supported clades consistent with biogeography and currently accepted taxonomy. Planiliza carinata and P. klunzingeri share a recent common ancestor in a Maximum Likelihood tree, with separate distribution ranges while P. lauvergnii formed a paraphyletic lineage. Based on present findings, we suggest maintenance of the taxonomic distinction of P. klunzingeri and P. carinata and discuss its evolutionary significance.

New data based on multigene phylogenetic analyses using the COI, 16S, and cytb genes and subsequent molecular species delimitation revealed that the mugilid genus Chelon contains 11 species. Of these, two species, Chelon sp. A and Chelon sp. B, remained unidentified in previous studies. While Chelon sp. B seemingly is a close relative to C. dumerili (Steindachner, 1870), no more detailed information were provided for Chelon sp. A which is present along the east coast of South Africa. Genetic analyses performed in this study, revealed that specimens from the Arabian/Persian Gulf and the Red Sea are nested within one genetic lineage alongside Chelon sp. A. Morphological, morphometric, and meristic analyses of specimens from all three areas demonstrated that they belong to Chelon persicus Senou, Randall & Okyiama, 1995. Chelon persicus was originally described in the genus Chelon but was later placed either in the genus Liza or Planiliza. However, results presented herein confirm its placement in Chelon and its distribution range is extended in the Western Indian Ocean from the type locality (Arabian/Persian Gulf) west to the Red Sea and south to South Africa. Further, the uncertain validity of the recently described Chelon caeruleus Deef, 2018 from the Mediterranean Sea and a previously proposed generic character distinguishing Chelon from Planiliza and Parachelon, the shape of the paired postzygapophyses of the second vertebra, are briefly discussed.

Bacterial AB toxins are secreted virulence factors that are internalized by target cells through receptor-mediated endocytosis, translocating their enzymatic domain to the cytosol from endosomes (short-trip) or the endoplasmic reticulum (long-trip). To accomplish this, they evolved a multidomain structure organized into either a single polypeptide chain or non-covalently associated polypeptide chains. The prototypical short-trip single-chain toxin is characterized by a receptor-binding domain that confers cellular specificity and a translocation domain responsible for pore formation whereby the catalytic domain translocates to the cytosol in an endosomal acidification-dependent way.In this work, the resolution of the three-dimensional structure of AIP56 showed that instead of a two-domain organization, as previously predicted, AIP56 has a three-domain organization, with a NleC-like catalytic domain associated with a small middle domain that contains the linker-peptide, followed by the receptor-binding domain. In contrast to prototypical single-chain AB toxins, AIP56 does not comprise a typical structurally complex translocation domain; instead, the elements involved in translocation are scattered across its domains. Thus, the catalytic domain contains a helical hairpin that serves as a molecular switch for triggering the conformational changes necessary for membrane insertion only with endosomal acidification, whereas the middle and receptor-binding domains are required for pore formation. Considering the increasing number of putative AIP56 homologous toxins in databases, the data presented here have implications far beyond the AIP56 intoxication process and the virulence ofPhdp.

Background Transcranial direct current stimulation (tDCS) provides a noninvasive polarity-specific constant current to treat epilepsy, through a mechanism possibly involving excitability modulation and neural oscillation. Objective To determine whether EEG oscillations underlie the interictal spike changes after tDCS in rats with chronic spontaneous seizures. Methods Rats with kainic acid-induced spontaneous seizures were subjected to cathodal tDCS or sham stimulation for 5 consecutive days. Video-EEG recordings were collected immediately pre- and post-stimulation and for the subsequent 2 weeks following stimulation. The acute pre-post stimulation and subacute follow-up changes of interictal spikes and EEG oscillations in tDCS-treated rats were compared with sham. Ictal EEG with seizure behaviors, hippocampal brain-derived neurotrophic factor (BDNF) protein expression, and mossy fiber sprouting were compared between tDCS and sham rats. Results Interictal spike counts were reduced immediately following tDCS with augmented delta and diminished beta and gamma oscillations compared with sham. Cathodal tDCS also enhanced delta oscillations in normal rats. However, increased numbers of interictal spikes with a decrease of delta and theta oscillations were observed in tDCS-treated rats compared with sham during the following 2 weeks after stimulation. Resuming tDCS suppressed the increase of interictal spike activity. In tDCS rats, hippocampal BDNF protein expression was decreased while mossy fiber sprouting did not change compared with sham. Conclusions The inverse relationship between the changes of delta oscillation and interictal spikes during tDCS on and off stimulation periods indicates that an enhanced endogenous delta oscillation underlies the tDCS inhibitory effect on epileptic excitability.

