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Hybridization opens a unique window for observing speciation mechanisms and is a potential engine of speciation. One controversially discussed outcome of hybridization is homoploid hybrid speciation by reciprocal sorting, where a hybrid population maintains a mixed combination of the parental genetic incompatibilities, preventing further gene exchange between the newly formed population and the two parental sources. Previous work showed that, for specific linkage architectures (i.e., the genomic location and order of hybrid incompatibilities), reciprocal sorting could reliably result in hybrid speciation. Yet, the sorting of incompatibilities creates a risk of population extinction. To understand how demographic consequences of the purging of incompatibilities interact with the formation of a hybrid species, we model an isolated hybrid population resulting from a single admixture event. We study how population size, linkage architecture and the strength of the incompatibility affect survival of the hybrid population, resolution/purging of the genetic incompatibilities and the probability of observing hybrid speciation. We demonstrate that the extinction risk is highest for intermediately strong hybrid incompatibilities. In addition, the linkage architecture displaying the highest hybrid speciation probabilities changes drastically with population size. Overall, this indicates that population dynamics can strongly affect the outcome of hybridization and the hybrid speciation probability.

Natural hybrid zones offer a powerful framework for understanding the genetic basis of speciation in progress because ongoing hybridization continually creates unfavorable gene combinations. Evidence indicates that postzygotic reproductive isolation is often caused by epistatic interactions between mutations in different genes that evolved independently of one another (hybrid incompatibilities). We examined the potential to detect epistatic selection against incompatibilities from genome sequence data using the site frequency spectrum (SFS) of polymorphisms by conducting individual-based simulations in SLiM. We found that the genome-wide SFS in hybrid populations assumes a diagnostic shape, with the continual input of fixed differences between source populations via migration inducing a mass at intermediate allele frequency. Epistatic selection locally distorts the SFS as non-incompatibility alleles rise in frequency in a manner analogous to a selective sweep. Building on these results, we present a statistical method to identify genomic regions containing incompatibility loci that locates departures in the local SFS compared with the genome-wide SFS. Cross-validation studies demonstrate that our method detects recessive and codominant incompatibilities across a range of scenarios varying in the strength of epistatic selection, migration rate, and hybrid zone age. Our approach takes advantage of whole genome sequence data, does not require knowledge of demographic history, and can be applied to any pair of nascent species that forms a hybrid zone.

Organismal elemental stoichiometry is important at all scales of ecological interactions, particularly in symbiosis. Symbiotic partnerships are found extensively in corals, where coral hosts and their photosynthetic dinoflagellate partners trade essential nutrients. Using an ecological stoichiometry framework, we assessed variations in carbon (C), nitrogen (N), and phosphorus (P) concentrations in coral hosts and symbiotic zooxanthellae. Our aim was to assess whether corals and zooxanthellae differ in the stoichiometry of C, N and P, which may indicate that corals regulate their symbionts by restricting their access to nutrients. We also investigated the influence of biological and environmental factors on the elemental stoichiometry of coral–zooxanthellae symbiosis. Analysing field data from four locations on the Great Barrier Reef using boosted regression tree models, we observed that C:P and N:P ratios are significantly higher in zooxanthellae than in their coral hosts, suggesting P limitation for zooxanthellae, although the degree of P or N limitation in zooxanthellae varied among coral taxa. Coral taxonomy was also a significant driver of coral–zooxanthellae stoichiometry variations at the species level. Geographical location had a small influence on the variations in stoichiometry. Our analyses suggest that coral species may differentially control the access of zooxanthellae to nutrients which may lead to variation in reef coral responses to nutrient enrichment.

Hybridization has broad evolutionary consequences, from fueling or counteracting speciation to facilitating adaptation to novel environments. Hybridization and subsequent introgression appear widespread along the tree of life. However, our understanding of how distinct evolutionary forces shape admixed genomes and the fate of introgressed genetic variants remains scarce. Most admixture research in animals has focused on diploid organisms. We propose that haplodiploid organisms can help resolve open questions about the genomic consequences of hybridization in natural populations. The ploidy difference between haploid males and diploid females, the availability of genome-wide male haplotypes, and ongoing cases of admixture make haplodiploid organisms promising models to improve our knowledge with regards to the evolution of hybrid genomes.

Water transfer projects are effective measures to solve the uneven distribution of water resources in China. However, these projects create highways for Golden Mussels (Limnoperna fortunei), which are aquatic invasive species, to invade new habitats. This has caused a series of problems such as structure corrosion, enhanced flow resistance, pipe blocking, and water pollution. In order to design remediation strategies, it is important to understand the factors influencing the Golden Mussel invasion. This paper investigates the invasion of Golden Mussels in the East River Water Source Project and the Northern Inter-basin Water Transfer project in China. The maximum densities of Golden Mussel attachment in these projects are about 10 000 ind. m−2. Flow velocity was found to be a significant influencing factor: the optimal flow velocity for attachment is in the range of 0.3 to 0.9 m s−1. Water temperature was also an important factor: the water temperature in the cold season influences the density of Golden Mussels and additionally, their growth and shell size are related to food competition between different individuals and water temperature. There was no obvious relationship between the pressure and the attachment density of Golden Mussels. Because all data was obtained during an infrequent pause in the water transfer projects, the data of this study is limited and the conclusions are preliminary descriptive observations. For future management decisions, more rigorous studies should be conducted to develop quantitative models aimed at providing a confident basis for controlling Golden Mussel invasion in water transfer projects.

Medical problems related to illicit drug use are frequently encountered at electronic dance music (EDM) events. In this prospective study, the medical problems and toxicological analyses on intoxicated persons and seized materials are described jointly. The aim of this study is to find out to what extent these efforts may assist in developing prevention strategies and organising on-site care at EDM events. The most frequently encountered clinical presentation in the 121 included patients was: agitation/ aggression (26%), drunkenness (25%), depressed level of consciousness (24%) and hallucinations (9%). Only five patients were transported by ambulance to a hospital. In 100 of the 121 included patients (83%) an ethanolemia of at least 0.50 g/L was measured (with ethanol as the only drug found in 47 cases). 3,4-methylenedioxymethamphetamine (MDMA) was detected in 54% of the blood samples, cocaine in 11%, gamma-hydroxybutyric acid (GHB) in 11%, amphetamine in 7%, ketamine in 6% and a new psychoactive substance (NPS) in 4%. Except for 8 MDMA-users poly drug use was found in all these cases. The 178 seized samples most frequently contained MDMA (31%), cannabis (28%) or no active substance (15%). In 11 samples (6%) an NPS was detected. Of particular interest was a tablet containing 4-chloromethamphetamine (a previously unknown neurotoxic NPS), 4-chloroamphetamine, para-methoxyamphetamine, para-methoxymethamphetamine and ethylone. Our data show that at EDM events ethanol and MDMA are still the party drugs causing most health hazards and that NPS only play a minor role. Regarding the toxicological efforts, we recommend to analyse all seized materials from an EDM event, but only blood samples from the most severely intoxicated patients. (C) 2019 Elsevier B.V. All rights reserved.

