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SummaryCulicoides boyi Nielsen, Kristensen & Pape 2015 (Diptera: Ceratopogonidae), described in Denmark in 2015, is reported in France. This species is closely related to Culicoides pulicaris (Linnaeus 1758) and can only be distinguished from this species by a subtle variation in spot wing pattern and by calculation of maxillary palpal ratio.

Culicoides (Diptera: Ceratopogonidae) serve as vectors of several mammalian and avian diseases, including bluetongue, Schmallenberg, African horse sickness, avian malaria and Oropouche. Host preference investigations are necessary to assess the transmission routes of vector-borne diseases and to inform mitigation strategies. A recent study examining the main sensory structures (palps and antennae) of Culicoides species suggests that they be classified as ornithophilic or mammalophilic according to their feeding habits. We analyzed Culicoides host preferences according to the literature and carried out a multiple correspondence analysis linking these preferences with morphological data. Seven out of 12 variables were found to be reliable predictors of host preference in Culicoides species: Antenna Flagellomer-Sensilla Coeloconica-Number: (7–10) and (11–13); Antenna Flagellomer-Sensilla Coeloconica IV–X: presence; Palpus-size: wide and/or narrow opening and shallow pit; Palpus-Shape: strongly swollen; Antenna-Short sensilla trichodea-distal part segment IV to X-Number: 2 seta each. Our results demonstrate that the presence of sensilla coeloconica and the maxillary palpus can be used to separate ornithophilic and mammalophilic or ornithophilic/mammalophilic species.

We attempted to characterize the cellular autofluorescence phenomenon of living HL-60 cells and to appraise its modifications under oxidative stress conditions induced by 1α,25(OH)2D3 (VD3) and its analog EB1089. Autofluorescence emission spectra of human promyelocytic HL-60 leukemic cells were monitored using laser scanning confocal microspectrofluorometry under UV excitation. Evaluation of reactive oxygen species (ROS) release was performed using the 2′,7′-dichlorodihydrofluorescein diacetate (H2-DCFDA) staining and fluorescence emission measurement. VD3 (1, 10, 100 nM) or EB1089 (0.1, 1 and 10 nM) induces a decrease in autofluorescence emission intensity that can be attributed to the oxidation of the coenzyme nicotinamide adenine dinucleotide (phosphate) NAD(P)H into NAD(P)+. A dose-dependent increase (p3- and EB1089-treated cells. As compared with VD3- or EB1089-treated cells, doxorubicin-VD3 or doxorubicin-EB1089 treatments strongly decrease the autofluorescence intensity and induce a higher release of ROS (p3 or EB1089 induce a more limited autofluorescence decrease and a weaker ROS generation, as compared with VD3 and EB1089 treated cells. In conclusion, the free radicals release, generated by VD3 and EB1089, was associated with the decrease in autofluorescence emission and can be modulated by doxorubicin and antioxidants.

Image cytometry was applied to study the intracellular localization of autofluorescence and the influence of an oxidative stress on this emission. K562 erythroleukemia cancer cells were analyzed with a microspectrofluorometer, coupled with a Argon laser (Ar+) (363 nm). From each cell, 15 x 15 emission spectra were recorded in the 400-600 nm spectral range to generate a spectral image of autofluorescence. The intracellular locations of the autofluorescence emission and of the specific mitochondrial probe rhodamine 123 (R123) were matched. Under a 363 nm excitation, all spectra from K562 cells show equivalent profiles with a 455 nm maximum emission, near of reduced nicotinamide adenine dinucleotide-(Phosphate) solution (NAD(P)H) (465 nm maximum emission). The spatial distribution of autofluorescence is homogeneous and different from the one of R123. Hydrogen peroxide (H2O2) (200 microM) and menadione (Men) (5 microM) induce a weak spectral change and a decrease in autofluorescence intensity, down to 40% of the initial emission. Doxorubicin (Dox) induces a dose-dependent decrease in autofluorescence emission and a release of intracellular free radicals. When cells were pre-treated 1 h with 1 mM glutathione (GSH), Dox induces a lower free radicals release, no significant variation of autofluorescence intensity and a lower growth inhibitory effect. Images cytometry of autofluorescence suggest that the intracellular NAD(P)H would not be restricted to mitochondrial compartments. The release of free radicals was associated with a decrease in autofluorescence intensity, mainly attributed to NAD(P)H oxidation both inside and outside mitochondria.

Fluorescence correlation spectroscopy (FCS) has been used to analyze the plasma membrane fluidity and heterogene- ity of multidrug resistant cells. At the single cell level, the effects of different membrane agents present in the extra-cellular medium have been explored. Firstly, we reveal a modification of plasma membrane heterogeneities according to the addition of a fluidity modulator, benzyl alcohol. On the other hand, revertants such as verapamil and cyclosporin-A appear to act more specifically on the slow diffusion sites, such as lipids microdomains.