International audience; The Mekong River is one of the largest rivers in the world and hosts the second greatest fish diversity in the world after the Amazon. However, despite the importance of this diversity and its associated biomass for human food security and the economy, different anthropogenic pressures threaten the sustainability of the Mekong River and fish diversity, including the intense damming of the main river. Both the increase in salt-water penetration into the Mekong Delta and the disrupted connectivity of the river may have serious impacts on the numerous freshwater and migratory species. To evaluate the potential of an eDNA approach for monitoring fish diversity, water was sampled at 15 sites along the salinity gradient in the Mekong Delta and along 1500 km of the main stream, from Vietnam to Thailand and Laos. A total of 287 OTUs were recovered, of which 158 were identified to the species level using both reference sequences available in GenBank and references obtained locally. Agglomerative hierarchical clustering and PCA identified up to three main species assemblages in our samples. If the transition from brackish to freshwater conditions represents the main barrier between two of these assemblages, more surprisingly, the two other assemblages were observed in the freshwater Mekong, with a spatial disjunction that did not match any biogeographic ecoregion or the Khone falls, the latter thought to be an important fish dispersion barrier. Between 60% and 95% of the freshwater species were potamodromous. This pioneer eDNA study in the Mekong River at this geographical and ecological scale clearly confirmed the potential of this approach for ecological and diversity monitoring. It also demonstrated the need to rapidly build an exhaustive Mekong fish barcode library to enable more accurate species’ assignment. More eDNA surveys can now be expected to better describe the ecological niche of different species, which is crucial for any models aimed at predicting the impact of future damming of the Mekong.

There is a need for an automated morphometry algorithm to facilitate the otherwise labor-intensive task of the quantitative histological analysis of neural microscopic images. A benchmark morphometry algorithm is the convolutional neural network Axondeepseg (ADS), which yields a high segmentation accuracy for scanning and transmission electron microscopy images. Nevertheless, it shows decreased accuracy when applied to optical microscopy images, and it has been observed to yield sizable false positives when identifying small-sized neurons within the slides. In this study, ensemble learning is used to enhance the performance of ADS by combining it with the paired image-to-image translation algorithm PairedImageTranslation (PIT). Here, 120 optical microscopy images of peripheral nerves were used to train and test the ensemble learning model and the two base models individually for comparison. The results showed weighted pixel-wise accuracy for the ensemble model of 95.5%, whereas the ADS and PIT yielded accuracies of 93.4% and 90%, respectively. The automated measurements of the axon diameters and myelin thicknesses from the manually marked ground truth images were not statistically different (p = 0.05) from the measurements taken from the same images when segmented using the developed ensemble model, while they were different when they were measured from the segmented images by the two base models individually. The automated measurement of the G ratios indicated a higher similarity to the ground truth testing images for the ensemble model in comparison with the individual base models. The proposed model yielded automated segmentation of the nerve slides, which were sufficiently equivalent to the manual annotations and could be employed for axon diameters and myelin thickness measurements for fully automated histological analysis of the neural images.

Mon objet d’etude s’appuie sur le dispositif academique « Aller bien pour Mieux Apprendre » (ABMA). La demarche ABMA consiste a mettre en œuvre une dynamique au niveau de l’ensemble des etablissements scolaires, afin d’ameliorer le climat scolaire et favoriser la reussite de tous. Les differentes modalites d’accompagnement permettent aux etablissements de s’approprier la demarche et consistent a amener les etablissements scolaires a prendre en compte la sante et le bien-etre des eleves et de l’ensemble de la communaute educative. Cette recherche met en evidence le necessaire renouvellement des pratiques au sein des etablissements, afin de prendre en compte le « mieux-etre » et le « mieux-vivre ensemble » de l’ensemble de la communaute educative favorisant la reussite de tous. Cette recherche me permet d’affirmer l’importance d’un accompagnement effectue par des referents departementaux (professionnels de l’Education nationale) au sein des etablissements pour permettre la mise en œuvre de la demarche ABMA et pour initier et favoriser le changement. Il en resulte que les modalites d’accompagnement s’adaptent aux etablissements et sont en lien avec les competences propres developpees par les referents departementaux.