International audience; Reef ecosystems are under increasing pressure from global and local stressors. Rising seawater temperature and high ultraviolet radiation (UVR) levels are the main drivers of the disruption of the coral-dinoflagellate symbiosis (bleaching). Bleaching can also be exacerbated by nitrate contamination in coastal reefs. However, the underlying physiological mechanisms are still poorly understood. Here, we assessed the physiological and oxidative state of the scleractinian coral Pocillopora damicornis, maintained eight weeks in a crossed-factorial design including two temperatures (26 °C or 30 °C), and two nitrate (0.5 and 3 μM-enriched), and UVR (no UVR and 25/1.5 Wm−2 UVA/B) levels. Nitrate enrichment, and high temperature, significantly impaired coral photosynthesis. However, UVR alleviated the nitrate and temperature-induced decrease in photosynthesis, by increasing the coral's antioxidant capacity. The present study contributes to our understanding of the combined effects of abiotic stressors on coral bleaching susceptibility. Such information is urgently needed to refine reef management strategies.

A key step in understanding the genetic basis of different evolutionary outcomes (e.g., adaptation) is to determine the roles played by different mutation types (e.g., SNPs, translocations and inversions). To do this we must simultaneously consider different mutation types in an evolutionary framework. Here, we propose a research framework that directly utilizes the most important characteristics of mutations, their population genetic effects, to determine their relative evolutionary significance in a given scenario. We review known population genetic effects of different mutation types and show how these may be connected to different evolutionary outcomes. We provide examples of how to implement this framework and pinpoint areas where more data, theory and synthesis are needed. Linking experimental and theoretical approaches to examine different mutation types simultaneously is a critical step towards understanding their evolutionary significance.

Non-alcoholic fatty liver disease represents the most common liver disease and is characterized by an excess of lipid accumulation in hepatocytes, mainly stored as triglycerides. Phaeodactylum tricornutum is a marine microalga, which is rich in bioactive molecules known to be hepatoprotective, such as n-3 long-chain polyunsaturated fatty acids and fucoxanthin. The aim of this study was to investigate the effects of a carotenoid extract from P. tricornutum in a cellular model of non-alcoholic fatty liver disease induced by palmitate treatment. The combined effects of carotenoids and lipids, especially n-3 long-chain polyunsaturated fatty acids, were also investigated by using a total lipophilic extract. HepG2 cells were exposed for 24 h to 250 &micro, M palmitate with or without the addition of carotenoid extract (6 &mu, g/mL) or total lipophilic extract (100 &mu, g/mL). The addition of carotenoid extract or total lipophilic extract prevented the accumulation of triglycerides, total cholesterol and cholesterol esters. The carotenoid extract and total lipophilic extract also decreased the mRNA expression levels of genes involved in lipogenesis (ACACA, FASN, SCD and DGAT1) and cholesterol esterification (ACAT1/SOAT1). In addition, the total lipophilic extract also downregulated the LXR/NR1H3 and SREBF1 genes, which are involved in lipogenesis regulation. By contrast, the carotenoid extract increased the mRNA level of CPT1A, a &beta, oxidation related gene, and reduced the lipid droplet accumulation. In conclusion, this study highlights the preventive effects against non-alcoholic fatty liver disease of the two microalga extracts.

Despite considerable efforts in prevention and therapy, breast cancer remains a major public health concern worldwide. Numerous studies using breast cancer cell lines have shown the antiproliferative and pro-apoptotic effects of docosahexaenoic acid (DHA). Some studies have also demonstrated the inhibitory effect of DHA on the migration and invasion of breast cancer cells, making DHA a potential anti-metastatic agent. Thus, DHA has shown its potential as a chemotherapeutic adjuvant. However, the molecular mechanisms triggering DHA effects remain unclear, and the aim of this study was to provide a transcriptomic basis for further cellular and molecular investigations. Therefore, MDA-MB-231 cells were treated with 100 &micro, M DHA for 1`2 h or 24 h before RNA-seq analysis. The results show the great impact of DHA-treatment on the transcriptome, especially after 24 h of treatment. The impact of DHA is particularly visible in genes involved in the cholesterol biosynthesis pathway that is strongly downregulated, and the endoplasmic reticulum (ER)-stress response that is, conversely, upregulated. This ER-stress and unfolded protein response could explain the pro-apoptotic effect of DHA. The expression of genes related to migration and invasion (especially SERPINE1, PLAT, and MMP11) is also impacted by DHA. In conclusion, this transcriptomic analysis supports the antiproliferative, pro-apoptotic and anti-invasive effects of DHA, and provides new avenues for understanding its molecular mechanisms.

« Une lente et régulière évolution » Sans doute s’étonnera-t-on de voir la linguistique des années 1860-1870 saisie par le darwinisme. N’ayons garde toutefois d’en blâmer l’incongruité. Le parallèle des langues et des espèces, jugé à l’époque « remarquable » et réel, souligne surtout la conversion du siècle au paradigme historique. Le Développement, affirmait Robert Chambers, est la « seule loi » du monde organique et toutes les manifestations de l’esprit s’y soumettent comme elles l’exprimen...

La théorie de Darwin aussi bien que sa réception scientifique, littéraire et politique en France continuent d’être matière à débat. Les controverses qu’elle a suscitées à partir de la première traduction française de L’Origine des espèces en 1862 n’ont pas été le seul fait des naturalistes, des géologues ou des paléontologues. De nombreux autres savants se sont préoccupés de l’impact du darwinisme sur leur discipline, notamment en philologie, dans les mathématiques, la linguistique, la psychologie comparée, l’histoire ou la philosophie. Les auteurs des textes réunis dans Darwin au Collège de France montrent que le Collège de France constitue un point de référence idéal pour étudier les idées de Darwin et leur réception ainsi que la manière dont elles ont pu jouer sur la formation des disciplines du milieu du xixe siècle jusqu’à aujourd’hui. Parmi les défenseurs ou les adversaires français du darwinisme, on compte en effet plusieurs de ses professeurs, à commencer par Pierre Flourens, dont l’Examen du livre de M. Darwin (1864) a été l’un des premiers comptes rendus hostiles de L’Origine des espèces. D’autres figurent encore dans la liste : Ernest Renan, Edgar Quinet, Théodule Ribot, Jean-Charles Lévêque, Étienne-Jules Marey, Jean-François Nourrisson, Michel Bréal, Gabriel Tarde, Henri Bergson, Étienne Gilson. À cette liste s’ajoutent les professeurs de biologie, de médecine, de paléontologie, de neurosciences, ou d’économie politique. Le colloque, dont les contributions sur quelques-uns des savants mentionnés ci-dessus sont réunies dans le deuxième volume de la collection « Passage des disciplines », a été organisé par Alain Prochiantz et Antoine Compagnon dans le cadre du projet « Passage des disciplines : Histoire globale du Collège de France, xixe siècle - xxe siècle », qui s’intéresse à l’évolution des matières enseignées aussi bien que celles qui ont n’y ont pas été admises et qui forment un « Collège virtuel », depuis la fin du xviiie siècle jusqu’aux années 1960. Il est dirigé par Antoine Compagnon, avec la collaboration de Céline Surprenant et le soutien financier de PSL (2016-2019), et de la Fondation Hugot. The controversies that Darwin’s theories sparked off from the first French translation of The Origins of Species in 1862 onwards were not the doing of naturalists, geologists or paleontologists alone. Many other scientists and scholars were likewise concerned about the impact of Darwinism on their fields of knowledge. The authors of the collection of texts in Darwin au Collège de France show that the Collège de France is an ideal point of reference for studying Darwin’s ideas and their reception, particularly the way in which they impacted on the organization of scientific disciplines from the middle of the 19th century onwards. Many professors of the Collège de France were among the advocates or the adversaries of Darwinism, starting with Pierre Flourens, whose Examen du livre de M. Darwin (1864) was one of the first negative reviews of The Origins of Species in France. The list includes Ernest Renan, Edgar Quinet, Théodule Ribot, Jean-Charles Lévêque, Étienne-Jules Marey, Jean-François Nourrisson, Michel Bréal, Gabriel Tarde, Henri Bergson, and Étienne Gilson, in addition to professors of biology, medicine, neurosciences, or political economy. Volume 2 of the “Passage des disciplines” series includes texts presented at a symposium organized by Alain Prochiantz and Antoine Compagnon as part of “Passage des disciplines: Global history of the Collège de France, 19th-20th century”. The series examines the genesis of scientific and literary fields of knowledge, their mapping and their evolution during the 19th and 20th century, on both national international levels, from the standpoint afforded by the history of the renewal of academic Chairs at the Collège de France. This research programme is headed by Antoine Compagnon, in collaboration with Céline Surprenant, and receives the financial support of PSL (2016-2019) and the Fondation Hugot.

