Five Years Monitoring the Emergence of Unregulated Toxins in Shellfish in France (EMERGTOX 2018-2022).

Shellfish accumulate microalgal toxins, which can make them unsafe for human consumption. In France, in accordance with EU regulations, three groups of marine toxins are currently under official monitoring: lipophilic toxins, saxitoxins, and domoic acid. Other unregulated toxin groups are also present in European shellfish, including emerging lipophilic and hydrophilic marine toxins (e.g., pinnatoxins, brevetoxins) and the neurotoxin ß-N-methylamino-L-alanine (BMAA). To acquire data on emerging toxins in France, the monitoring program EMERGTOX was set up along the French coasts in 2018. Three new broad-spectrum LC-MS/MS methods were developed to quantify regulated and unregulated lipophilic and hydrophilic toxins and the BMAA group in shellfish (bivalve mollusks and gastropods). A single-laboratory validation of each of these methods was performed. Additionally, these specific, reliable, and sensitive operating procedures allowed the detection of groups of EU unregulated toxins in shellfish samples from French coasts: spirolides (SPX-13-DesMeC, SPX-DesMeD), pinnatoxins (PnTX-G, PnTX-A), gymnodimines (GYM-A), brevetoxins (BTX-2, BTX-3), microcystins (dmMC-RR, MC-RR), anatoxin, cylindrospermopsin and BMAA/DAB. Here, we present essentially the results of the unregulated toxins obtained from the French EMERGTOX monitoring plan during the past five years (2018-2022). Based on our findings, we outline future needs for monitoring to protect consumers from emerging unregulated toxins.

First Evidence of the Presence of Anatoxin-A in Sea Figs Associated with Human Food Poisonings in France.

From January 2011 to March 2018, 26 patients aged from 20 to 80 years old reported being sick in France after eating sea figs of the genus Microcosmus. The patients had symptoms evoking a cerebellar syndrome: blurred or double vision, ataxia and dizziness, asthenia, headache, muscle cramps, paresthesia and digestive disorders (nausea, vomiting and diarrhea). Three of the 18 food poisoning events recorded by the Poison Control Center in Marseille and involving four patients were further investigated as the meal leftovers were collected and analyzed. A previous study ruled out the presence of the regulated lipophilic marine toxins after high-resolution mass spectrometry, but further analyses were required to look for hydrophilic cyanotoxins. The sea fig leftovers from food poisoning case Numbers 1 (January 2011), 6 (December 2012) and 17 (March 2018) of this published case series were analyzed by hydrophilic interaction liquid chromatography coupled to low- and high-resolution mass spectrometry to investigate the presence of hydrophilic cyanotoxins. The sea fig samples showed anatoxin-a (ATX-a) concentrations ranging from 193.7 to 1240.2 µg/kg. The sea fig control sample analyzed was also contaminated with ATX-a but in a much smaller concentration (22.5 µg/kg). To the best of our knowledge, this is the first report of human food poisoning involving ATX-a as the possible causative toxin where the cyanotoxin could be unequivocally identified.

Synchrotron imaging of dentition provides insights into the biology of Hesperornis and Ichthyornis, the "last" toothed birds.

BACKGROUND: The dentitions of extinct organisms can provide pivotal information regarding their phylogenetic position, as well as paleobiology, diet, development, and growth. Extant birds are edentulous (toothless), but their closest relatives among stem birds, the Cretaceous Hesperornithiformes and Ichthyornithiformes, retained teeth. Despite their significant phylogenetic position immediately outside the avian crown group, the dentitions of these taxa have never been studied in detail. To obtain new insight into the biology of these 'last' toothed birds, we use cutting-edge visualisation techniques to describe their dentitions at unprecedented levels of detail, in particular propagation phase contrast x-ray synchrotron microtomography at high-resolution. RESULTS: Among other characteristics of tooth shape, growth, attachment, implantation, replacement, and dental tissue microstructures, revealed by these analyses, we find that tooth morphology and ornamentation differ greatly between the Hesperornithiformes and Ichthyornithiformes. We also highlight the first Old World, and youngest record of the major Mesozoic clade Ichthyornithiformes. Both taxa exhibit extremely thin and simple enamel. The extension rate of Hesperornis tooth dentine appears relatively high compared to non-avian dinosaurs. Root attachment is found for the first time to be fully thecodont via gomphosis in both taxa, but in Hesperornis secondary evolution led to teeth implantation in a groove, at least locally without a periodontal ligament. Dental replacement is shown to be lingual via a resorption pit in the root, in both taxa. CONCLUSIONS: Our results allow comparison with other archosaurs and also mammals, with implications regarding dental character evolution across amniotes. Some dental features of the 'last' toothed birds can be interpreted as functional adaptations related to diet and mode of predation, while others appear to be products of their peculiar phylogenetic heritage. The autapomorphic Hesperornis groove might have favoured firmer root attachment. These observations highlight complexity in the evolutionary history of tooth reduction in the avian lineage and also clarify alleged avian dental characteristics in the frame of a long-standing debate on bird origins. Finally, new hypotheses emerge that will possibly be tested by further analyses of avian teeth, for instance regarding dental replacement rates, or simplification and thinning of enamel throughout the course of early avian evolution.