International audience; To enable chromosomal replication, DNA is unwound by the ATPase molecular motor replicative helicase. The bacterial helicase DnaB is a ring-shaped homo-hexamer whose conformational dynamics are being studied through its different 3D structural states adopted along its functional cycle. Our findings describe a new crystal structure for the apo-DnaB from Vibrio cholerae, forming a planar hexamer with pseudo-symmetry, constituted by a trimer of dimers in which the C-terminal domains delimit a triskelion-shaped hole. This hexamer is labile and inactive. We suggest that it represents an intermediate state allowing the formation of the active NTP-bound hexamer from dimers.

WOS:000552346200001; The almaco jack, Seriola rivoliana, is a circumtropical pelagic fish of importance both in commercial fisheries and in aquaculture. To understand levels of genetic diversity within and among populations in the wild, population genetic structure and the relative magnitude of migration were assessed using mtDNA sequence data and single nucleotide polymorphisms (SNPs) from individuals sampled from locations in the Pacific and Atlantic Oceans. A total of 25 variable sites of cytochrome c oxidase subunit 1 and 3678 neutral SNPs were recovered. Three genetic groups were identified, with both marker types distributed in different oceanic regions: Pacific-1 in central Pacific, Pacific-2 in eastern Pacific and Atlantic in western Atlantic. Nonetheless, the analysis of SNP identified a fourth population in the Pacific coast of Baja California Sur, Mexico (Pacific-3), whereas that of mtDNA did not. This mito-nuclear discordance is likely explained by a recently diverged Pacific-3 population. In addition, two mtDNA haplogroups were found within the western Atlantic, likely indicating that the species came into the Atlantic from the Indian Ocean with historical gene flow from the eastern Pacific. Relative gene flow among ocean basins was low with rm \textless 0.2, whereas in the eastern Pacific it was asymmetric and higher from south to north (rm \textgreater 0.79). The results reflect the importance of assessing genetic structure and gene flow of natural populations for the purposes of sustainable management.

Calmodulin (CaM) is a eukaryotic multifunctional, calcium-modulated protein that regulates the activity of numerous effector proteins involved in a variety of physiological processes. Calmidazolium (CDZ) is a potent small molecule antagonist of CaM and one the most widely used inhibitors of CaM in cell biology. Here, we report the structural characterization of CaM:CDZ complexes using combined SAXS, X-ray crystallography, HDX-MS and NMR approaches. Our results provide molecular insights into the CDZ-induced dynamics and structural changes of CaM leading to its inhibition. CDZ-binding induces an open-to-closed conformational change of CaM and results in a strong stabilization of its structural elements associated with a reduction of protein dynamics over a large time range. These CDZ-triggered CaM changes mimic those induced by CaM-binding peptides derived from protein targets, despite their distant chemical nature. CaM residues in close contact with CDZ and involved in the stabilization of the CaM:CDZ complex have been identified. These results open the way to rationally design new CaM-selective drugs.Figure and text for the Table of Contents (ToC)Calmidazolium is a potent and widely used inhibitor of calmodulin, a major mediator of calcium-signaling in eukaryotic cells. Structural characterization of calmidazolium-binding to calmodulin reveals that it triggers open-to-closed conformational changes similar to those induced by calmodulin-binding peptides derived from enzyme targets. These results open the way to rationally design new and more selective inhibitors of calmodulin.

The vagus nerve is the largest autonomic nerve and a major target of stimulation therapies for a wide variety of chronic diseases. However, chronic recording from the vagus nerve has been limited, leading to significant gaps in our understanding of vagus nerve function and therapeutic mechanisms. In this study, we use a carbon nanotube yarn (CNTY) biosensor to chronically record from the vagus nerves of freely moving rats for over 40 continuous hours. Vagal activity was analyzed using a variety of techniques, such as spike sorting, spike-firing rates, and interspike intervals. Many spike-cluster-firing rates were found to correlate with food intake, and the neural-firing rates were used to classify eating and other behaviors. To our knowledge, this is the first chronic recording and decoding of activity in the vagus nerve of freely moving animals enabled by the axon-like properties of the CNTY biosensor in both size and flexibility and provides an important step forward in our ability to understand spontaneous vagus-nerve function.

Indispensable dans l’exercice de notre metier, la formation professionnelle continue est definie comme un ensemble d’actions ayant pour objet de favoriser l’insertion professionnelle, le developpement des competences, l’acces aux niveaux de qualification professionnelle et la securisation des parcours professionnels. Inscrit dans la loi des 1966 (code du travail, sixieme partie), d’abord limite aux seuls salaries puis etendu ulterieurement aux travailleurs independants, le droit a la format...