Parapatric speciation has recently received a lot of attention. By encompassing the whole continuum between allopatric and sympatric scenarios, it includes many potential scenarios for the evolution of new species. Building upon previous work, we investigate how a genetic barrier to gene flow, that relies on a single postzygotic genetic incompatibility, may further evolve. We consider a continent island model with three loci involved in pairwise Dobzhansky-Muller incompatibilities (DMIs). Using a deterministic and analytic approach, we derive the conditions for invasion of a new mutation and its consequences on an already existing genetic barrier to gene flow. We focus on quantifying the impact of the epistasis generated by the new mutation on the genetic barrier. We show that the accumulation of genetic incompatibilities in the presence of gene flow is a complex process, where new mutations can either strengthen or destroy a preexisting barrier. In particular, preexisting polymorphism and incompatibilities do not always facilitate the growth of the genetic barrier by accumulation of further barrier genes. Migration may disrupt the snowball effect (the accelerating rate of DMI accumulation in allopatry) because incompatibilities are directly tested by selection. Our results also show an ambiguous role of gene flow, which can either impede or facilitate the strengthening of the genetic barrier. Overall, our results illustrate how the inclusion of gene flow renders the building of a genetic barrier difficult to analyze.

Nitrogen (N) and phosphorus (P) limitations induce triacylglycerol (TAG) accumulation and membrane lipid remodelling in the marine diatom Phaeodactylum tricornutum. However, a clear understanding of the metabolic reorientation is still lacking. Carbon partitioning is of great interest because this microalga produces various highly valuable molecules such as lipids and polyunsaturated fatty acids. This study compared growth, photosynthetic activity, biochemical and transcriptional responses of P. tricornutum throughout batch culture under N or P limitation. The integrated results show that the photosynthetic intensity was greatly reduced under N or P limitation. Under N limitation, the degradation and re-use of cellular N-containing compounds contributed to TAG accumulation, whilst P limitation favoured TAG accumulation due to the efficiency of carbon fixation, without massive degradation of essential compounds at cellular level. There was no difference in the partitioning of carbon to neutral lipids between N and P limitation. Substitution of phospholipids with betaine lipids appeared to be a P-specific acclimation strategy in P. tricornutum, which was largely regulated at the gene expression level. Betaine lipid synthesis was induced by P limitation. The lipid remodelling began once the medium became deficient in P. While the phospholipid biosynthesis pathway was not completely inhibited, a shift of lipid classes occurred immediately after their synthesis via phospholipid-recycling mechanisms.

This study aims to employ a pulsed electric field (PEF) treatment for the biocompatible (non-destructive) extraction of proteins from living cells of the green microalga Haematococcus pluvialis. Using a field strength of 1 kV cm-1, we achieved the extraction of 10.2 µg protein per mL of culture , which corresponded to 46% of the total amount of proteins that could be extracted by complete destructive extraction (i.e. the grinding of biomass with glass beads). We found that the extraction yield was not improved by stronger field strengths and was not dependent on the pulse frequency. A biocompatibility index (BI) was defined as the relative abundance of cells that remained alive after the PEF treatment. This index relied on measurements of several physiological parameters after a PEF treatment. This way found that at 1 kV cm-1 that cultures recovered after 72 h. Therefore, these PEF conditions constituted a good compromise between protein extraction efficiency and culture survival. To characterize the PEF treatment further at a molecular level, mass spectrometry-based proteomics analyses of PEF-prepared extracts was used. This led to the identification of 52 electro-extracted proteins. Of these, only 16 proteins were identified when proteins were extracted with PEF at 0.5 cm-1. They belong to core metabolism, stress response and cell movement. Unassigned proteins were also extracted. Their physiological implications and possible utilization in food as alimentary complements are discussed.

Hybridization has recently gained considerable interest both as a unique window for observing speciation mechanisms and as a potential engine of speciation. The latter remains a controversial topic. It was recently hypothesized that the reciprocal sorting of genetic incompatibilities from parental species could result in hybrid speciation, when the hybrid population maintains a mixed combination of the parental incompatibilities that prevents further gene exchange with both parental populations. However, the specifics of the purging/sorting process of multiple incompatibilities have not been examined theoretically.We here investigate the allele-frequency dynamics of an isolated hybrid population that results from a single hybridization event. Using models of 2 or 4 loci, we investigate the fate of one or two genetic incompatibilities of the Dobzhansky-Muller type (DMIs). We study how various parameters affect both the sorting/purging of the DMIs and the probability of observing hybrid speciation by reciprocal sorting. We find that the probability of hybrid speciation is strongly dependent on the linkage architecture (i.e. the order and recombination distance between loci along chromosomes), the population size of the hybrid population, and the initial relative contribution of the parental populations to the hybrid population. We identify a Goldilocks zone for specific linkage architectures and intermediate recombination rates, in which hybrid speciation becomes highly probable. Whereas an equal contribution of parental populations to the hybrid populations maximizes the hybrid speciation probability in the Goldilocks zone, other linkage architectures yield unintuitive asymmetric maxima. We provide an explanation for this pattern, and discuss our results both with respect to the best conditions for observing hybrid speciation in nature and their implications regarding patterns of introgression in hybrid zones.SummaryHybridization is observed ubiquitously in nature. Its outcome can range from extinction to the creation of new species. With respect to the latter, the probability of homoploid hybrid speciation, i.e. the formation of a new species as a result of an hybridization event without changes in the ploidy of the organism, is a hotly debated topic. Here, we analyze a minimal model for homoploid hybrid speciation, in which reproductive isolation is achieved by means of (postzygotic) Dobzhansky-Muller incompatibilities. When these postzygotic genetic incompatibilities are resolved in the hybrid population, their reciprocal sorting can result in reproductive isolation from both parental populations, thus creating a hybrid species. We show that, in accordance with the current literature, hybrid speciation tends to be rare. However, specific arrangements of the genes responsible for reproductive isolation can make reciprocal sorting almost unavoidable and thus create barriers to the parental population in an almost deterministic matter. We discuss the implications of these results for hybrid speciation and patterns of introgression in nature.

