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A new marine benthic toxic Prorocentrum species is described from the tropical/subtropical regions of the Atlantic (Colombian Caribbean Sea and Northeast Brazil) and Pacific (Southern Japan) oceans. Morphological cell structures were examined using light (LM) and scanning electron (SEM) microscopy. Prorocentrum porosum sp. nov. was characterized by 35.9-50.2 μm long and 25.4-45.7 μm deep cells, covered by broadly ovoid symmetric thecal plates. The surface of both thecal plates is smooth and covered by randomly scattered kidney-shaped pores (n = 102-149), rounder towards the center, absent in the central part, and surrounded by a conspicuous marginal ring of about 69-92 evenly spaced pores. Broad V-shaped periflagellar area exhibiting flagellar and accessory pores. The molecular phylogenetic position of P. porosum sp. nov. was inferred using partial LSU rRNA gene (rDNA) and rDNA ITS sequences. This new species branched with high support in a Prorocentrum clade including P. caipirignum, P. hoffmannianum and P. cf. lima (P. lima morphotype 5 sensuZhang et al., 2015). Pairwise comparison of ITS1 and ITS2 transcripts with these closest relatives revealed the presence of compensatory base changes (CBCs), with the exception of P. cf. lima (P. lima morphotype 5), which only showed in ITS2 a hemi-CBC (HCBC) and two base changes that possibly induce a structural modification. Toxin analyses performed in two Colombian and Brazilian strains in the present study detected the presence of low amounts of okadaic acid., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022. Published by Elsevier B.V.)

ABC ("ATP-Binding Cassette") transporters of the type IV subfamily consist of exporters involved in the efflux of many compounds, notably those capable to confer multidrug resistance like the mammalian P-glycoprotein or the bacterial transporter BmrA. They function according to an alternating access mechanism between inward-facing (IF) and outward-facing (OF) conformations, but the extent of physical separation between the two nucleotide-binding domains (NBDs) in different states is still unsettled. Small Angle Neutron Scattering and hydrogen/deuterium exchange coupled to mass spectrometry were used to highlight different conformational states of BmrA during its ATPase cycle. In particular, mutation of the conserved Lysine residue of the Walker-A motif (K380A) captures BmrA in an ATP-bound IF conformation prior to NBD closure. While in the transition-like state induced by vanadate wild-type BmrA is mainly in an OF conformation, the transporter populates only IF conformations in either the apo state or in the presence of ADP/Mg. Importantly, in this post-hydrolytic step, distances between the two NBDs of BmrA seem to be more separated than in the apo state, but they remain shorter than the widest opening found in the related MsbA transporter. Overall, our results highlight the main steps of the catalytic cycle of a homodimeric bacterial multidrug transporter and underline structural and functional commonalities as well as oddities among the type IV subfamily of ABC transporters., Competing Interests: Declaration of interests The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

During infections, fast identification of the microorganisms is critical to improve patient treatment and to better manage antibiotics use. Electrochemistry exhibits several advantages for rapid diagnostic: it enables easy, cheap and in situ analysis of redox molecules in most liquids. In this work, several culture supernatants of different Pseudomonas aeruginosa strains (including PAO1 and its isogenic mutants PAO1ΔpqsA, PA14, PAK and CHA) were analyzed by square wave voltammetry on glassy carbon electrode during the bacterial growth. The obtained voltamograms shown complex traces exhibiting numerous redox peaks with potential repartitions and current amplitudes depending on the studied bacterium and/or growth time. Among them, some peaks were clearly associated to the well-known redox toxin Pyocyanin (PYO) and the autoinducer Pseudomonas Quinolone Signal (PQS). Other peaks were observed that are not yet attributed to known secreted species. Each complex electrochemical response (number of peaks, peak potential and amplitude) can be interpreted as a fingerprint or "ID-card" of the studied strain that may be implemented for fast bacteria strain identification., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Published by Elsevier B.V.)

TiO 2 particles are widely used in products for everyday consumption, such as cosmetics and food; their possible adverse effects on human health must therefore be investigated. The aim of this study was to document in vitro impact of the food additive E171, i.e. TiO 2 , and of TiO 2 nanoparticles, on a co-culture of Caco-2 and HT29-MTX cells, which is an in vitro model for human intestine. Cells were exposed to TiO 2 particles three days after seeding, i.e. while they were not fully differentiated. Cell viability, reactive oxygen species (ROS) levels and DNA integrity were assessed, by MTT assay, DCFH-DA assay, alkaline and Fpg-modified comet assay and 8-oxo-dGuo measurement by HPLC-MS/MS. The mRNA expression of genes involved in ROS regulation, DNA repair via base-excision repair, and endoplasmic reticulum stress was assessed by RT-qPCR. Exposure to TiO 2 particles resulted in increased intracellular ROS levels, but did not impair cell viability and did not cause any oxidative damage to DNA. Only minor changes in mRNA expression were detected. Altogether, this shows that E171 food additive and TiO 2 nanoparticles only produce minor effects to this in vitro intestinal cell model., (Copyright © 2018 Elsevier B.V. All rights reserved.)