Replicative helicases are essential proteins that unwind DNA in front of replication forks. Their loading depends on accessory proteins and in bacteria, DnaC and DnaI are well characterized loaders. However, most bacteria do not express either of these two proteins. Instead, they are proposed to rely on DciA, an ancestral protein unrelated to DnaC/I. While the DciA structure from Vibrio cholerae shares no homology with DnaC, it reveals similarities with DnaA and DnaX, two proteins involved during replication initiation. As other bacterial replicative helicases, VcDnaB adopts a toroid-shaped homo-hexameric structure, but with a slightly open dynamic conformation in the free state. We show that VcDnaB can load itself on DNA in vitro and that VcDciA stimulates this function, resulting in an increased DNA unwinding. VcDciA interacts with VcDnaB with a 3/6 stoichiometry and we show that a determinant residue, which discriminates DciA- and DnaC/I-helicases, is critical in vivo. Our work is the first step toward the understanding of the ancestral mode of loading of bacterial replicative helicases on DNA. It sheds light on the strategy employed by phage helicase loaders to hijack bacterial replicative helicases and may explain the recurrent domestication of dnaC/I through evolution in bacteria., Graphical Abstract Graphical AbstractDciA is the antecedent but unrelated to the helicase loaders DnaC/I. Structurally, the N-terminal domain of DciA is related to the NTD of DnaA, whereas its CTD is unfolded. DciA associates with DnaB according to a 3/6 stoichiometry which loads the helicase more efficiently on DNA. We identified in DnaB proteins a determinant residue that discriminates DciA- and DnaC/I-helicases and which happens to be critical in vivo. The determinant residue is located at the center of the DciA binding site.

It is well documented that synapses play a significant role in the transmission of information between neurons. However, in the absence of synaptic transmission, neural activity has been observed to continue to propagate. Previous studies have shown that propagation of epileptiform activity takes place in the absence of synaptic transmission and gap junctions and is outside the range of ionic diffusion and axonal conduction. Computer simulations indicate that electric field coupling could be responsible for the propagation of neural activity under pathological conditions such as epilepsy. Electric fields can modulate neuronal membrane voltage, but there is no experimental evidence suggesting that electric field coupling can mediate self-regenerating propagation of neural activity. Here we examine the role of electric field coupling by eliminating all forms of neural communications except electric field coupling with a cut through the neural tissue. We show that 4-AP induced activity generates an electric field capable of recruiting neurons on the distal side of the cut. Experiments also show that applied electric fields with amplitudes similar to endogenous values can induce propagating waves. Finally, we show that canceling the electrical field at a given point can block spontaneous propagation. The results from these in vitro electrophysiology experiments suggest that electric field coupling is a critical mechanism for non-synaptic neural propagation and therefore could contribute to the propagation of epileptic activity in the brain.

International audience; Phagocyte NADPH oxidase produces superoxide anions, a precursor of reactive oxygen species (ROS) critical for host responses to microbial infections. However, uncontrolled ROS production contributes to inflammation, making NADPH oxidase a major drug target. It consists of two membranous (Nox2 and p22phox) and three cytosolic subunits (p40, p47, and p67) that undergo structural changes during enzyme activation. Unraveling the interactions between these subunits and the resulting conformation of the complex could shed light on NADPH oxidase regulation and help identify inhibition sites. However, the structures and the interactions of flexible proteins comprising several well-structured domains connected by intrinsically disordered protein segments are difficult to investigate by conventional techniques such as X-ray crystallography, NMR, or cryo-EM. Here, we developed an analytical strategy based on FRET-fluorescence lifetime imaging (FLIM) and fluorescence cross-correlation spectroscopy (FCCS) to structurally and quantitatively characterize NADPH oxidase in live cells. We characterized the inter- and intramolecular interactions of its cytosolic subunits by elucidating their conformation, stoichiometry, interacting fraction, and affinities in live cells. Our results revealed that the three subunits have a 111 stoichiometry and that nearly 100% of them are present in complexes in living cells. Furthermore, combining FRET data with small-angle X-ray scattering (SAXS) models and published crystal structures of isolated domains and subunits, we built a 3D model of the entire cytosolic complex. The model disclosed an elongated complex containing a flexible hinge separating two domains ideally positioned at one end of the complex and critical for oxidase activation and interactions with membrane components.