International audience; Classical methods, used for large scale treatments such as mechanical or chemical extractions, affect the integrity of extracted cytosolic protein by releasing proteases contained in vacuoles. Our previous experiments on flow processes electroextraction on yeasts proved that pulsed electric field technology allows preserving the integrity of released cytosolic proteins, by not affecting vacuole membranes. Furthermore, large cell culture volumes are easily treated by the flow technology. Based on this previous knowledge, we developed a new protocol in order to electro-extract total cytoplasmic proteins from microalgae (Nannochloropsis salina, Chlorella vulgaris and Haematococcus pluvialis). Given that induction of electropermeabilization is under the control of target cell size, as the mean diameter for N. salina is only 2.5 μm, we used repetitive 2 ms long pulses of alternating polarities with stronger field strengths than previously described for yeasts. The electric treatment was followed by a 24 h incubation period in a salty buffer. The amount of total protein release was observed by a classical Bradford assay. A more accurate evaluation of protein release was obtained by SDS-PAGE. Similar results were obtained with C. vulgaris and H. pluvialis under milder electrical conditions as expected from their larger size.

The rise of human populations and the growth of cities contribute to the depletion of natural resources, increase their cost, and create potential climatic changes. To overcome difficulties in supplying populations and reducing the resource cost, a search for alternative pharmaceutical, nanotechnology, and energy sources has begun. Among the alternative sources, microalgae are the most promising because they use carbon dioxide (CO2) to produce biomass and/or valuable compounds. Once produced, the biomass is ordinarily harvested and processed (downstream program). Drying, grinding, and extraction steps are destructive to the microalgal biomass that then needs to be renewed. The extraction and purification processes generate organic wastes and require substantial energy inputs. Altogether, it is urgent to develop alternative downstream processes. Among the possibilities, milking invokes the concept that the extraction should not kill the algal cells. Therefore, it does not require growing the algae anew. In this review, we discuss research on milking of diatoms. The main themes are (a) development of alternative methods to extract and harvest high added value compounds; (b) design of photobioreactors; (c) biodiversity and (d) stress physiology, illustrated with original results dealing with oleaginous diatoms.

IntroductionMedical problems are frequently encountered during electronic dance music (EDM) events.ProblemThere are uncertainties about the frequencies and severity of intoxications with different types of recreational drugs: ethanol, “classical” illicit party drugs, and new psychoactive substances (NPS).MethodsStatistical data on the medical problems encountered during two editions of an indoor electronic dance event with around 30,000 attendants were retrieved from the Belgian Red Cross (Mechelen, Belgium) database. Data on drug use were prospectively collected from the patient (or a bystander), the clinical presentation, and/or toxicological screening.ResultsIn the on-site medical station, 487 patients were treated (265 in 2013 and 222 in 2014). The most frequent reasons were trauma (n=171), headache (n=36), gastro-intestinal problems (n=44), and intoxication (n=160). Sixty-nine patients were transferred to a hospital, including 53 with severe drug-related symptoms. Analysis of blood samples from 106 intoxicated patients detected ethanol in 91.5%, 3,4-methylenedioxymethamphetamine (MDMA) in 34.0%, cannabis in 30.2%, cocaine in 7.5%, amphetamine in 2.8%, and gamma-hydroxybutyric acid (GHB) in 0.9% of patients (alone or in combination). In only six of the MDMA-positive cases, MDMA was the sole substance found. In 2014, the neuroleptic drug clozapine was found in three cases and ketamine in one. Additional analyses for NPS were performed in 20 cases. Only in one agitated patient, the psychedelic phenethylamines 25B-NBOMe and 25C-NBOMe were found.ConclusionsAt this particular event, recreational drug abuse necessitated on-site medical treatment in one out of 350 attendants and a hospital transfer in one out of 1,000. Ethanol remains the most frequently abused (legal) drug, yet classical illicit recreational drugs are also frequently (co-) ingested. The most worrying observation was high-risk poly-drug use, especially among MDMA users. Regarding NPS, the number of cases was low and the clinical presentations were rather mild. It should be stressed that these observations only apply to this particular event and cannot be generalized to other EDM events.CalleP, SundahlN, MaudensK, WilleSMR, Van SassenbroeckD, De GraeveK, GogaertS, De PaepeP, DevrieseD, ArnoG, BlanckaertP. Medical emergencies related to ethanol and illicit drugs at an annual, nocturnal, indoor, electronic dance music event. Prehosp Disaster Med. 2018;33(1):71–76.

The deposited article version is a post-print and is the "Accepted Author Manuscript" provided by Wiley and posted online on 12th January 2018. This publication hasn't any creative commons license associated. This deposit is composed by the main article plus the supplementary materials of the publication. In many diploid species the sex chromosomes play a special role in mediating reproductive isolation. In haplodiploids, where females are diploid and males haploid, the whole genome behaves similarly to the X/Z chromosomes of diploids. Therefore, haplodiploid systems can serve as a model for the role of sex chromosomes in speciation and hybridization. A previously described population of Finnish Formica wood ants displays genome-wide signs of ploidally and sexually antagonistic selection resulting from hybridization. Here, hybrid females have increased survivorship but hybrid males are inviable. To understand how the unusual hybrid population may be maintained, we developed a mathematical model with hybrid incompatibility, female heterozygote advantage, recombination, and assortative mating. The rugged fitness landscape resulting from the co-occurrence of heterozygote advantage and hybrid incompatibility results in a sexual conflict in haplodiploids, which is caused by the ploidy difference. Thus, whereas heterozygote advantage always promotes long-term polymorphism in diploids, we find various outcomes in haplodiploids in which the population stabilizes either in favor of males, females, or via maximizing the number of introgressed individuals. We discuss these outcomes with respect to the potential long-term fate of the Finnish wood ant population, and provide approximations for the extension of the model to multiple incompatibilities. Moreover, we highlight the general implications of our results for speciation and hybridization in haplodiploids versus diploids, and how the described fitness relationships could contribute to the outstanding role of sex chromosomes as hotspots of sexual antagonism and genes involved in speciation. This article is protected by copyright. All rights reserved. Fundação Calouste Gulbenkian; National Science Foundation grant: (NSF PHY-1125915); Human Frontier Science Program; Finnish Cultural Foundation; Academy of Finland grant: (252411). info:eu-repo/semantics/acceptedVersion

Aims This article shows the effect of nanosecond pulsed electric field (nsPEF) on Escherichia coli, which could imply a durable change in protein expressions and then impacted the phenotype of surviving bacteria that might lead to increase pathogenicity. Methods and Results The effects of nsPEF on E. coli viability and membrane permeabilization were investigated. One log10 reduction in bacterial counts was achieved at field strength of 107 V m−1 with a train of 500 successive pulses of 60 × 10−9 s. Incubation of germs after treatment with propidium iodide showed that membrane permeabilization was reversible. Possible protein changes of surviving bacteria were checked to assess potential phenotypical changes using two-dimensional electrophoresis. In our study, after 40 generations, only UniProt #P39187 was up-regulated with P ≤ 0·05 compared with the control and corresponded to the uncharacterized protein YtfJ. Antibiograms were used to check whether or not the pattern of cultivable bacteria after nsPEF deliveries changed. Conclusions The results tend to show that nsPEFs are able to inactivate bacteria and have probably no serious impact in E. coli protein patterns. Significance and Impact of the Study The use of nsPEF is a safe promising new nonthermal method for bacterial inactivation in the food processing and environmental industry.