National audience; La surface d'un substrat de cuivre a été fonctionnalisée grâce à une technique d'auto-assemblage d'une monocouche d'acide phosphonique. Après caractérisation de ce film, un excellent comportement tribologique est noté lorsqu'aucun nettoyage ultrasonore n'est effectué, pour des conditions de sollicitation de pression de 133 MPa et de vitesse de glissement de 1mm/s sur un tribomètre alternatif linéaire utilisant un frotteur en Si3N4 (μ = 0.12, durée de vie multipliée par 100). Des espèces physisorbées, sous formes d'amas de plusieurs μm3, présentes sur la monocouche semblent responsables de ce bon comportement. De premiers essais sur un tribomètre d'étirage-plan ont montré un réel potentiel de ce type de fonctionnalisation en emboutissage.

International audience; Diffusion-time distribution analysis (DDA) has been used to explore the plasma membrane fluidity of multidrug-resistant cancer cells (LR73 carcinoma cells) and also to characterize the influence of various membrane agents present in the extracellular medium. DDA is a recent single-molecule technique, based on fluorescence correlation spectroscopy (FCS), well suited to retrieve local organization of cell membrane. The method was conducted on a large number of living cells, which enabled us to get a detailed overview of plasma membrane microviscosity, and plasma membrane micro-organization, between the cells of the same line. Thus, we clearly reveal the higher heterogeneity of plasma membrane in multidrug-resistant cancer cells in comparison with the nonresistant ones (denoted sensitive cells). We also display distinct modifications related to a membrane fluidity modulator, benzyl alcohol, and two revertants of multidrug resistance, verapamil and cyclosporin-A. A relation between the distribution of the diffusion-time values and the modification of membrane lateral heterogeneities is proposed

Numerical modelling of the deformation of a polymer using the finite elements method in axisymetrical mode was performed using the LsDyna® software to describe the filling of micro-cavities during the forming process of the material using the hot embossing. These simulations firstly allow verifying whether the chosen forming process conditions promote or not an optimized filling of the superficial cavities in order to achieve precise replicas which best reproduce the superficial topography of the mould. The simulations were carried out to evaluate the filling of the cavities taking into account the mechanical behaviour of the selected polymer into the model. Moreover, these models were developed to verify the effect of the distribution of the mould cavities on their filling. The influence of the mobility of non deformable rigid plates on the filling of the cavities represents an auxiliary variable. In the approach presented, the compression plates are assumed to be parallel and non deformable, whereas the polymer disk follows a rubbery behaviour around a temperature equal to 140°C. Globally the modelling results are satisfactory for they are rather close to the experimental observations conducted. In summary, the effect of the normal stress as also the distribution of micro-cavities at the mould surface seem to prevail in the case of the forming process by hot embossing.

Fluorescence correlation spectroscopy (FCS) has been used to analyze the plasma membrane fluidity and heterogeneity of multidrug resistant cells. At the single cell level, the effects of different membrane agents present in the extra-cellular medium have been explored. Firstly, we reveal a modification of plasma membrane heterogeneities according to the addition of a fluidity modulator, benzyl alcohol. On the other hand, revertants such as verapamil and cyclosporin-A appear to act more specifically on the slow diffusion sites, such as lipids microdomains.

1alpha,25-dihydroxyvitamin D3 (VD3) and the EB1089 analog are well known for their roles in the modulation of proliferation and the differentiation of several malignant cells. In addition, VD3 or EB1089 displayed a high disposal of oxidant features and the ability to cause release of reactive oxygen species (ROS). We attempted to enhance HL60 cell differentiation and to limit ROS generation, by the association of deltanoids with doxorubicin and the antioxidants catalase (CAT), superoxide dismutase (SOD) and N-acetyl cystein (NAC). Differentiation of HL60 cells into monocytic lineage was studied by expression of mRNA, protein CD14 and functional differentiation by the nitroblue tetrazolium assay. The 2',7'-dichlorodihydrofluorescein diacetate (H2-DCFDA) dye allowed to evaluate in situ ROS generation. When associated with 0.1 nM EB1089, 15 nM doxorubicin induced an increase of differentiated cell percentage from 29% to 87% and did not affect VD3-treated cells. The association with doxorubicin also induced a significant increase of ROS release (p

In that paper, one uses the concepts of contact mechanics to describe the quality of reproduction of cylindrical cavity shape by hot embossing. It results in a negative replica of the initial shape. This model takes into account the deformation of the polymer material imposed by the forming process. These results point out the influence of experimental parameters (temperature, pressure) as well as those of the elastic modulus (Young’s modulus and Poisson’s ratio). The analytical data are compared with the metrology results obtained using a scanning mechanical microscope in two or three‐dimensional mode. The set of data enables a predictive approach of the engraving quality depending on the polymer mechanical properties. The final goal is to adapt this model to the hot embossing process.