The flathead grey mullet species (Mugil cephalus L.) Vietnam is an euryhaline fish whose distribution ranges from the North to the South. Currently, M. cephalus is facing the threat of overexploitation, and the number of individuals in most populations is declining fast. In order to conserve the species in Vietnam, it is necessary to evaluate its genetic diversity. Therefore, this study was carried out in Vietnam from August 2012 to June 2015. The results of our study provided the analysis for 12 locus microsatellites (SSR) from 70 mature individuals (standard length > 25 cm). The study also identified a total of 35 alleles for all loci studied, among which 10 loci were polymorphic. Average value of polymorphic information content (PIC) for each polymorphic marker was 0.2889 (0.0289 to 0.5918). Coefficient heterozygous gene Ho = 0.942; He = 0.517; Fst = 0.216; Fis = - 0.8211 (Fis < 0). The genetic relationship of 70 specimens or M. cephalus in Vietnam were divided in three main groups according to three specific geographic distributions in the country, the Gulf of Tonkin, the Central and the South. There were also mixed clusters observed among the three studied regions. Individuals in close geographical distance often clustered together and formed separate groups, in which the level of molecular changes were quite low, 29% among populations, and 71% among individuals within the same population.

The molecular mechanisms and forces involved in the translocation of bacterial toxins into host cells are still a matter of intense research. The adenylate cyclase (CyaA) toxin from Bordetella pertussis displays a unique intoxication pathway in which its catalytic domain is directly translocated across target cell membranes. The CyaA translocation region contains a segment, P454 (residues 454–484), which exhibits membrane‐active properties related to antimicrobial peptides. Herein, the results show that this peptide is able to translocate across membranes and to interact with calmodulin (CaM). Structural and biophysical analyses reveal the key residues of P454 involved in membrane destabilization and calmodulin binding. Mutational analysis demonstrates that these residues play a crucial role in CyaA translocation into target cells. In addition, calmidazolium, a calmodulin inhibitor, efficiently blocks CyaA internalization. It is proposed that after CyaA binding to target cells, the P454 segment destabilizes the plasma membrane, translocates across the lipid bilayer and binds calmodulin. Trapping of CyaA by the CaM:P454 interaction in the cytosol may assist the entry of the N‐terminal catalytic domain by converting the stochastic motion of the polypeptide chain through the membrane into an efficient vectorial chain translocation into host cells., The mechanism of cell invasion by bacterial toxins is still poorly understood. The adenylate cyclase (CyaA) toxin from Bordetella pertussis, the causative agent of whooping cough, directly translocates its catalytic domain across plasma membranes. The membrane‐permeabilizing segment (yellow) of CyaA translocates across the plasma membrane and binds calmodulin (red), which assists the entry of the catalytic domain (blue) into host cells while the hydrophobic and acylation domains (green) interact with the membrane and the C‐terminal Repeat‐in‐Toxin domain (grey) remains in the extra‐cellular milieu.

Objective One of the challenges in treating patients with drug-resistant epilepsy is that the mechanisms of seizures are unknown. Most current interventions are based on the assumption that epileptic activity recruits neurons and progresses by synaptic transmission. However, several experimental studies have shown that neural activity in rodent hippocampi can propagate independently of synaptic transmission. Recent studies suggest these waves are self-propagating by electric field (ephaptic) coupling. In this study, we tested the hypothesis that neural recruitment during seizures can occur by electric field coupling. Methods 4-Aminopyridine was used in both in vivo and in vitro preparation to trigger seizures or epileptiform activity. A transection was made in the in vivo hippocampus and in vitro hippocampal and cortical slices to study whether the induced seizure activity can recruit neurons across the gap. A computational model was built to test whether ephaptic coupling alone can account for neural recruitment across the transection. The model prediction was further validated by in vitro experiments. Results Experimental results show that electric fields generated by seizure-like activity in the hippocampus both in vitro and in vivo can recruit neurons locally and through a transection of the tissue. The computational model suggests that the neural recruitment across the transection is mediated by electric field coupling. With in vitro experiments, we show that a dielectric material can block the recruitment of epileptiform activity across a transection, and that the electric fields measured within the gap are similar to those predicted by model simulations. Furthermore, this nonsynaptic neural recruitment is also observed in cortical slices, suggesting that this effect is robust in brain tissue. Significance These results indicate that ephaptic coupling, a nonsynaptic mechanism, can underlie neural recruitment by a small electric field generated by seizure activity and could explain the low success rate of surgical transections in epilepsy patients.