SALMO (Simulation by means of an Analytical Lake Model) describes the seasonal dynamics of PO 4 -P, NO 3 -N, detritus, chlorophyta, bacillariophyta, cyanophyta and cladocerans of the epilimnion and hypolimnion of stratified lakes by complex ordinary differential equations. This study adapted the model for shallow polymictic lakes and developed a real-coded genetic algorithm to optimize key parameters identified by sensitivity analysis. Meiliang Bay of Taihu Lake is taken as a study case, and 5 years data were collected for model calibration and validation. Given the good performance of the adapted and optimized SALMO, the model was applied to analyze four management scenarios for the Meiliang Bay, including 50% reduction of external nutrient loads, 50% reduction of internal nutrient loads, 50% reduction of zooplankton mortality and 1 °C increase of water temperature. The study showed: the adapted SALMO model is applicable to shallow polymictic and hypertrophic lakes; the real-coded genetic algorithm significantly improved the model efficiency and accuracy; reduction of nutrients from sediment release appears to be a key measure for controlling cyanobacteria blooms in Lake Taihu; biomanipulation is very delicate and can only be implemented with sufficient investigation and great caution; the increase of algae biomass due to water temperature rise indicates extra potential threat from climate change to lake ecosystems.

Mutation may impose a substantial load on populations, which varies according to the reproductive mode of organisms. Over the past years, various authors used adaptive landscape models to predict the long-term effect of mutation on mean fitness; however, many of these studies assumed very weak mutation rates, so that at most one mutation segregates in the population. In this article, we derive several simple approximations (confirmed by simulations) for the mutation load at high mutation rate (U), using a general model that allows us to play with the number of selected traits (n), the degree of pleiotropy of mutations, and the shape of the fitness function (which affects the average sign and magnitude of epistasis among mutations). When mutations have strong fitness effects, the equilibrium fitness inline image of sexuals and asexuals is close to inline image; under weaker mutational effects, sexuals reach a different regime where inline image is a simple function of U and of a parameter describing the shape of the fitness function. Contrarily to weak mutation results showing that inline image is an increasing function of population size and a decreasing function of n, these parameters may have opposite effects in sexual populations at high mutation rate.

The paper reports the development of an individual-based fish dynamics model, of which the key components are the rules for the movement of individual fish and the definition of the habitat suitability. The distribution of the fish mainly depends on the flow conditions (velocity, depth, substrate) and life cycle of the fish. A major contribution is the refinement of the rules for fish movement, based on laboratory experiments under volitional swimming conditions, which also provided the ranges of preferential velocities and substrate size for the target fish, Spinibarbus hollandi. Moreover, they provided data on the fish trajectories and distribution patterns that allowed for validation of the movement rules. The validated fish dynamics model was applied to investigate the effect of discharge increase during the dry season by means of reservoir operation in the Lijiang River, which was the subject of field investigations in 2007 and 2008. The model results indicated that reservoir operation leads to an increase of fish biomass. According to the fish movement rules, fish cannot always escape from riverbed regions that dry during decreasing discharge events, which causes them to be trapped and die. Reservoir operation decreases the area of dry riverbed and reduces the travel distance for fish to escape from dry regions. Critical advantages of the individual fish model over global models defined on the population level are that they can account for the time that the fish needs to reach a region of suitable habitat and for the spatial pattern of suitable zones and their connectivity. Copyright © 2013 John Wiley & Sons, Ltd.

Use and management of wild and weedy species may involve artificial selection, which can determine morphological, physiological, reproductive, and genetic divergences between wild and managed populations, resulting in the initial or incipient phases of plant domestication. In this study we combined ethnobotanical, morphological, phytochemical and genetic information for analyzing differences between managed and unmanaged populations of the Mexican edible weed, Epazote (Chenopodium ambrosioides L.), in Santa Maria Tecomavaca, Oaxaca, a rural community within the Tehuacan-Cuicatlan Biosphere Reserve, Mexico. Our hypothesis was that differences in morphology (e.g. leave dimensions and density of pubescence), phytochemistry (e.g. flavor, odor, and amount of strong scented and toxic compounds) and/or molecular genetic markers, between managed and wild populations of Epazote in Santa Maria Tecomavaca, would indicate that managed populations have been and/or are under a process of incipient domestication. Our results revealed the existence in the study area of morphological variants associated with a gradient of management intensity, which involved apparent improved palatability correlated with a lowering of chemical defense. Most remarkably, we found agreement in the groupings defined by the cluster analyses of morphological and genetic data. Although Epazote is considered a weed or, at best, a minor crop, the results from four lines of evidence (cultural differentiation patterns, gigantism, reduction in chemical defenses, toxic compounds and inheritance of adaptive traits) suggest the existence of an incipient domestication process in the study zone.

Background and purpose: Rhythmical transient constrictions of the lymphatic vessels provide the means for efficient lymph drainage and interstitial tissue fluid balance. This activity is critical during inflammation, to avoid or limit oedema resulting from increased vascular permeability, mediated by the release of various inflammatory mediators. In this study, we investigated the mechanisms by which prostaglandin E2 (PGE2) and prostacyclin modulate lymphatic contractility in isolated guinea pig mesenteric lymphatic vessels. Experimental approach: Quantitative RT-PCR was used to assess the expression of mRNA for enzymes and receptors involved in the production and action of PGE2 and prostacyclin in mesenteric collecting lymphatic vessels. Frequency and amplitude of lymphatic vessel constriction were measured in the presence of these prostaglandins and the role of their respective EP and IP receptors assessed. Key results: Prostaglandin E2 and prostacyclin decreased concentration-dependently the frequency, without affecting the amplitude, of lymphatic constriction. Data obtained in the presence of the EP4 receptor antagonists, GW627368x (1 µM) and AH23848B (30 µM) and the IP receptor antagonist CAY10441 (0.1 µM) suggest that PGE2 predominantly activates EP4, whereas prostacyclin mainly stimulates IP receptors. Inhibition of responses to either prostaglandin with H89 (10 µM) or glibenclamide (1 µM) suggested a role for the activation of protein kinase A and ATP-sensitive K+ channels. Conclusions and implications: Our findings characterized the inhibition of lymphatic pumping induced by PGE2 or prostacyclin in guinea pig mesenteric lymphatics. This action is likely to impair oedema resolution and to contribute to the pro-inflammatory actions of these prostaglandins.

Background: Serum samples from patients with autoimmune connective tissue diseases that show a finely speckled antinuclear antibody (ANA) on indirect immune-fluorescence often have antibodies against unknown nuclear target antigens. To search for such autoantigens we applied a proteomic approach using sera from patients with a high ANA titer (≥640) and finely speckled fluorescence but in whom no antibodies to extractable nuclear antigens (ENA) could be identified.Methods: Using an immunoproteomics approach we identified heterogeneous nuclear ribonucleoprotein H1 (hnRNP H1) as a novel nuclear target of autoantibody response.Results: Recombinant rat hnRNP H1 reacted in Western blot analyses with 48% of 93 sera from patients with primary Sjögren syndrome and with 5.2% of 153 sera from patients with other connective tissue diseases (diseased controls). For comparison, the diagnostic sensitivity and specificity of anti–Sjögren syndrome A (SSA) antibodies for primary Sjögren syndrome in the same patient cohort were 88.2% and 76.3%, respectively. Interestingly, 5 of 11 primary Sjögren syndrome patients with no anti-SSA or anti-SSB antibodies had anti–hnRNP H1 antibodies. Anti–hnRNP H1 antibodies were preabsorbed by hnRNP H1, as demonstrated by indirect immunofluorescence. In an evaluation of the presence of anti–hnRNP H1 antibodies in 188 consecutive samples submitted to the clinical laboratory with positive ANA (titer ≥160), anti–hnRNP H1 antibodies were found in 3 of 7 (2 primary and 5 secondary) Sjögren syndrome patients and in 8.3% of the diseased controls.Conclusions: HnRNP H1 is a newly discovered autoantigen that could become an additional diagnostic marker.