Three-Dimensional Printed Knee Implants: Insights into Surgeons' Points of View.

Three-dimensional printing is a technology that has been developed and applied in several medical specialties, especially orthopedic surgery. Knee arthroplasty is the most commonly performed procedure. To fit the morphology of each knee, surgeons can choose between different standardized off-the-shelf implant sizes or opt for customized 3D-printed implants. However, routine adoption of the latter has been slow and faces several barriers. Existing studies focus on technical improvements or case studies and do not directly address the surgeon's perspective. Our study invited surgeons to express themselves freely and answer the question "What do you think about the manufacture of a prosthesis by 3D printing?". The questionnaire was completed by 90 surgeons. On average, they had more than 10 years of experience (52, 57.8% ± 10.2%), worked in public hospitals (54, 60% ± 10.1%), and performed between 0 and 100 prostheses per year (60, 66.7% ± 9.7%). They also reported not using planning software (47, 52.2% ± 9.7%), navigation systems, or robots (62, 68.9% ± 9.6%). Regarding the use of technological innovation, they agreed on the extra surgical time needed (67, 74.4% ± 9.0%). The answers obtained were classified according to two criteria: (i) opinions, and (ii) motivations. Among the respondents, 51 (70% ± 9.5%) had positive and 22 (30% ± 9.5%) had negative opinions about 3D printing. The motivations were distributed among seven categories (surgery, materials, costs, logistics, time, customization, and regulatory) and mainly related to "pre-surgery" and "post-surgery" concerns. Finally, the results showed that the use of navigation systems or robots may be associated with a more positive view of 3DP. The purpose of our study was to examine knee surgeons' perceptions of 3DP at a time of significant expansion of this technology. Our study showed that there was no opposition to its implementation, although some surgeons indicated that they were waiting for validated results. They also questioned the entire supply chain, including hospitals, insurance companies, and manufacturers. Although there was no opposition to its implementation, 3D printing is at a crucial point in its development and its full adoption will require advances in all areas of joint replacement.

Current Perspectives on Tooth Implantation, Attachment, and Replacement in Amniota.

Teeth and dentitions contain many morphological characters which give them a particularly important weight in comparative anatomy, systematics, physiology and ecology. As teeth are organs that contain the hardest mineralized tissues vertebrates can produce, their fossil remains are abundant and the study of their anatomy in fossil specimens is of major importance in evolutionary biology. Comparative anatomy has long favored studies of dental characters rather than features associated with tooth attachment and implantation. Here we review a large part of the historical and modern work on the attachment, implantation and replacement of teeth in Amniota. We propose synthetic definitions or redefinitions of most commonly used terms, some of which have led to confusion and conflation of terminology. In particular, there has long been much conflation between dental implantation that strictly concerns the geometrical aspects of the tooth-bone interface, and the nature of the dental attachment, which mostly concerns the histological features occurring at this interface. A second aim of this work was to evaluate the diversity of tooth attachment, implantation and replacement in extant and extinct amniotes in order to derive hypothetical evolutionary trends in these different dental traits over time. Continuous dental replacement prevails within amniotes, replacement being drastically modified only in Mammalia and when dental implantation is acrodont. By comparison, dental implantation frequently and rapidly changes at various taxonomic scales and is often homoplastic. This contrasts with the conservatism in the identity of the tooth attachment tissues (cementum, periodontal ligament, and alveolar bone), which were already present in the earliest known amniotes. Because the study of dental attachment requires invasive histological investigations, this trait is least documented and therefore its evolutionary history is currently poorly understood. Finally, it is essential to go on collecting data from all groups of amniotes in order to better understand and consequently better define dental characters.

Development and validation of an HPLC-MS/MS method with QuEChERS extraction using isotopic dilution to simultaneously analyze chlordecone and chlordecol in animal livers.