In natural variable environments, plants rapidly adjust photosynthesis for optimum balance between photochemistry and photoprotection. These adjustments mainly occur via changes in their proton motive force (pmf). Recent studies based on time resolved analysis of the Electro Chromic Signal (ECS) bandshift of photosynthetic pigments in the model plant Arabidopsis thaliana have suggested an active role of ion fluxes across the thylakoid membranes in the regulation of the pmf. Among the different channels and transporters possibly involved in this phenomenon, we previously identified the TPK3 potassium channel. Plants silenced for TPK3 expression displayed light stress signatures, with reduced Non Photochemical Quenching (NPQ) capacity and sustained anthocyanin accumulation, even at moderate intensities. In this work we re-examined the role of this protein in pmf regulation, starting from the observation that both TPK3 knock-down (TPK3 KD) and WT plants display enhanced anthocyanin accumulation in the light under certain growth conditions, especially in old leaves. We thus compared the pmf features of young "green" (without anthocyanins) and old "red" (with anthocyanins) leaves in both genotypes using a global fit analysis of the ECS. We found that the differences in the ECS profile measured between the two genotypes reflect not only differences in TPK3 expression level, but also a modified photosynthetic activity of stressed red leaves, which are present in a larger amounts in the TPK3 KD plants., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Water scarcity is increasingly raising the need for non-conventional water resources, particularly in arid and semi-arid regions. In this context, atmospheric moisture can potentially be harvested in the form of dew, which is commonly disregarded from the water budget, although its impact may be significant when compared to rainfall during the dry season. In this study, a dew atlas for the Mediterranean region is presented illustrating dew yields using the yield data collected for the 2013 dry season. The results indicate that cumulative monthly dew yield in the region can exceed 2.8mm at the end of the dry season and 1.5mm during the driest months, compared to <1mm of rainfall during the same period in some areas. Dew yields were compared with potential evapotranspiration (PET) and actual evapotranspiration (ET) during summer months thus highlighting the role of dew to many native plants in the region. Furthermore, forecasted trends in temperature and relative humidity were used to estimate dew yields under future climatic scenarios. The results showed a 27% decline in dew yield during the critical summer months at the end of the century (2080)., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Microorganisms, such as bacteria, are one of the first targets of nanoparticles in the environment. In this study, we tested the effect of two nanoparticles, ZnO and TiO2, with the salt ZnSO4 as the control, on the Gram-positive bacterium Bacillus subtilis by 2D gel electrophoresis-based proteomics. Despite a significant effect on viability (LD50), TiO2 NPs had no detectable effect on the proteomic pattern, while ZnO NPs and ZnSO4 significantly modified B. subtilis metabolism. These results allowed us to conclude that the effects of ZnO observed in this work were mainly attributable to Zn dissolution in the culture media. Proteomic analysis highlighted twelve modulated proteins related to central metabolism: MetE and MccB (cysteine metabolism), OdhA, AspB, IolD, AnsB, PdhB and YtsJ (Krebs cycle) and XylA, YqjI, Drm and Tal (pentose phosphate pathway). Biochemical assays, such as free sulfhydryl, CoA-SH and malate dehydrogenase assays corroborated the observed central metabolism reorientation and showed that Zn stress induced oxidative stress, probably as a consequence of thiol chelation stress by Zn ions. The other patterns affected by ZnO and ZnSO4 were the stringent response and the general stress response. Nine proteins involved in or controlled by the stringent response showed a modified expression profile in the presence of ZnO NPs or ZnSO4: YwaC, SigH, YtxH, YtzB, TufA, RplJ, RpsB, PdhB and Mbl. An increase in the ppGpp concentration confirmed the involvement of the stringent response during a Zn stress. All these metabolic reorientations in response to Zn stress were probably the result of complex regulatory mechanisms including at least the stringent response via YwaC., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Although the biological effects of titanium dioxide nanoparticles (TiO2-NPs) have been studied for more than two decades, the mechanisms governing their toxicity are still unclear. We applied 2D-gel proteomics analysis on A549 epithelial alveolar cells chronically exposed for 2months to 2.5 or 50μg/mL of deeply characterized TiO2-NPs, in order to obtain comprehensive molecular responses that may reflect functional outcomes. We show that exposure to TiO2-NPs impacts the abundance of 30 protein species, corresponding to 22 gene products. These proteins are involved in glucose metabolism, trafficking, gene expression, mitochondrial function, proteasome activity and DNA damage response. Besides, our results suggest that p53 pathway is activated, slowing down cell cycle progression and reducing cell proliferation rate. Moreover, we report increased content of chaperones-related proteins, which suggests homeostasis re-establishment. Finally, our results highlight that chronic exposure to TiO2-NPs affects the same cellular functions as acute exposure to TiO2-NPs, although lower exposure concentrations and longer exposure times induce more intense cellular response., Biological Significance: Our results make possible the identification of new mechanisms that explain TiO2-NP toxicity upon long-term, in vitro exposure of A549 cells. It is the first article describing -omics results obtained with this experimental strategy. We show that this long-term exposure modifies the cellular content of proteins involved in functions including mitochondrial activity, intra- and extracellular trafficking, proteasome activity, glucose metabolism, and gene expression. Moreover we observe modification of content of proteins that activate the p53 pathway, which suggest the induction of a DNA damage response. Technically, our results show that exposure of A549 cells to a high concentration of TiO2-NPs leads to the identification of modulations of the same functional categories than exposure to low, more realistic concentrations. Still the intensity differs between these two exposure scenarios. We also show that chronic exposure to TiO2-NPs induces the modulation of cellular functions that have already been reported in the literature as being impacted in acute exposure scenarios. This proves that the exposure protocol in in vitro experiments related to nanoparticle toxicology might be cautiously chosen since inappropriate scenario may lead to inappropriate and/or incomplete conclusions., (Copyright © 2015 Elsevier B.V. All rights reserved.)

The cellular responses of the J774 macrophage cell line to zinc oxide and zirconium oxide nanoparticles have been studied by a comparative quantitative, protein level based proteomic approach. The most prominent results have been validated by targeted approaches. These approaches have been carried out under culture conditions that stimulate mildly the aryl hydrocarbon receptor, thereby mimicking conditions that can be encountered in vivo in complex environments. The comparative approach with two nanoparticles allows to separate the common responses, which can be attributed to the phagocytosis event per se, from the response specific to each type of nanoparticles. The zinc-specific responses are the most prominent ones and include mitochondrial proteins too, but also signaling molecules such as MyD88, proteins associated with methylglyoxal detoxification (glyoxalase 2, aldose reductase) and deoxyribonucleotide hydrolases. The in cellulo inhibition of GAPDH by zinc was also documented, representing a possible source of methylglyoxal in the cells, leading to an increase in methylglyoxal-modified DNA bases. These observations may be mechanistically associated with the genotoxic effect of zinc and its selective effects on cancer cells., Biological Significance: The responses of the murine J774 macrophage cell lines to two types of metallic oxide nanoparticles (zinc oxide and zirconium dioxide) were studied by a comparative 2D gel based approach. This allows sorting of shared responses from nanoparticle-specific responses. Zinc oxide nanoparticles induced specifically a strong decrease in the mitochondrial function, in phagocytosis and also an increase in the methylglyoxal-associated DNA damage, which may explain the well known genotoxicity of zinc. In conclusion, this study allows highlighting of pathways that may play an important role in the toxicity of the zinc oxide nanoparticles., (Copyright © 2015 Elsevier B.V. All rights reserved.)

The phosphorylation of the H2AX histone to form γH2AX foci has been shown to be an accurate biomarker of ionizing radiation exposure. It is well established that there is a one-to-one correlation between the number of γH2AX foci and radiation-induced double strand breaks in cellular DNA, which can be translated to the received dose. However, manual counting of foci is time-consuming, and cannot accommodate high throughput analysis required to obtain rapid results for medical triage purposes in the case of large-scale accidental exposure. Furthermore, the accuracy of γH2AX measurements could potentially be compromised by delays between the time of exposure and analysis of results, as well as inter-cellular and inter-individual variability of this biological response. To evaluate more rapid approaches of quantifying γH2AX for use in an emergency situation, and to determine the impact of inter-individual variability, we compared two methods of global γH2AX fluorescence quantification (low magnification immunofluorescence microscopy and flow cytometry) to the well-established γH2AX foci scoring method in human primary fibroblasts. All three approaches were well correlated, indicating that global γH2AX fluorescence measurements are suitable for dose estimation. For rapid triage in an emergency situation, we propose the use of flow cytometry, as it is more highly correlated with foci scoring and because of the speed and ease of the method. Dose response curves (0.25-6Gy) using flow cytometry measurements showed that inter-individual variability in global γH2AX fluorescence is statistically insignificant at 4h post-irradiation. Based on these data, we propose calibration curves that can be applied to populations exposed to moderate radiation doses to estimate individual received doses, independent of individual radiosensitivity, at this specific time point post-irradiation using human fibroblasts and lymphocytes. Furthermore, we define three triage categories that could facilitate immediate and follow-up care in the case of a radiological accident., (Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.)