Background and aims: Treatment with carbon monoxide (CO) inhalation has been shown to ameliorate postoperative ileus (POI) in rodents and swine. The aim of this study was to investigate whether CO liberated from water-soluble CO-releasing molecules (CO-RMs) can protect against POI in mice and to elucidate the mechanisms involved. Methods: Ileus was induced by surgical manipulation of the small intestine (IM). Intestinal contractility–transit was evaluated by video-fluorescence imaging. Leucocyte infiltration (myeloperoxidase), inflammatory parameters (ELISA), oxidative stress (lipid peroxidation), and haem oxygenase (HO)/inducible nitric oxide synthase (iNOS) enzyme activity were measured in the intestinal mucosa and muscularis propria. Results: Intestinal contractility and transit were markedly restored when manipulated mice were pre-treated with CO-RMs. Intestinal leucocyte infiltration, expression levels of interleukin 6 (IL6), monocyte chemoattractant protein-1 and intercellular adhesion molecule-1, as well as iNOS activity were reduced by treatment with CORM-3 (a transition metal carbonyl that releases CO very rapidly); whereas expression of IL10/HO-1 was further increased when compared to nontreated manipulated mice. Moreover, treatment with CORM-3 markedly reduced oxidative stress and extracellular signal-related kinase (ERK)1/2 activation in both mucosa (early response) and muscularis (biphasic response). The p38 mitogen-activated protein kinase inhibitor SB203580 abolished CORM-3-mediated HO-1 induction. The HO inhibitor chromium mesoporphyrin only partially reversed the protective effects of CORM-3 on inflammation/oxidative stress in the muscularis, but completely abrogated CORM-3-mediated inhibition of the early “oxidative burst” in the mucosa. Conclusions: Pre-treatment with CO-RMs markedly reduced IM-induced intestinal muscularis inflammation. These protective effects are, at least in part, mediated through induction of HO-1, in a p38-dependent manner, as well as reduction of ERK1/2 activation. In addition, CORM-induced HO-1 induction reduces the early “oxidative burst” in the mucosa following IM.

This study introduces a novel, simplified method for the evaluation of murine intestinal transit and contractility using fluorescence and video imaging. Intestinal transit was measured by evaluating the intestinal distribution of non-absorbable fluorescein-labelled dextran (70 kDa, FD70) along the gastrointestinal (GI) tract. After excision of the GI tract, two full-field images – one in normal illumination mode and another in fluorescent mode – were taken with a charge coupled device (CCD) camera and subsequently matched for calculation of fluorescence distribution along the GI tract. Immediately after, intestinal contractility was evaluated in different regions of the intact intestine by spatiotemporal motility mapping (i.e. video imaging). In control mice, the small intestine showed vigorous oscillatory contractions and FD70 was primarily distributed within the terminal ileum/caecum at 90 min postgavage. As validation step, the effect of intestinal manipulation (IM, surgical procedure) and two pharmacological agents – known to alter GI motility – was tested. At 24 h postoperatively, spontaneous contractile activity of the small intestine was nearly abolished in IM mice, leaving the small intestine distended and resulting in a significantly delayed intestinal transit. In accordance, spontaneous mechanical activity of circular muscle strips in standard organ baths was significantly reduced in IM mice compared to control mice. Administration of atropine (1–3 mg kg−1, i.p.) suppressed spontaneous contractile activity along the entire intestinal tract and induced a dose-related delay in intestinal transit. In contrast, metoclopramide (3–10 mg kg−1, i.p.) markedly increased contractile activity – however only in the upper GI tract – and accelerated intestinal transit in a dose-dependent manner.

Resume Objectifs Description d’epidemies recentes a Clostridium difficile. C. difficile est un bacille a Gram positif anaerobie sporule responsable de 15 a 25 % des cas de diarrhees post-antibiotiques et de plus de 95 % des cas de colites pseudomembraneuses (CPM). Depuis 2003, des epidemies de formes severes d’infections a C. difficile (ICD) ont ete decrites au Canada et aux Etats-Unis. Epidemiologie : particularites du clone 027 Ces epidemies sont liees a l’emergence et a la dissemination d’un clone particulier de C. difficile appele 027 (en reference a son profil en PCR-ribotypage) ou NAP1 (en reference a son profil en electrophorese en champ pulse). Ce clone a diffuse en Grande-Bretagne, en Belgique, aux Pays-Bas et plus recemment en France ou il a ete responsable d’epidemies, principalement dans la region Nord-Pas de Calais. Prevention Cette evolution preoccupante necessite un renforcement de la vigilance dans les etablissements de sante afin de controler la diffusion de ce clone.

International audience; Docosahexaenoic acid (DHA), the main member of the omega-3 essential fatty acid family, has been shown to reduce the invasion of the triple‑negative breast cancer cell line MDA‑MB‑231, but the mechanism involved remains unclear. In the present study, a proteomic approach was used to define changes in protein expression induced by DHA. Proteins from crude membrane preparations of MDA‑MB‑231 cells treated with 100 µM DHA were separated by two‑dimensional electrophoresis (2‑DE) and differentially expressed proteins were identified using MALDI‑TOF mass spectrometry. The main changes observed were the upregulation of Keratin, type Ⅱ cytoskeletal 1 (KRT1), catalase and lamin‑A/C. Immunocytochemistry analyses confirmed the increase in KRT1 induced by DHA. Furthermore, in vitro invasion assays showed that siRNA against KRT1 was able to reverse the DHA‑induced inhibition of breast cancer cell invasion. In conclusion, KRT1 is involved in the anti‑invasive activity of DHA in breast cancer cells.

VEB-1 extended-spectrum beta-lactamase-producing Acinetobacter baumannii was responsible for an outbreak in hospitals in France. A national alert was triggered in September 2003 when 4 hospitals reported clusters of A. baumannii infection with similar susceptibility profiles. Case definitions and laboratory guidelines were disseminated, and prospective surveillance was implemented; strains were sent to a single laboratory for characterization and typing. From April 2003 through June 2004, 53 hospitals reported 290 cases of A. baumannii infection or colonization; 275 isolates were bla(VEB-1)-positive and clonally related. Cases were first reported in 5 districts of northern France, then in 10 other districts in 4 regions. Within a region, interhospital spread was associated with patient transfer. In northern France, investigation and control measures led to a reduction of reported cases after January 2004. The national alert enabled early control of new clusters, demonstrating the usefulness of early warning about antimicrobial drug resist.

Summary A nosocomial outbreak of epidemiologically related VEB-1 extended-spectrum β-lactamase-producing isolates of Acinetobacter baumannii occurred in 33 patients in an intensive care unit. A case-control study identified previous treatment with third-generation cephalosporins as the only risk factor for A. baumannii acquisition. Rationale for antibiotic use should be strengthened.

Structural point mutations in exon 1 at codons 52, 54 and 57 and a promotor polymorphism at -221 bp of the mannose-binding lectin (MBL) gene are associated with increased susceptibility to various infectious diseases. We developed a genotyping method based on the 5' nuclease (TaqMan) assay in combination with the use of minor-groove-binder (MGB) probes in screening for these mutations/polymorphisms. In contrast to conventional probes, MGB probes have a short length and can be used for detection of mutations that are in close proximity to each other, as is the case for the structural mutations in exon 1 of the MBL gene. Results obtained with the 5' nuclease assay using MGB probes were identical with results obtained with classical techniques such as restriction fragment length polymorphism, allele-specific PCR, and sequencing. In conclusion, the 5' nuclease assay using MGB probes is useful for large-scale screening of point mutations/polymorphisms, even when they are in close proximity.

