Éditeur(s) : HAL CCSD
Résumé : International audience
STUDY QUESTION: Do freezing and in vitro culture procedures enhance the expression of proteins involved in apoptotic or autophagic pathways in murine pre-pubertal testicular tissue? SUMMARY ANSWER: IVM strongly modified apoptosis- and autophagy-related relative protein levels in mice testicular tissue whereas the impact of cryopreservation procedures was minimal at the end of the culture. WHAT IS KNOWN ALREADY: In vitro spermatogenesis remains a challenging technical issue as it imposes to find a very close balance between survival and death of germ cell natural precursors (i.e. gonocytes and spermatogonia), which will eventually undergo a complete spermatogenesis close to in vivo conditions. The establishment of efficient culture conditions coupled with suitable cryopreservation procedures (e.g. controlled slow freezing [CSF] and solid surface vitrification [SSV]) of pre-pubertal testicular tissue is a crucial step in the fields of fertility preservation and restoration to improve the spermatic yield obtained in vitro. STUDY DESIGN, SIZE, DURATION: Here, we study cryopreservation procedures (i.e. CSF or SSV) and the impact of culture media compositions. A first set of 66 mouse pre-pubertal testes were directly cultured during 30, 36, 38 and 60 days (D) from 2.5 to 6.5-day-old CD-1 mice to evaluate the impact of time-aspect of culture and to endorse the reverse phase protein microarrays (RPPM) technique as an adapted experimental tool for the field of in vitro spermatogenesis. Ninety others fresh, slow-frozen and vitrified pre-pubertal testes were cultured during 30 days for the principal study to evaluate the impact of cryopreservation procedures before and after culture. Thirty-four testes dissected from 2.5, 6.5, 36.5, 40.5, 42.5 and 62.5 days postpartum (dpp) mice, corresponding to the time frames of spermatogenesis orchestrated in vitro, were used as in vivo controls. PARTICIPANTS/MATERIALS, SETTING, METHODS: After in vitro culture, testicular tissue samples originated from 2.5 or 6.5-day-old CD-1 male mice were analyzed using RPPM. This targeted proteomic technique allowed us to assess the expression level of 29 apoptosis- and autophagy-related factors by normalizing blank-corrected signal values. In addition, morphological analyses (e.g. HES, PAS, TRA98 and CREM) and DNA fragmentation in intra-tubular cells (i.e. terminal deoxynucleotidyl transferase dUTP nick end labeling; TUNEL) were assessed for the distinct experimental conditions tested as well as for in vivo control mouse testes. MAIN RESULTS AND THE ROLE OF CHANCE: A validation of the RPPM procedure in the field of in vitro spermatogenesis was completed with assay and array robustness before a principal study concerning the evaluation of the impact of in vitro culture and cryopreservation procedures. The proportion of elongated spermatids and the total cell number per seminiferous tubule tended to be very different between the in vivo and in vitro conditions (P < 0.05), suggesting the presence of a beneficial regulation on the first spermatogenesis wave by intrinsic apoptosis (Caspase₉) and autophagy (Atg5) factors (P < 0.0003 and r2 = 0.74). Concerning the impact of culture media compositions, a basic medium (BM) composed of αMEM plus 10% KnockOut™ serum replacement and gentamicin supplemented with retinol (Rol) and vitamin E (Vit. E) was selected as the best culture medium for fresh 6.5 dpp tissue cultured during 30D with 27.7 ± 8.10% of seminiferous tubules containing elongated spermatids. Concerning the impact of cryopreservation procedures, SSV did not have any impact on the morphological parameters evaluated after culture in comparison to fresh tissue (FT) controls. The proportion of tubules with elongated spermatids on testicular explants cultured with BMRol+Vit. E was not different between SSV (6.6 ± 1.6%) and CSF (5.3 ± 1.9%); however, round spermatids were observed more frequently for SSV (19 ± 6.2%) than CSF (3.3 ± 1.9%, P = 0.0317). Even if the proportion of TUNEL-positive cells for BMRol+Vit. E was higher at D30 after SSV (4.12 ± 0.26%) than CSF (1.86 ± 0.12%, P = 0.0022) and FT (2.69 ± 0.33%, P = 0.0108), the DNA damages observed at the end of the culture (i.e. D30) were similar to respective 6.5 dpp controls. In addition, the relative protein level expression ratio of an apoptotic factor, the phosphorylated FADD on Fas, was reduced by 64-fold in vitrified testes cultured with BMRol+Vit. E. Furthermore, we found in this study that the StemPro®-34 SFM culture medium supplemented with growth factors (e.g. EGF, bFGF, GDNF and LIF) prevented the differentiation of spermatogonial stem cells in favor of a significant proliferation with a better architectural pattern than in vivo 6.5 dpp controls with an increase of seminiferous tubules area for FT (P = 0.0357) and CSF (P = 0.0317). LIMITATIONS REASONS FOR CAUTION: Despite our promising results, the evaluation of apoptotic- and autophagic-related proteins was studied for a limited amount of proteins and on global testicular tissue. WIDER IMPLICATIONS OF THE FINDINGS: The data presented herein will help to improve apoptotic and autophagic understanding during the first spermatogenic wave. Moreover, our findings illustrate for the first time that, using finely-tuned experimental conditions, a testicular in vitro culture combined with proteomic technologies may significantly facilitate the study of cryopreservation procedures and in vitro culture evaluations. This study may also contribute to improve work on testicular tissues from pre-pubertal and adolescent cancer survivors. STUDY FUNDING/COMPETING INTEREST(S): This study was supported by a Ph.D. grant from the Rouen Normandie Université and a financial support from 'la Ligue nationale contre le cancer' (both awarded to L.D.), funding from Institute for Research and Innovation in Biomedicine (IRIB), Agence de la Biomédecine, and co-supported by European Union and Région Normandie. Europe gets involved in Normandie with European Regional Development Fund (ERDF). The authors declare that there is no conflict of interest.
Molecular Human Reproduction