We report on a female fetus with macrocephaly and macroglossia harbouring 13q31.1 microdeletion encompassing three genes: SPRY2, NDFIP2 and RBM26. NDFIP2 protein is involved in ubiquitination and in Ras/mitogen-activated protein kinase (MAPK) signaling pathways. SPRY2 protein is part of Sprout protein family and inhibits the Ras/MAPK pathways. Ras/MAPK pathway plays important role in complex cellular programs including cell differentiation and proliferation. Germline mutations in genes encoding protein involved in the MAPK cascade is responsible for a wide family of developmental disorders known as RASopathies. Some RASopathies, such as Costello syndrome, present a phenotype with (relative) macrocephaly as perinatal features. However, prenatal-onset macroglossia are generally absent in this syndrome but rather suggestive of the Beckwith-Wiedemann syndrome for which molecular testing were negative. Phenotype-genotype correlation with patients from DECIPHER defines NDFIP2 and SPRY2 as a possible candidate genes for a RASopathy potentially responsible for the clinical features in the fetus. Finally, this original case of 13q31.1 microdeletion underlines the importance of array-CGH in prenatal diagnosis with sonographic signs such as macroglossia and/or macrocephaly. In this case, genetic investigation should be not limited to the search of well-known genetic causes and other genomic microdeletions should be considered as alternative diagnoses for macroglossia., (Copyright © 2015 Elsevier Masson SAS. All rights reserved.)

The unique physicochemical properties of nanomaterials has led to an increased use in the paint and coating industry. In this study, the in vitro toxicity of three pristine ENPs (TiO2, Ag and SiO₂), three aged paints containing ENPs (TiO₂, Ag and SiO₂) and control paints without ENPs were compared. In a first experiment, cytotoxicity was assessed using a biculture consisting of human bronchial epithelial (16HBE14o-) cells and human monocytic cells (THP-1) to determine subtoxic concentrations. In a second experiment, a new coculture model of the lung-blood barrier consisting of 16HBE14o- cells, THP-1 and human lung microvascular endothelial cells (HLMVEC) was used to study pulmonary and extrapulmonary toxicity. The results show that the pristine TiO₂ and Ag ENPs have some cytotoxic effects at relative high dose, while pristine SiO₂ ENPs and all aged paints with ENPs and control paints do not. In the complex triculture model of the lung-blood barrier, no considerable changes were observed after exposure to subtoxic concentration of the different pristine ENPs and paint particles. In conclusion, we demonstrated that although pristine ENPs show some toxic effects, no significant toxicological effects were observed when they were embedded in a complex paint matrix., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)

Electronic noses/tongues (eN/eT) have emerged as promising alternatives for analysis of complex mixtures in the domain of food and beverage quality control. We have recently developed an electronic tongue by combining surface plasmon resonance imaging (SPRi) with an array of non-specific and cross-reactive receptors prepared by simply mixing two small molecules in varying and controlled proportions and allowing the mixtures to self-assemble on the SPRi prism surface. The obtained eT generated novel and unique 2D continuous evolution profiles (CEPs) and 3D continuous evolution landscapes (CELs) based on which the differentiation of complex mixtures such as red wine, beer and milk were successful. The preliminary experiments performed for monitoring the deterioration of UHT milk demonstrated its potential for quality control applications. Furthermore, the eT exhibited good repeatability and stability, capable of operating after a minimum storage period of 5 months., (Copyright © 2014 Elsevier B.V. All rights reserved.)