Protection against infections with Streptococcus pneumoniae is mediated by antibodies against the capsular polysaccharides (caps-PS). Here we show that in in vitro experiments CD4+ T lymphocytes stimulate and CD8+ T lymphocytes inhibit the human anti-caps-PS antibody response. Using antagonistic anti-CD40 and antagonistic anti-CD40 ligand (CD40L) monoclonal antibodies, we showed that the CD4+ T lymphocyte-mediated stimulation is dependent on the CD40-CD40L interaction. The role of CD40L was further illustrated by the observation that CD4+ T lymphocytes obtained from a patient with hyper-IgM syndrome were unable to enhance the immune response to caps-PS. Furthermore, CD4+ T lymphocytes from cord blood, which did not express CD40L in response to stimulation with caps-PS, failed to stimulate the antibody response of adult B lymphocytes to caps-PS. These in vitro findings were confirmed by in vivo experiments in which SCID/SCID mice were reconstituted with human mononuclear cells. Furthermore, we showed that caps-PS induce production of IL-4, IL-6, IL-10, and IFN-gamma, and that this enhanced production was inhibited by blocking the CD40-CD40L interaction. This is the first demonstration that the human immune response to caps-PS, which is markedly regulated by T lymphocytes, is dependent on the CD40-CD40L interaction.

Direct somatic embryogenesis was induced in leaf fragments of the Cichorium ‘474’ genotype. Addition of glycerol to the induction medium allowed a relative synchronization of the first division of the embryogenic cells that only occurs after transfer at day 5 to a medium without glycerol. The abundant presence of 9-kDa extracellular proteins in the culture-medium conditioned by somatic embryogenesis is reported here. Such proteins were also secreted when embryogenesis was initiated in root but were never detected when a non-embryogenic genotype was used as control under the same conditions of culture. Among these proteins, one basic and one acidic isoform were separated through cation-exchange chromatography. Both proteins were recognized by an antiserum raised against the carrot EP2 non-specific lipid transfer protein (nsLTP). In addition, the partial N-terminal amino acid sequence of each isoform showed similarities with nsLTPs of different plant species. The presence of the acidic nsLTP-like protein was concomitant with the obtention of embryogenic cells during the induction step. The basic form was shown to have only accumulated during the expression step when first divisions of embryogenic cells have occurred. These results allowed us to report, for the first time, the secretion of a 9-kDa acidic nsLTP-like protein in the culture-medium conditioned by plant embryogenic cells.

The aim of the present study is to evaluate in an elderly hospitalized population the diagnostic value of the serum transferrin receptor (sTfR) in distinguishing IDA (iron deficiency anemia) from ACD (anemia of chronic disease) as compared to conventional laboratory tests of iron metabolism, especially serum ferritin. In a prospective study, 34 patients with IDA and 38 patients with ACD (a chronic disorder in 23 and an acute infection in 15) were evaluated using iron status tests including serum transferrin receptor assay. The iron stores were assessed by bone marrow examination. sTfR levels were elevated (>28.1 nmol/L) in 68% of the IDA patients but also in 43% of the patients with ACD-chronic inflammation and 33% with ACD-acute infection. Serum ferritin was the best test to differentiate IDA from ACD patients. We conclude that serum ferritin is a more sensitive and specific parameter than the sTfR assay to predict the bone marrow iron status in an elderly anemic population.

Quantitative ultrasound (QUS) of bone is a valuable tool in the assessment of postmenopausal osteoporosis. QUS and new markers of bone turnover have been poorly assessed in Cushing's syndrome, however. Twenty-five patients with Cushing's syndrome (20 women, 3 men; mean age +/- SEM: 38+/-2 years) were studied and compared with 35 age- and sex-matched control patients (mean age +/- SEM: 38+/-2 years). The following variables were measured in both groups: QUS parameters at the heel (BUA; SOS; Stiffness Index, SI); bone mineral density (BMD) at both the lumbar spine (LS) and femoral neck (FN) by dual-energy X-ray absorptiometry; and serum markers of bone turnover (osteocalcin, procollagen type I N- and C-terminal propeptides (PINP and PICP), bone alkaline phosphatase (BAP), procollagen type I C-terminal telopeptide (ICTP) and urinary type I collagen C-telopepetide breakdown products (CTX)). Both BUA and SI were decreased in patients with Cushing's syndrome (p0.01) but not SOS (p=0.08). BMD was also strongly decreased in Cushing's syndrome, at both the LS and FN (p0.005). The two markers of bone turnover statistically significantly different between the two groups were osteocalcin (mean + SEM: 3.5 + 0.7 ng/ml (Cushing's syndrome) vs 6.4+/-0.5 ng/ml (controls, p0.01)) and CTX (mean +/- SEM: 148.7+/-17.1 microg/mmol Cr (Cushing's syndrome) vs 220.8+/-22.9 microg/mmol Cr (controls), p0.05). The areas under the receiver operating characteristic curve (AUC) were 0.72 (BUA), 0.73 (SI), 0.90 (BMD(LS)), 0.81 (BMD(FN)), 0.83 (osteocalcin) and 0.64 (CTX) respectively. AUC was significantly higher for BMD(LS) than for both BUA and SI (p0.05). Conversely AUC was not statistically significantly different for BMDFN as compared with either BUA or SI. AUC was also higher for osteocalcin than for other markers of bone turnover. In conclusion, QUS of bone seems to be a relevant tool for assessing bone involvement in Cushing's syndrome. QUS does have a lower sensitivity compared with DXA, however, and the relevance of QUS cannot be ascertained until some longitudinal data are forthcoming. Except for CTX, the other new markers of bone turnover assessed in this study (PINP, PICP, BAP and ICTP) do not seem of interest in Cushing's syndrome.

Quantitative ultrasound (QUS) of bone and new markers of bone remodeling have been poorly investigated in mild primary hyperparathyroidism (PHPT). In this study 26 patients (20 females and 6 males) were evaluated. BUA and SOS were measured by QUS at the heel. Markers of bone remodeling assessed were bone alkaline phosphatase (BAP), osteocalcin (OC), procollagen type I N- and C-terminal propeptides (PINP et PICP), and procollagen type I C-terminal telopeptide in blood and urine (ICTP and CTX). Bone mineral density (BMD) was measured at the lumbar spine (LS), femoral neck (FN), and Ward's triangle (WT). The control group comprised 35 sex- and age-matched subjects. The statistically significant variables between the two groups were (P0.05) BUA, BMD(LS), BMD(FN), BMD(WT), BAP, and OC. Corresponding z-scores were -0.55 +/- 0.75, -0.66 +/- 0.77, -0.66 +/- 0.71, -0.67 +/- 0.52, 1.87 +/- 3.87, and 1.93 +/- 3.53, respectively. Although PICP and PINP levels were higher in PHPT patients as compared with controls, the difference was not significant. Several markers of bone turnover were moderately correlated with both QUS (r = -0.39 to -0.55) and BMD (r = -0.48 to 0.63). In conclusion QUS seems to be a relevant tool in the assessment of bone status for patients with mild PHPT.