Mgr Louis Petit, aujourd’hui bien oublie, a eu un destin hors du commun. Ne en Haute‑Savoie en 1868, deux ans seulement apres le rattachement a la France, dans un milieu pauvre, il entra dans la congregation des assomptionnistes apres avoir recu une formation assez superficielle, notamment en theologie. D’abord oriente vers un engagement de presse aupres de La Croix du Midi a Toulouse, il finit par devenir, grâce a une formation autodidacte etonnante, un veritable savant. Passionne par l’Orie...

ErbB2 (or HER2) is a receptor tyrosine kinase overexpressed in some breast cancers, associated with poor prognosis. Treatments targeting the receptor extracellular and kinase domains have greatly improved disease outcome in the last twenty years. In parallel, the structures of these domains have been described, enabling better mechanistic understanding of the receptor function and targeted inhibition. However, ErbB2 disordered C-terminal cytoplasmic tail (CtErbB2) remains very poorly characterized in terms of structure, dynamics and detailed functional mechanism. Yet, it is where signal transduction is triggered, via phosphorylation of tyrosine residues, and carried out, via interaction with adaptor proteins. Here we report the first description of ErbB2 disordered tail at atomic resolution, using NMR and SAXS. We show that although no part of CtErbB2 has any stable secondary or tertiary structure, it has around 20% propensity for a N-terminal helix that is suspected to interact with the kinase domain, and many PPII stretches distributed all along the sequence, forming potential SH3 and WW domains binding sites. Moreover, we identified a long-range transient contact involving CtErbB2 termini. These characteristics suggest new potential mechanisms of auto-regulation and protein-protein interaction.SIGNIFICANCEWe report here the first description of the receptor tyrosine kinase ErbB2 disordered tail (CtErbB2) at atomic resolution, using NMR and SAXS. We show that although CtErbB2 exhibits no stable structure, it does exhibit partial secondary and tertiary structures likely important for its function. These structural elements are consistent with an active role of the C-terminal tail in the regulation of the receptor’s activity, thanks to the presence of preformed structures for intramolecular interactions, as well as long-range contacts modulating accessibility of those sites and proline interaction sites distinct from the main tyrosine sites. Together, those results reinforce the view that disordered tails of receptors are more than random anchors for partners.

We present a model-independent method to reconstruct the impact parameter distributions of experimental data for intermediate energy heavy ion collisions, adapted from a recently proposed approach for ultra-relativistic heavy ion collisions. The method takes into account the fluctuations which are inherent to the relationship between any experimental observable and the impact parameter in this energy range. We apply the method to the very large dataset on heavy ion collisions in the energy range 20-100 MeV/nucleon obtained with the INDRA multidetector since 1993, for two observables which are the most commonly used for the estimation of impact parameters in this energy range. The mean impact parameters deduced with this new method for "central" collisions selected using typical observable cuts are shown to be significantly larger than those found when fluctuations are neglected, and as expected the difference increases as bombarding energy decreases. In addition, we will show that this new approach may provide previously inaccessible experimental constraints for transport models, such as an estimation of the extrapolated mean value of experimental observables for b = 0 collisions. The ability to give more realistic, model-independent, estimations of the impact parameters associated to different experimental datasets should improve the pertinence of comparisons with transport model calculations which are essential to better constrain the equation of state of nuclear matter., 14 pages, 7 figures. Revised version accepted for publication in Physical Review C

The SuperNEMO experiment will search for neutrinoless double-beta decay ($0\nu\beta\beta$), and study the Standard-Model double-beta decay process ($2\nu\beta\beta$). The SuperNEMO technology can measure the energy of each of the electrons produced in a double-beta ($\beta\beta$) decay, and can reconstruct the topology of their individual tracks. The study of the double-beta decay spectrum requires very accurate energy calibration to be carried out periodically. The SuperNEMO Demonstrator Module will be calibrated using 42 calibration sources, each consisting of a droplet of $^{207}$Bi within a frame assembly. The quality of these sources, which depends upon the entire $^{207}$Bi droplet being contained within the frame, is key for correctly calibrating SuperNEMO's energy response. In this paper, we present a novel method for precisely measuring the exact geometry of the deposition of $^{207}$Bi droplets within the frames, using Timepix pixel detectors. We studied 49 different sources and selected 42 high-quality sources with the most central source positioning., Comment: 16 pages, 12 figures, submitted to JINST, response to reviewer comments