In Angiosperms, the biosynthesis of galactolipids involves enzymes localized in the inner envelope membrane (IEM) of chloroplasts, including a phosphatidic acid phosphatase (PAP), dephosphorylating phosphatidic acid (PA) into diacylglycerol (DAG), and MGD1, transferring a galactose onto DAG thus generating monogalactosyldiacylglycerol (MGDG). It has been shown that PA and DAG could be synthesized in the plastid via the so-called 'prokaryotic' pathway or imported from the endoplasmic reticulum via the 'eukaryotic' pathway. In vitro studies support the existence of (1) a negative regulation of the plastid PAP by DAG and (2) an activation of MGD1 by PA. We developed a mathematical model of the IEM galactolipid biosynthesis pathway to understand the properties of the system ruled by the presence of these two regulatory motifs. We demonstrated that the design of the system implies that PA should accumulate to levels that are not observed experimentally, regardless of its prokaryotic or eukaryotic origin. PA should therefore be used for other syntheses, such as that of phosphatidylglycerol. Whereas a massive influx of eukaryotic PA appears unlikely, an influx of eukaryotic DAG in the IEM is supported by simulations. The model also implies that DAG cannot transiently accumulate and that PA mainly acts as a signal switching the whole system on. Eventually, this analysis highlights the fact that the PAP enzyme could easily become dispensable and that the design of the system, with the two regulatory motifs, could precede the loss of the PAP gene or activity in this pathway, a phenomenon that occurred independently in most clades of Angiosperms., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Adducts formed at the nucleophilic N7 position of guanine are the most abundant lesions produced by alkylating agents such as ethylene oxide (EO) and propylene oxide (PO). In order to investigate the intrinsic mutagenic potential of N7-alkylguanine adducts, we prepared single-stranded DNA probes containing a single well-defined N7-alkylguanine adduct under conditions that minimize the presence of depurinated molecules. Following introduction of these probes into Escherichia coli cells, the effect of the N7-alkylguanine adducts on the efficiency and fidelity of replication was determined. To investigate the effect on replication we monitored the relative transformation efficiency of the lesion containing constructs with respect to the control construct. The methyl adduct was found not to be toxic, while the N7-(2-hydroxyethyl)guanine (N7-heG) and N7-(2-hydroxypropyl)guanine (N7-hpG) adducts reduce the transformation efficiency to ≈70% and 40%, respectively. Within the detection limits of our assay, replication across the N7-alkylguanine adducts in vivo is essentially error-free, as no mutant colony was observed among ≈300 individual sequenced colonies (i.e., mutation frequency<0.3%)., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Hydroxyl radical (OH) and one-electron oxidants that may be endogenously formed through oxidative metabolism, phagocytosis, inflammation and pathological conditions constitute the main sources of oxidatively generated damage to cellular DNA. It is worth mentioning that exposure of cells to exogenous physical agents (UV light, high intensity UV laser, ionizing radiation) and chemicals may also induce oxidatively generated damage to DNA. Emphasis is placed in this short review article on the mechanistic aspects of OH and one-electron oxidant-mediated formation of single and more complex damage (tandem lesions, intra- and interstrand cross-links, DNA-protein cross-links) in cellular DNA arising from one radical hit. This concerns DNA modifications that have been accurately measured using suitable analytical methods such as high performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry. Evidence is provided that OH and one-electron oxidants after generating neutral radicals and base radical cations respectively may partly induce common degradation pathways. In addition, selective oxidative reactions giving rise to specific degradation products of OH and one-electron oxidation reactions that can be used as representative biomarkers of these oxidants have been identified., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Taking the opportunity of the 20th anniversary of the word "proteomics", this young adult age is a good time to remember how proteomics came from enormous progress in protein separation and protein microanalysis techniques, and from the conjugation of these advances into a high performance and streamlined working setup. However, in the history of the almost three decades that encompass the first attempts to perform large scale analysis of proteins to the current high throughput proteomics that we can enjoy now, it is also interesting to underline and to recall how difficult the first decade was. Indeed when the word was cast, the battle was already won. This recollection is mostly devoted to the almost forgotten period where proteomics was being conceived and put to birth, as this collective scientific work will never appear when searched through the keyword "proteomics"., Biological Significance: The significance of this manuscript is to recall and review the two decades that separated the first attempts of performing large scale analysis of proteins from the solid technical corpus that existed when the word "proteomics" was coined twenty years ago. This recollection is made within the scientific historical context of this decade, which also saw the blossoming of DNA cloning and sequencing. This article is part of a Special Issue entitled: 20 years of Proteomics in memory of Viatliano Pallini. Guest Editors: Luca Bini , Juan J. Calvete, Natacha Turck, Denis Hochstrasser and Jean-Charles Sanchez., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Engineered TiO2 nanoparticles (TiO2-NPs) are present in a large variety of consumer products, and are produced in largest amount. The building industry is a major sector using TiO2-NPs, especially in paints. The fate of NPs after their release in the environment is still largely unknown, and their possible transfer in plants and subsequent impacts have not been studied in detail. The foliar transfer pathway is even less understood than the root pathway. In this study, lettuces were exposed to pristine TiO2-NPs and aged paint leachate containing TiO2-NPs and microparticles (TiO2-MPs). Internalization and in situ speciation of Ti were investigated by a combination of microscopic and spectroscopic techniques. Not only TiO2-NPs pristine and from aged paints, but also TiO2-MPs were internalized in lettuce leaves, and observed in all types of tissues. No change in speciation was noticed, but an organic coating of TiO2-NPs is likely. Phytotoxicity markers were tested for plants exposed to pristine TiO2-NPs. No acute phytotoxicity was observed; variations were only observed in glutathione and phytochelatin levels but remained low as compared to typical values. These results obtained on the foliar uptake mechanisms of nano- and microparticles are important in the perspective of risk assessment of atmospheric contaminations., (Copyright © 2014 Elsevier B.V. All rights reserved.)

5-Methylcytosine and methylated histones have been considered for a long time as stable epigenetic marks of chromatin involved in gene regulation. This concept has been recently revisited with the detection of large amounts of 5-hydroxymethylcytosine, now considered as the sixth DNA base, in mouse embryonic stem cells, Purkinje neurons and brain tissues. The dioxygenases that belong to the ten eleven translocation (TET) oxygenase family have been shown to initiate the formation of this methyl oxidation product of 5-methylcytosine that is also generated although far less efficiently by radical reactions involving hydroxyl radical and one-electron oxidants. It was found as additional striking data that iterative TET-mediated oxidation of 5-hydroxymethylcytosine gives rise to 5-formylcytosine and 5-carboxylcytosine. This survey focuses on chemical and biochemical aspects of the enzymatic oxidation reactions of 5-methylcytosine that are likely to be involved in active demethylation pathways through the implication of enzymatic deamination of 5-methylcytosine oxidation products and/or several base excision repair enzymes. The high biological relevance of the latter modified bases explains why major efforts have been devoted to the design of a broad range of assays aimed at measuring globally or at the single base resolution, 5-hydroxymethylcytosine and the two other oxidation products in the DNA of cells and tissues. Another critical issue that is addressed in this review article deals with the assessment of the possible role of 5-methylcytosine oxidation products, when present in elevated amounts in cellular DNA, in terms of mutagenesis and interference with key cellular enzymes including DNA and RNA polymerases., (Copyright © 2013 Elsevier B.V. All rights reserved.)

In this paper we review simulation and experimental studies of thermal capillary wave fluctuations as an ideal means for probing the underlying disjoining pressure and surface tensions, and more generally, fine details of the Interfacial Hamiltonian Model. We discuss recent simulation results that reveal a film-height-dependent surface tension not accounted for in the classical Interfacial Hamiltonian Model. We show how this observation may be explained bottom-up from sound principles of statistical thermodynamics and discuss some of its implications., (Copyright © 2013 Elsevier B.V. All rights reserved.)

The impact of engineered nanomaterials on plants, which act as a major point of entry of contaminants into trophic chains, is little documented. The foliar pathway is even less known than the soil-root pathway. However, significant inputs of nanoparticles (NPs) on plant foliage may be expected due to deposition of atmospheric particles or application of NP-containing pesticides. The uptake of Ag-NPs in the crop species Lactuca sativa after foliar exposure and their possible biotransformation and phytotoxic effects were studied. In addition to chemical analyses and ecotoxicological tests, micro X-ray fluorescence, micro X-ray absorption spectroscopy, time of flight secondary ion mass spectrometry and electron microscopy were used to localize and determine the speciation of Ag at sub-micrometer resolution. Although no sign of phytotoxicity was observed, Ag was effectively trapped on lettuce leaves and a thorough washing did not decrease Ag content significantly. We provide first evidence for the entrapment of Ag-NPs by the cuticle and penetration in the leaf tissue through stomata, for the diffusion of Ag in leaf tissues, and oxidation of Ag-NPs and complexation of Ag(+) by thiol-containing molecules. Such type of information is crucial for better assessing the risk associated to Ag-NP containing products., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Radiosensitization efficacy of gold nanoparticles (AuNPs) with low energy radiations (88 keV) was evaluated in vitro and in vivo on rats bearing glioma. In vitro, a significant dose-enhancement factor was measured by clonogenic assays after irradiation with synchrotron radiation of F98 glioma cells in presence of AuNPs (1.9 and 15 nm in diameter). In vivo, 1.9 nm nanoparticles were found to be toxic following intracerebral delivery in rats bearing glioma, whether no toxicity was observed using 15 nm nanoparticles at the same concentration (50 mg/mL). The therapeutic efficacy of gold photoactivation was determined by irradiating the animals after intracerebral infusion of AuNPs. Survival of rats that had received the combination of treatments (AuNPs: 50 mg/mL, 15 Gy) was significantly increased in comparison with the survival of rats that had received irradiation alone. In conclusion, this experimental approach is promising and further studies are foreseen for improving its therapeutic efficacy., From the Clinical Editor: These investigators report that gold nanoparticles of the correct size can be used to enhance the effects of irradiation in the context of a glioma model. Since many of the glioma varieties are currently incurable, this or similar approaches may find their way to clinical trials in the near future., (Copyright © 2013 Elsevier Inc. All rights reserved.)