The binding and the diffusion of mitoxantrone (MTX) through the plasma membrane was performed by Forster resonance energy transfer (FRET) from the membrane fluorescent donor (4Di-10ASP) to the co-localized acceptor MTX. The MTX addition to living 4Di-10ASP-tagged cells resulted in the rapid quenching of the probe emission (1 s), revealing the MTX binding to the outer leaflet. Then, a slower quenching (about 90 s) occurred which corresponded to the MTX flip-flop into the inner leaflet. Changes of MTX integration into the plasma membrane were described in BCRP-overexpressed cells (HCT-116R) treated with (i) the BCRP inhibitor fumitremorgin C (FTC), (ii) cyclosporin A (CSA) and (iii) benzyl alcohol (BA). Treatments with FTC or CSA showed 80% and 40% higher flip-flop of MTX from the outer to the inner leaflet of HCT-116R cells. The addition of BA clearly increased the MTX integration into both outer and inner leaflets. Confocal fluorescence microscopy displayed that FTC, CSA and BA enhanced MTX accumulation in HCT-116R. In conclusion, Fumitremorgin C and agents modulating MTX accumulation resulted in higher MTX integration in the resistant cell membrane and could disrupt the membrane cohesion. This energy transfer method appears well-adapted to describe the drug diffusion through the plasma membrane of living cells.

International audience; This article describes the quality of reproduction of microcavities structured into mould substrates that were filled by the imposed flow of amorphous polymeric materials. The rheology of the selected materials (cyclic olefin copolymer, COC) depends on the experimental parameters (temperature, pressure) used for the hot embossing process. To support the experimental data, the polymers are qualified by their melt flow index, flow index and consistency. The efficiency of the filling procedure into the microcavities of smaller and smaller size is described using the potentialities of a customized equipment labelled as a Scanning Mechanical Microscopy (SMM).

To study the pH gradient status through membranes of acidic vesicles, either in sensitive or in multidrug-resistant living cancer cells, we monitored the fluorescence-emission spectra of acridine orange. Successive stainings with a pH-sensitive dye and AO showed that low-pH organelles were stained red by AO. In these compartments, high AO concentrations are driven by the pH gradient through membrane vesicles. The resulting rise in the dye's oligomeric/monomeric ratio induced an increase in the red/green (655-nm/530-nm) emission intensity ratio. Therefore, the accumulation of AO in acidic organelles was appraised by determination of the contribution of the red emission intensity (R%) in each emission spectrum, using laser scanning confocal microspectrofluorometry. In vesicles of multidrug-resistant K562-R cells, R% is significantly higher (72 ± 10%) than the value (48 ± 8%) from K562-sensitive cells ( p

Nuclear morphological alterations associated with multidrug resistance (MDR) were evaluated by image cytometry in various human leukemic cell sub-lines: 3 cell lines with P-gp-mediated resistance (CEM-VLB, HL60/Vinc, K562-Dox), the non-Pgp-mediated MDR HL60/AR leukemic cell line with over-expres-sion of MRP, and the at-MDR CEM-VMI leukemic cell line with alteration of topoisomerase II. All these MDR cell sub-lines were obtained by drug selection and were compared with their sensitive counterparts and with the hamster LR73-R cell line obtained by transfection of mouse mdrl cDNA. All MDR cell sub-lines obtained by drug selection displayed decreased DNA Feulgen stainability as compared with their respective sensitive parental cell line, a phenomenon not observed in the trans-fected LR73-R cells. Nuclear texture analysis on G0/G1-selected cell nuclei revealed 2 types of textural phenotype. The first phenotype was characterized by chromatin decondensation with small but compact chromatin clumps, and was observed in drug-selected P-gp-mediated MDR cells (CEM-VLB, HL60-Vinc, K562-Dox) and in the non-P-gp-mediated MDR HL60/AR cell line. The second phenotype was characterized by a condensed and homogeneous chromatin pattern, and was observed in the at-MDR CEM-VMI cell line. LR73-R cells transfected with mdrl cDNA did not display any significant changes in textural pheno-type as compared with sensitive LR73 cells, suggesting that P-gp over-expression alone cannot account for the cytological modi-fications observed in MDR cells. These data suggest that multidrug resistance could be associated with specific nuclear morphological changes which appeared to be a consequence of alterations occurring during selection by cytotoxic drugs rather than of P-gp over-expression. © 1995 Wiley-Liss, Inc.