Anti-cyclic citrullinated peptide (CCP) antibodies and rheumatoid factor (RF) are recommended screening tests for rheumatoid arthritis.1 In a meta-analysis,2 the sensitivity of anti-CCP (67%) for diagnosing rheumatoid arthritis was comparable to the sensitivity of RF (69%).2 The specificity of anti-CCP for rheumatoid arthritis (95%) was higher than the specificity of RF (85%).2 Consequently, the positive likelihood ratio was higher for anti-CCP (12.46) than for RF (4.86).2 Studies that addressed the clinical usefulness of anti-CCP used a single cut-off value and, hence, likelihood ratios were calculated based on a single cut-off. In the present letter, we illustrate how likelihood ratios for anti-CCP and RF …

Cardiac troponin I assays for Axsym (Abbott Diagnostics, Abbott Park, IL, USA) and Immuno 1 (Bayer Corporation, Tarrytown, NY, USA) analysers were evaluated. Heparin plasma or serum could be used for both assays. Samples were stable for 24 h at ambient temperature, 3 days at 4-8 degrees C and 3 months at -20 degrees C. After 10 months' storage at -80 degrees C, the recoveries were well above 100% by both assays. Total coefficients of variation for Axsym assay were 9.0%, 5.8% and 5.3% at concentrations of 2.6 microg/l, 9.83 microg/l and 34.3 microg/l respectively; for Immuno 1 these were 4.4 %, 1.6% and 1.8% at 2.3 microg/l, 6.27 microg/l and 44.35 microg/l respectively. It was > or =20% at concentration of < or =0.5 microg/l for Axysm assay and < or =0.15 microg/l for Immuno 1 assay. Recoveries were < or =90% at < or =0.22 microg/l on Axsym and at < or =1.47 microg/l on Immuno 1. Neither method showed significant interference with haemoglobin, bilirubin, triglycerides or rheumatoid factor. Correlation between the two methods was excellent (r = 0.997, Y (Axsym) = 4.2X (Immuno 1) +3.2). The highest concentrations detected in 50 healthy subjects were 0.3 microg/l and 0.1 microg/l by Axsym and Immuno 1 methods, respectively. Twelve out of 43 renal failure patients had troponin I 0.13-0.9 microg/l using Axsym method and 4 had levels of 0.07-0.13 microg/l using Immuno 1. In muscle trauma patients, troponin I was undetectable.

We compared the automated Paragon 2000 clinical capillary zone electrophoresis (CZE) system with two manual methods, agarose electrophoresis (AGE) and cellulose acetate electrophoresis (CAE). Reference intervals in healthy adults were determined for each method. When compared with AGE and CAE, CZE gave substantially higher reference values for the α1-globulin fraction. With CZE, within-run precision for fraction quantitation was between 0.5% (albumin) and 4.1% (α1-globulin). Total precision was between 0.8% (albumin) and 5.3% (β-globulin). Data obtained from CZE showed poor linear correlation with results obtained by AGE but good linear correlation with data from CAE. Analysis of serum from patients with inter alia inflammation, nephrotic syndrome, or polyclonal gammopathy showed that clinical information obtained by CZE is comparable with information obtained by AGE and CAE. We conclude that CZE offers a clinically reliable alternative to AGE and CAE and has the advantages of automation, higher precision, and faster turnaround time.

A selection of 58 specimens with a monoclonal component identified by immunoelectrophoresis and/or immunofixation was analyzed with the immunosubtraction procedure on the Paragon 2000 capillary electrophoresis system. The capillary system detected 93% of the paraproteins and, using immunosubtraction, correctly identified 91% of the paraproteins. Paraproteins that were detected by immunofixation and/or immunoelectrophoresis but not by capillary electrophoresis were also missed by agarose electrophoresis and cellulose acetate electrophoresis. Cellulose acetate electrophoresis was the least sensitive method for detection of paraproteins. Only 74% of the monoclonal components were detected by this technique, whereas 86% were revealed by agarose electrophoresis. In addition to monoclonal paraproteins, we also studied biclonal paraproteins and oligoclonal banding. Capillary electrophoresis and immunosubtraction correctly detected and identified three specimens containing biclonal paraproteins. In one specimen, capillary zone electrophoresis detected only one band, whereas agarose gel electrophoresis detected two bands. The sensitivity for detection and identification of oligoclonal banding by capillary electrophoresis was inferior to immunofixation.

The quantification of monoclonal immunoglobulins by protein electrophoresis has been helpful in determining the prognosis for the patient. In a 65-year old man with documented IgG lambda monoclonal gammopathy, a discrepancy in the detection and quantification of this M-protein was found when studied by four different electrophoretic methods. On a high-resolution agarose method with acid violet staining a prominent peak was seen in the fast gamma-region. A lower resolution agarose method using an amido black staining showed a small mid-gamma restriction, and no spike was detected by high-resolution or lower resolution capillary zone electrophoresis. The discrepancy was not attributable to a migration in the beta-region leading to masking by transferrin or C3 peak, incorrect separation on CZE due to a high isoelectric point (pI > 8.5) or pentamerization of IgM M-proteins. Therefore, other causes for discrepancies should be investigated.

Capillary zone electrophoresis (CZE) is documented to have high sensitivity and specificity for the detection of monoclonal (M) proteins. In two large (>1500 participants) prospective studies in which CZE was compared with “immunofixation/immunoelectrophoresis”, the sensitivity of CZE was 95% (1)(2). This is superior to the sensitivity of agarose gel electrophoresis (AGE), which Katzmann et al. (1) found to be 91% and Meunier (3) found to be 92.2% (evaluation of 2060 sera containing an M-protein). Especially in the β-globulin region, CZE with Paragon CZE 2000 (Beckman-Coulter) is superior to AGE for detection of low-concentration M-proteins (IgA and light chains) (3)(4). The specificity of CZE is reported to be high (98.6%) and comparable to that of AGE (1)(5). In a prospective study (2), we described three M-proteins with high pI values that migrated in the slow γ region on AGE but were not detected with Paragon. In addition, one high-concentration IgG|gl with a pI ≈7 that migrated in the mid-γ region on AGE was not separated by CZE. The Paragon has recently been upgraded [modified buffer, higher voltage (10.3 kV), more efficient cooling, adapted software (1.6.02)], and according … [↵][1]bAuthor for correspondence. [1]: #xref-corresp-2-1

Spores are dormant units of bacteria resistant to numerous disinfection methods. Additionally, the effects on bacteria of repetitive electrical discharges in water by used of the so-called "corona discharges" or streamer are poorly described. In this study vegetative and spore forms of Bacillus subtilis var. niger were subjected to these discharges. To generate corona discharges in water, a Marx generator capable of delivering 60-90 kV was used with a coaxial chamber of treatment. Vegetative and spore form reductions were defined using colony-forming unit counting. Proteins extracts were separated by two-dimensional electrophoresis and spots of interest were characterized by mass spectrometry. Shock waves were assessed by the diminution of liposome size and OD(400 nm). The results show a decrease in bacteria viability of 2 log(10) after 1000 discharges on the vegetative form and 4 log(10) after 10,000 discharges on the spores. Two-dimensional electrophoresis showed that the streamers impact the regulation of several proteins in the vegetative forms with UniProt ID: P80861, Q06797, P80244, C0ZI91, respectively. The reduction appears to be due, in part, to hydrogen peroxide (H(2)O(2)) generated by the corona discharges while spore deactivation remained insensitive to these chemicals. The spore eradication was associated to shock waves induced by the discharges but not H(2)O(2). Corona discharges appear as a prospective method for eradication of spores in water. The corona discharges can be an efficient method for decontamination processes of waste water.

International audience