One of the challenges of the proteomic analysis by 2D-gel is to visualize the low abundance proteins, particularly those localized in the organelles. An additional problem with nuclear proteins lies in their strong interaction with nuclear acids. Several experimental procedures have been tested to increase, in the nuclear extract, the ratio of nuclear proteins compared to contaminant proteins, and also to obtain reproducible conditions compatible with 2D-gel electrophoresis. The NaCl procedure has been chosen. To test the interest of this procedure, the nuclear protein expression profiles of macrophages and dendritic cells have been compared with a proteomic approach by 2D-gel electrophoresis. Delta2D software and mass spectrometry analyses have allowed pointing out some proteins of interest. We have chosen some of them, involved in transcriptional regulation and/or chromatin structure for further validations. The immunoblotting experiments have shown that most of the observed changes are due to post-translational modifications, thereby exemplifying the interest of the 2D gel approach. Finally, this approach allowed us to reach not only high abundance nuclear proteins but also lower abundance proteins, such as the HP1 proteins and reinforces the interest of using 2DE-gel in proteomics because of its ability to visualize intact proteins with their modifications., (Copyright © 2012 Elsevier B.V. All rights reserved.)

A nuclear magnetic resonance imaging technique is used to measure velocity distributions in turbulent pipe flows up to Re=24580. While turbulent intensity is usually determined from signal attenuation, we deduce turbulent intensity from velocity distribution with no need to suppose a gaussian distribution for velocity fluctuations. Skewness and flatness measurements are also presented in this paper. Comparison with DNS show good agreement and we show that NMR data is sufficently accurate to provide turbulent viscosity profile. The low field system used in this study allow the suppression of susceptibility artifacts and thus open its use for studying two-phase flows. We postulate that the method used here could be applied to two-phase flows and would thus provide valuable information on turbulent viscosity models., (Copyright © 2012 Elsevier Inc. All rights reserved.)

The accumulation of amyloid fibers due to protein misfolding is associated with numerous human diseases. For example, the formation of amyloid deposits in neurodegenerative pathologies is correlated with abnormal apoptosis. We report here the in vitro formation of various types of aggregates by Bcl-xL, a protein of the Bcl-2 family involved in the regulation of apoptosis. Bcl-xL forms aggregates in three states, micelles, native-like fibrils, and amyloid fibers, and their biophysical characterization has been performed in detail. Bcl-xL remains in its native state within micelles and native-like fibrils, and our results suggest that native-like fibrils are formed by the association of micelles. Formation of amyloid structures, that is, nonnative intermolecular β-sheets, is favored by the proximity of proteins within fibrils at the expense of the Bcl-xL native structure. Finally, we provide evidence of a direct relationship between the amyloid character of the fibers and the tertiary-structure stability of the native Bcl-xL. The potential causality between the accumulation of Bcl-xL into amyloid deposits and abnormal apoptosis during neurodegenerative diseases is discussed., (Copyright © 2011. Published by Elsevier Ltd.)

Two-dimensional electrophoresis of proteins has preceded, and accompanied, the birth of proteomics. Although it is no longer the only experimental scheme used in modern proteomics, it still has distinct features and advantages. The purpose of this tutorial paper is to guide the reader through the history of the field, then through the main steps of the process, from sample preparation to in-gel detection of proteins, commenting the constraints and caveats of the technique. Then the limitations and positive features of two-dimensional electrophoresis are discussed (e.g. its unique ability to separate complete proteins and its easy interfacing with immunoblotting techniques), so that the optimal type of applications of this technique in current and future proteomics can be perceived. This is illustrated by a detailed example taken from the literature and commented in detail. This Tutorial is part of the International Proteomics Tutorial Programme (IPTP 2)., (Copyright © 2011 Elsevier B.V. All rights reserved.)

This survey focuses on the critical evaluation of the main methods that are currently available for monitoring single and complex oxidatively generated damage to cellular DNA. Among chromatographic methods, HPLC-ESI-MS/MS and to a lesser extent HPLC-ECD which is restricted to a few electroactive nucleobases and nucleosides are appropriate for measuring the formation of single and clustered DNA lesions. Such methods that require optimized protocols for DNA extraction and digestion are sensitive enough for measuring base lesions formed under conditions of severe oxidative stress including exposure to ionizing radiation, UVA light and high intensity UVC laser pulses. In contrast application of GC-MS and HPLC-MS methods that are subject to major drawbacks have been shown to lead to overestimated values of DNA damage. Enzymatic methods that are based on the use of DNA repair glycosylases in order to convert oxidized bases into strand breaks are suitable, even if they are far less specific than HPLC methods, to deal with low levels of single modifications. Several other methods including immunoassays and (32)P-postlabeling methods that are still used suffer from drawbacks and therefore are not recommended. Another difficult topic is the measurement of oxidatively generated clustered DNA lesions that is currently achieved using enzymatic approaches and that would necessitate further investigations., (2011 Elsevier B.V. All rights reserved.)

On the basis of labeling experiments, Grove et al. (2011) have shown how an electrophilic carbon (from an RNA adenosine) can be methylated by S-adenosylmethionine-dependent methyltransferases though an original radical mechanism., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Platinating agents are commonly prescribed anticancer drugs damaging DNA. Induced lesions are recognized by a wide range of proteins. These are involved in cellular mechanisms such as DNA repair, mediation of cytotoxicity or chromatin remodeling. They therefore constitute crucial actors to understand pharmacology of these drugs. To expand our knowledge about this subproteome, we developed a ligand fishing trap coupled to high throughput proteomic tools. This trap is made of damaged plasmids attached to magnetic beads, and was exposed to cell nuclear extracts. Retained proteins were identified by nanoHPLC coupled to tandem mass spectrometry. This approach allowed us to establish a list of 38 proteins interacting with DNA adducts generated by cisplatin, oxaliplatin and satraplatin. Some of them were already known interactome members like high mobility group protein 1 (HMGB1) or the human upstream binding factor (hUBF), but we also succeeded in identifying unexpected proteins such as TOX HMG box family member 4 (TOX4), phosphatase 1 nuclear targeting subunit (PNUTS), and WD repeat-containing protein 82 (WDR82), members of a recently discovered complex. Interaction between TOX4 and platinated DNA was subsequently validated by surface plasmon resonance imaging (SPRi). These interactions highlight new cellular responses to DNA damage induced by chemotherapeutic agents., (Copyright © 2010 Elsevier Inc. All rights reserved.)