A QuEChERS extraction method followed by HPLC-MS/MS analysis was developed to simultaneously analyze chlordecone and its metabolite chlordecol in animal livers. The overall method was validated with accuracy profiles according to the French Standard NF V03-110 and European Union guidelines. The validation was performed on bovine, ovine and porcine liver samples. Linearity, matrix effect, accuracy, within-laboratory repeatability, specificity, LOQ, Q/q relative ion intensities, and uncertainty were reported. Recoveries were between 70% and 120%. LOQs of 1.36 µg chlordecone kg-1 and 2.50 µg chlordecol kg-1 of fresh liver were found. Twelve contaminated livers of bovine, ovine and porcine origin from the French West Indies or samples from in vivo studies were analyzed. In these liver samples from contaminated animals, chlordecone was quantified at concentrations higher than the maximum residue limit and chlordecol in very low amounts in all the samples. In addition, these results confirm that chlordecone can be metabolized in ruminant species.

Validation of analytical methods for chlordecone and its metabolites in the urine and feces of ewes.

Chlordecone (CLD) is a Persistent Organic Pollutant used between 1972 until 1993 in the French West Indies (FWI). Due to its persistence and extensive application, a quarter of the total local agricultural acreage is still moderate to heavily polluted. In consequence, livestock may be contaminated at various levels. This is a major public health concern, particularly for local consumers. In order to better understand the fate of CLD in livestock organisms, in vivo studies are required. There is no information available about its metabolism and elimination in ruminants, common livestock in the FWI. To be able to monitor the fate of chlordecone and its metabolites in livestock and to assess if the compounds could be released in the environment, urinary and fecal samples were logically targeted. In order to reach this goal, robust and validated analytical methods are required. For this purpose, Quick, Easy, Cheap, Effective, Rugged, and Safe (QuEChERS) extraction methods were validated to analyze CLD and its metabolites in the urine and feces. The analysis was carried using liquid phase chromatography with tandem mass spectrometry and validated according to French standard NF V03-110 and SANTE guidelines. Matrix effect, Accuracy, within-laboratory repeatability, specificity, Q/q relative ion intensities and uncertainty were reported. Recoveries between 70% and 120% were obtained from urine and feces. The limits of quantification (LOQ) in urine samples were 0.1 µg CLD L-1, 0.1 µg total CLD (CLD and its conjugates)·L-1, 1.3 µg CLDOH L-1 and 2.4 µg total CLD (chlordecol and its conjugates) L-1 of urine. LOQ in fresh feces were 3.2 µg CLD kg-1 and 5.8 µg CLDOH kg-1. Contaminated urinary and fecal samples from ewes were analyzed to confirm the relevance of the methods. In urine, CLD and conjugated CLDOH could be quantified whereas only free CLD and free CLDOH were found in feces. These methods are essential for future toxicokinetic studies and also to estimate the environmental contamination.

Right hemisphere dominance for auditory attention and its modulation by eye position: an event related fMRI study.

PURPOSE: Previous neuroimaging studies of oddball tasks and other paradigms measuring attention processes support right hemisphere dominance for attentional processes. Using an auditory selective attention task, we studied the functional asymmetry of the human brain in response to attended or unattended deviant tones. Secondly, we examined whether a congruency or a discrepancy between audio-spatial and visuo-spatial cued attentional resources may influence the activity elicited by an auditory selective attention task. METHODS: We used event-related functional magnetic resonance imaging (fMRI) to study healthy adults as they performed an auditory oddball task in which a spatial-cued instruction indicated the ear to attend a monaural deviant tone. We addressed the question of congruency/discrepancy between attentional resources by using three different eye positions during the performance of the auditory oddball task. RESULTS: Relative to standard tones, both attended and unattended deviant tones (DTs) presented to either ear elicit the activation of a widespread bilaterally distributed cortical and subcortical network. A subset of this network, essentially frontal and temporal areas, showed not only greater right than left activity but an enhancement of this rightward asymmetry in response to attended DTs. The only cortical region that showed a leftward asymmetry in response to attended DTs overlapped Heschl gyrus and planum temporale, unmasking a left hemisphere preference of both primary and secondary auditory cortex for processing simple attended monaural stimuli. Questioning the impact of eye position during auditory oddball task, we observed a lesser activity in right integrative crossmodal areas (superior temporal sulcus, opercular part of the inferior frontal gyrus, pre-SMA) when the eye positions were contralateral to detected DTs. These regions may be tuned to best respond when both visuo-spatial and audio-spatial attentional resources work together. CONCLUSION: These results support the assumption that the right hemisphere is preferentially engaged in processing audio-spatial attentional resources and underline the interest to study the crossmodal integration of attentional resources by the mean of the detection of DTs in different eye positions.