Dark field electron holography is a new TEM-based technique for measuring strain with nanometer scale resolution. Here we present the procedure to align a transmission electron microscope and obtain dark field holograms as well as the theoretical background necessary to reconstruct strain maps from holograms. A series of experimental parameters such as biprism voltage, sample thickness, exposure time, tilt angle and choice of diffracted beam are then investigated on a silicon-germanium layer epitaxially embedded in a silicon matrix in order to obtain optimal dark field holograms over a large field of view with good spatial resolution and strain sensitivity., (Copyright © 2010 Elsevier B.V. All rights reserved.)

Exposure to polycyclic aromatic hydrocarbons (PAHs) always involves complex mixtures that may induce synergistic or antagonistic effects on the genotoxic properties and make risk assessment more difficult. In this study, we evaluated how particulate PAHs modulated the formation of DNA damage induced by carcinogenic benzo[a]pyrene (B[a]P). Single strand breaks and alkali labile sites, as well as BPDE-N²-dGuo DNA adducts were measured in the competent HepG2 cells by Comet assay and HPLC-tandem mass spectrometry, respectively. B[a]P, alone or in binary mixture with other PAHs (1 μM each), led to low amounts of strand breaks. In contrast, formation of BPDE-N²-dGuo adducts was significant and found to be enhanced in HepG2 co-treated for 14 h by B[a]P in the presence of either benzo[b]fluoranthene (B[b]F), dibenz[a,h]anthracene (DB[a,h]A) or indeno[1,2,3-cd]pyrene (IP). Opposite results were obtained with benzo[k]fluoranthene (B[k]F). The same observations were made when cells were pre-incubated with PAH before incubation with B[a]P. These results show that the interactions between PAHs are not direct competition reactions. Emphasis was then placed on the modulation of B[a]P-induced DNA damage by B[b]F and B[k]F. No difference in the time-course formation of DNA damage was observed. However, dose-response relationship differed between these two PAHs with a concentration-dependent inhibition of BPDE-N²-dGuo DNA by B[k]F whereas a constant level of potentiation for B[b]F was observed for concentrations higher than 1 μM. Altogether, these results show that the genotoxicity of B[a]P in binary mixtures with other carcinogenic PAH may be modulated. In such cases, a potentiation of BPDE-N²-dGuo adduct formation is most often observed with exception of B[k]F. Several biological mechanisms may account for these observations, including binding of PAHs to the Ah receptor (AhR), their affinity toward CYP450 and competition for metabolism. These different interactions have to be considered when addressing the intricate issue of the toxicity of mixtures., (Copyright © 2010 Elsevier Ireland Ltd. All rights reserved.)

Aminodeoxychorismate (ADC) synthase in plants is a bifunctional enzyme containing glutamine amidotransferase (GAT) and ADC synthase (ADCS) domains. The GAT domain releases NH(3) from glutamine and the ADCS domain uses NH(3) to aminate chorismate. This enzyme is involved in folate (vitamin B9) biosynthesis. We produced a stable recombinant GAT-ADCS from Arabidopsis. Its kinetic properties were characterized, and activities and coupling of the two domains assessed. Both domains could operate independently, but not at their optimal capacities. When coupled, the activity of one domain modified the catalytic properties of the other. The GAT activity increased in the presence of chorismate, an activation process that probably involved conformational changes. The ADCS catalytic efficiency was 10(4) fold higher with glutamine than with NH(4)Cl, indicating that NH(3) released from glutamine and used for ADC synthesis did not equilibrate with the external medium. We observed that the GAT activity was always higher than that of ADCS, the excess of NH(3) being released in the external medium. In addition, we observed that ADC accumulation retro-inhibited ADCS activity. Altogether, these results indicate that channeling of NH(3) between the two domains and/or amination of chorismate are the limiting step of the whole process, and that ADC cannot accumulate., (Copyright © 2010 Elsevier Inc. All rights reserved.)

Anti-angiogenic therapies of solid cancers aim at specifically destroying the tumor vasculature in order to "asphyxiate" the tumors. Since few years, they represent a novel therapeutic tool, which allowed to significantly improve the survival of patients suffering from colon, breast, kidney and lung cancers. However, these therapies are limited in their efficacy by the appearance of tumor resistance phenomena. In this review, I describe the molecular and cellular mechanisms of tumor angiogenesis with a special focus on the important roles played by hypoxia, the endothelial growth factor VEGF and the endothelial tip-cells located at the extremity of sprouting neo-vessels. I present the factors that respectively control the activation phase and the maturation phase of angiogenesis, as well as their mechanisms of action. In a second part, the efficacy and the limits of anti-angiogenic therapies presently available on the market are described, and the recent elucidation of some molecular mechanisms of tumor resistance to anti-angiogenic therapies is reviewed.

Two-dimensional gel electrophoresis has been instrumental in the birth and developments of proteomics, although it is no longer the exclusive separation tool used in the field of proteomics. In this review, a historical perspective is made, starting from the days where two-dimensional gels were used and the word proteomics did not even exist. The events that have led to the birth of proteomics are also recalled, ending with a description of the now well-known limitations of two-dimensional gels in proteomics. However, the often-underestimated advantages of two-dimensional gels are also underlined, leading to a description of how and when to use two-dimensional gels for the best in a proteomics approach. Taking support of these advantages (robustness, resolution, and ability to separate entire, intact proteins), possible future applications of this technique in proteomics are also mentioned., (Copyright © 2010 Elsevier B.V. All rights reserved.)

Electrophoretic separations of proteins are widely used in proteomic analyses, and rely heavily on SDS electrophoresis. This mode of separation is almost exclusively used when a single dimension separation is performed, and generally represents the second dimension of two-dimensional separations. Electrophoretic separations for proteomics use robust, well-established protocols. However, many variations in almost all possible parameters have been described in the literature over the years, and they may bring a decisive advantage when the limits of the classical protocols are reached. The purpose of this article is to review the most important of these variations, so that the readers can be aware of how they can improve or tune protein separations according to their needs. The chemical variations reviewed in this paper encompass gel structure, buffer systems and detergents for SDS electrophoresis, two-dimensional electrophoresis based on isoelectric focusing and two-dimensional electrophoresis based on cationic zone electrophoresis., (Copyright 2010 Elsevier B.V. All rights reserved.)

Yeast ubiquinone or coenzyme Q(6) (Q(6)) is a redox active lipid that plays a crucial role in the mitochondrial electron transport chain. At least nine proteins (Coq1p-9p) participate in Q(6) biosynthesis from 4-hydroxybenzoate (4-HB). We now show that the mitochondrial ferredoxin Yah1p and the ferredoxin reductase Arh1p are required for Q(6) biosynthesis, probably for the first hydroxylation of the pathway. Conditional Gal-YAH1 and Gal-ARH1 mutants accumulate 3-hexaprenyl-4-hydroxyphenol and 3-hexaprenyl-4-aminophenol. Para-aminobenzoic acid (pABA) is shown to be the precursor of 3-hexaprenyl-4-aminophenol and to compete with 4-HB for the prenylation reaction catalyzed by Coq2p. Yeast cells convert U-((13)C)-pABA into (13)C ring-labeled Q(6), a result that identifies pABA as a new precursor of Q(6) and implies an additional NH(2)-to-OH conversion in Q(6) biosynthesis. Our study identifies pABA, Yah1p, and Arh1p as three actors in Q(6) biosynthesis., (2010 Elsevier Ltd. All rights reserved.)

Polycyclic aromatic hydrocarbons (PAH) produced upon incomplete combustion of organic matter are suspected to be carcinogenic to humans. In the present work, we especially studied the genotoxicity of benzo[a]pyrene (B[a]P), pure or in mixtures, with emphasis placed on the contribution of oxidative stress and alkylation. A comparison was made between the extent of DNA strand breaks as determined by the Comet assay and the number of DNA adducts to the diol epoxide metabolite of B[a]P measured by HPLC-mass spectrometry. HepG2 cultured human hepatocytes were treated with either pure B[a]P or particulate matter extracted from air samples collected in an urban peri-industrial site or in a metallurgic plant. Treatment with pure B[a]P did not induce increase in Comet measurements below a concentration of 1 microM whereas adducts were observed for concentrations as low as 0.025 microM. Very different results were obtained with environmental samples. Increase in the Comet score was observed with both urban and industrial mixtures containing 0.16 microM of B[a]P, especially for samples of urban origin. Comparison with the effect of the reconstituted PAH fraction of the mixtures allowed us to conclude that the induction of strand breaks results from the action of other components of the samples. In addition, a 30% potentialization and a 90% inhibition in the level of DNA adducts with respect to exposure to 0.16 microM pure B[a]P were observed for cells exposed to industrial and urban mixtures, respectively. These results contrast with the 6-fold enhancement in the yield of BPDE adducts in cells exposed to the reconstituted PAH fraction with respect to pure BaP. Altogether, our data emphasize that (i) a combination of analytical approaches is required to assess the genotoxicity of complex mixtures and (ii) risk assessment based on additivity consideration such as toxic equivalent factors may be misleading.

Cyclobutane pyrimidine dimers (CPDs) and pyrimidine (6-4) pyrimidone photoproducts (6-4PPs) are the two main classes of mutagenic DNA damages induced by UVB radiation. Numerous studies have been devoted so far to their formation and repair in human cells and skin. However, the biochemical methods used often lack the specificity that would allow the individual study of each of the four CPDs and 6-4PPs produced at TT, TC, CT and CC dinucleotides. In the present work, we applied an HPLC-mass spectrometry assay to study the formation and repair of CPDs and 6-4PPs photoproducts in primary cultures of human keratinocytes and fibroblasts as well as in whole human skin. We first observed that the yield of dimeric lesions was slightly higher in fibroblasts than in keratinocytes. In contrast, the rate of global repair was higher in the last cell type. Moreover, removal of DNA photoproducts in skin biopsies was found to be slower than in both cultured skin cells. In agreement with previous works, the repair of 6-4PPs was found to be more efficient than that of CPDs in the three types of samples, with no observed difference between the removal of the TT and TC derivatives. In contrast, a significant influence of the nature of the two modified pyrimidines was observed on the repair rate of CPDs. The decreasing order of removal efficiency was the following: C<>T>C<>C>T<>C>T<>T. These data, together with the known intrinsic mutational properties of the lesions, would support the reported UV mutation spectra. A noticeable exception concerns CC dinucleotides that are mutational hotspots with an UV-specific CC to TT tandem mutation, although related bipyrimidine photoproducts are produced in low yields and efficiently repaired.

Numerous data show a functional link between lymphangiogenesis, lymph node invasion by tumoral cells and metastasis. During the last decade, the identification of lymphatic endothelial cell-specific markers has allowed the investigation of lymphangiogenesis regulatory mechanisms and the analysis of its involvement in tumoral progression. Among regulatory systems, the growth factors VEGF-C and D, that bind and activate their common receptor VEGFR3, appear to play an important role in this process. Therapeutic strategies targeting this pathway or, in a general manner, aiming at inhibiting tumoral lymphangiogenesis are now considered to block the development of tumoral metastasis. Further fundamental and clinical studies are clearly needed to establish the pronostic value of lymphangiogenesis and to validate anti-lymphangiogenic therapies in the treatment of metastatic cancers.

Copper is an essential trace element for successful pregnancy. However, the mechanisms by which copper is transported from maternal circulation to the fetus have not been clearly elucidated. Two proteins, cellular prion (PrP(C)) and COMMD1, are known to be responsible for prion diseases and canine copper toxicosis, respectively, and are thought to play a role in copper homeostasis. However, their placental expression and localization throughout human gestation are still unknown. In this study, we used quantitative RT-PCR, western blotting and immunohistochemistry to investigate in detail the expression and localization of PrP(C) and COMMD1 proteins in human placenta throughout pregnancy. Our results show that both proteins are expressed in human placenta. PrP(C) showed the highest mRNA and protein expression levels during the first trimester of pregnancy. PrP(C) and COMMD1 proteins are similarly localized within the placental villi. Both proteins are present in the syncytiotrophoblast, the cytotrophoblast, vascular endothelial cells and Hofbauer cells. These data offer some insights into possible roles for PrP(C) and COMMD1 within the placenta.

The dynamic behavior of water within two types of ionomer membranes, Nafion and sulfonated polyimides, has been investigated by field-cycling nuclear magnetic relaxation. This technique, applied to materials prepared at different hydration levels, allows to probe the proton motion on a time scale of the microsecond. The NMR longitudinal relaxation rate R(1) measured over three decades of Larmor angular frequencies omega is particularly sensitive to the host-water interactions and thus well suited to study fluid dynamics in restricted geometries. In the polyimide membranes, we have observed a strong dispersion of R(1)(omega) following closely a 1/sqrt[omega] law in a low-frequency range (correlation times from 0.1 to 10 micros). This is indicative of a strong interaction of water with "interfacial" hydrophilic groups of the polymeric matrix (wetting situation). On the contrary, in the Nafion, we observed weak variations of R(1)(omega) at low frequency. This is typical of a nonwetting behavior. At early hydration stages, the proton-proton inter-dipolar contribution to R(1)(omega) evolves logarithmically, suggesting a confined bidimensional diffusion of protons in the microsecond time range. Such an evolution is lost at higher swelling where a plateau related to 3D diffusion is observed.

Propagation of the scrapie isoform of the prion protein (PrP(Sc)) depends on the expression of endogenous cellular prion (PrP(C)). During oral infection, PrP(Sc) propagates, by conversion of the PrP(C) to PrP(Sc), from the gastrointestinal tract to the nervous system. Intestinal epithelium could serve as the primary site for PrP(C) conversion. To investigate PrP(C) sorting in epithelia cells, we have generated both a green fluorescent protein (EGFP) or hemagglutinin (HA) tagged human PrP(C) (hPrP(C)). Combined molecular, biochemical, and single living polarized cell imaging characterizations suggest that hPrP(C) is selectively targeted to the apical side of Madin-Darby canine kidney (MDCKII) and of intestinal epithelia (Caco2) cells.

Alix is a phylogenetically conserved protein that participates in mammals in programmed cell death in association with ALG-2, a penta-EF-hand calciprotein. It contains an N-terminal Bro1 domain, a coiled-coil region and a C-terminal proline-rich domain containing several SH3- and WW-binding sites that contribute to its scaffolding properties. Recent data showed that by virtue of its Bro1 domain, Alix is functionally associated to the ESCRT complexes involved in the biogenesis of the multivesicular body and sorting of transmembrane proteins within this specific endosomal compartment. In Dictyostelium, an alx null strain shows a markedly perturbed starvation-induced morphogenetic program while ALG-2 disruptants remain unaffected. This review summarizes Dictyostelium data on Alix and ALG-2 homologues and evaluates whether known functions of Alix in other organisms can account for the developmental arrest of the alx null mutant and how Dictyostelium studies can substantiate the current understanding of the function(s) of this versatile and conserved signaling molecule.

Using a radial flow chamber, we study Saccharomyces cerevisiae kinetics of detachment from stainless steel substrates. Samples of similar surface chemistry, but with different surface topologies are compared: mirror polished and electro-chemically etched. Different grain sizes (20, 40 and 100 microm) and different etching depths (100-650 nm) are tested. Cells are removed from the substrate according to a first-order kinetics defining two macroscopic parameters that depend on the applied stress: the detachment efficiency and the detachment rate constant. Whatever the surface topology, detachment occurs above a threshold and its rate is strongly stimulated by the applied stress. The detachment efficiency is characterized by the shear stress at which half of the cells detach and is independent of surface topology. In contrast, detachment is faster from etched than mirror polished surfaces. Finally, we also show the preferential adhesion of yeast cells to grains of < 001 > crystallographic orientation with respect to the surface.

Among the different trophoblast subtypes of the mouse placenta, the glycogen cells (GC) are one of the trophoblast subtypes that invade the decidua. We previously established that GC specifically expressed protocadherin 12 (PCDH12). In this paper, we investigated the origin of the PCDH12-positive cells and we characterized their fate in the maternal tissues. Our data indicate that they directly originate from the central part of the ectoplacental cone at embryonic day (E) 7.5. PCDH12-positive cells start to accumulate glycogen from E10.5 and the first migrating cells could be observed from this age. Unlike other placental and decidual cells, GC do not express P-cadherin, which may explain their migration properties in this organ. In the decidua, GC settle in the vicinity of the maternal vascular sinuses but do not incorporate in the endothelium. By the end of gestation (E17.5), most GC islets of the decidua enter into a lytic phase and form large lacunae. These lacunae, filled with glycogen, may provide a substantial source of energy at the end of gestation or during delivery. Our data suggest that spongiotrophoblasts and GC are two independent lineages and we bring insights into GC migration and fate.

Background: It is debated whether brain death (BD) causes transient functional ischemia. In this investigation we used monophasic action potential (AP) recording during BD as a sensitive means to assess: (i) whether ischemia was present; and (ii) the effect of BD on a subsequent ischemia-reperfusion challenge., Methods: In Period 1, BD was induced (BD group, 6 pigs) or not induced (sham maneuver, control [C] group, 6 pigs), and effects were followed for 3 hours. In Period 2, left anterior descending (LAD) coronary artery ligation ischemia was applied for 20 minutes to all hearts, followed by 60-minute reperfusion., Results: In Period 1, plasma norepinephrine was 3.1-, 6.3- and 5-fold greater in BD than in C at 1, 120 and 180 minutes, respectively, and systolic blood pressure was 26% greater at 1 minute and 35% at 120 minutes. The arteriovenous difference in lactate was similar or lower in BD than in C. In both groups, at all time-points, the action potential recording had a rectangular plateau shape and action potential duration (APD50) had a linear relationship to the cardiac inter-beat (RR) interval (R2 = 0.89 and 0.73, slope = 0.42 +/- 0.02 and 0.46 +/- 0.06 in BD and C, respectively). In Period 2, ischemia caused a similar (50%) APD shortening in BD and C. Restoration of the APD upon reperfusion was complete in both groups., Conclusions: Our findings suggest that BD does not cause direct cardiac ischemia and does not change the response of the heart to subsequent ischemia-reperfusion challenge.

The Proteus mirabilis catalase is one of the most efficient heme-containing catalase and forms a relatively stable compound II. Samples of compound II were prepared from PMC enriched in (57)Fe. For the first time, two different forms of compound II, namely low pH compound II (LpH II) (43%) and high pH compound II (HpH II) (25%), have been characterized by Mössbauer spectroscopy at pH 8.3. The ratio LpH II/HpH II increases irreversibly with decreasing pH. The large quadrupole splitting value of LpH II (DeltaE(Q)=2.29 (2) mm/s, with delta(/Fe)=0.03 (2) mm/s), compared to that of HpH II (DeltaE(Q)=1.47 (2) mm/s, with delta(/Fe)=0.07 (2) mm/s), reflects the protonation of the ferryl group. Quadrupole splitting values of 1.46 and 2.15mm/s have been computed by DFT for optimized models of the ferryl compound II (model 1) and the protonated ferryl compound II (model 2), respectively, starting from the Fe(IV)O model initially published by Rovira and Fita [C. Rovira, I. Fita, J. Phys. Chem. B 107 (2003) 5300-5305]. Therefore, we attribute the LpH II compound to a protonated ferryl Fe(IV)-OH complex, whereas the HpH II compound corresponds to the classical ferryl Fe(IV)O complex.

The Geometrical phase analysis, which is a very efficient method to measure deformation from High resolution transmission electron microscopy images, is studied from a theoretical point of view. We point out that the basic property of this method is its ability to measure local reciprocal lattice parameters with a high level of accuracy. We attempt to provide some insights into (a) different formula used in the geometrical phase analysis such as the well-known relation between phase and displacement: Pg(r)=-2pi g.u(r), (b) the two different definitions of strain, each of which corresponding to a different lattice reference and (c) the meaning of a continuous displacement in a dot-like high resolution image. The case of one-dimensional analysis is also presented. Finally, we show that the method is able to give the position of the dot that is nearest to a given pixel in the image.