hal-01647081
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01647081
DOI : 10.1093/molehr/gax054
PUBMED : 29040674

Éditeur(s) : HAL CCSD Springer/Terra Scientific Publishing Company
Résumé : International audience
We report oil the discovery of magnetite nanoparticles ranging in size from 2 to 14 full in the mineralized zones of the Pena Colorada iron-ore deposit, southern Mexico. Micrometric scale magnetite was magnetically reduced and divided into distinct size ranges: 85-56 mu m, 56-30 mu m, 30-22 mu m, 22-15 mu m, 15-10 mu m, 10-7 mu m and 7-2 mu m. Nanometric-scale magnetite in the size range 2-14 nm was identified. The magnetite was characterized by X-ray diffraction, transmitted and reflected light microscope, high-resolution transmission electron microscopy (TEM), high angle annular dark field, Mossbauer spectroscopy and its magnetic properties. Crystallographic identification of nanostructures Was performed using high-resolution TEM. Characteristic changes were observed when the particles make the size transition from micro- to nanometric sizes, as follows: (1) frequency-dependent magnetic Susceptibility percentage (chi FD%) measurements Show high Values (13%) for the 2-14 nm fractions attributed to dominant fractions of superparamagnetic particles; (2) variations of chi FD% < 4.5% in fractions of 56-0.2 mu m occur in association with the presence of microparticles formed by magnetite aggregates of nanoparticles (< 15 nm) embedded in berthierine; (3) Mossbauer spectroscopy results identified a superparamagnetic fraction; (4) nanometric and 0.2-3 mu m grain size magnetite particles require a magnetic field up to 152 mT to reach saturation during the isothermal remanent magnetization experiment; (5) coercivity and remanent magnetization of the magnetite increase when the particle size decreases, probably due to parallel Coupling effects; (6) two-magnetic Susceptibility versus temperature experiments of the same 2-14 nm sample show that the reversibility during the second heating is due to the formation of new magnetite nanoparticles and growth of those already present. during the first heating process.
ISSN: 1343-8832

hal-00413053
https://hal.archives-ouvertes.fr/hal-00413053

Éditeur(s) : HAL CCSD Springer Verlag (Germany) Springer Verlag
Résumé : Direct numerical simulations of flow through two millimeter-scale rock samples of limestone and sandstone are performed using three diverse fluid dynamic simulators. The resulting steady-state velocity fields are compared in terms of the associated empirical probability density functions (PDFs) and key statistics of the velocity fields. The pore space geometry of each sample is imaged at 5.06−μm voxel size resolution using X-ray microtomography. The samples offer contrasting characteristics in terms of total connected porosity (about 0.31 for the limestone and 0.07 for the sandstone) and are typical of several applications in hydrogeology and petroleum engineering. The three-dimensional fluid velocity fields within the explicit pore spaces are simulated using ANSYS® FLUENT® ANSYS Inc. (2009), EULAG Prusa et al. (Comput. Fluids 37, 1193–1207 2008), and SSTOKES Sarkar et al. (2002). These computational approaches are highly disperse in terms of algorithmic complexity, differ in terms of their governing equations, the adopted numerical methodologies, the enforcement of internal no-slip boundary conditions at the fluid-solid interface, and the computational mesh structure. As metrics of comparison to probe in a statistical sense the internal similarities/differences across sample populations of velocities obtained through the computational systems, we consider (i) integral quantities, such as the Darcy flux and (ii) main statistical moments of local velocity distributions including local correlations between velocity fields. Comparison of simulation results indicates that mutually consistent estimates of the state of flow are obtained in the analyzed samples of natural pore spaces despite the considerable differences associated with the three computational approaches. We note that in the higher porosity limestone sample, the structures of the velocity fields obtained using ANSYS FLUENT and EULAG are more alike than either compared against the results obtained using SSTOKES. In the low-porosity sample, the structures of the velocity fields obtained by EULAG and SSTOKES are more similar than either is to the fields obtained using ANSYS FLUENT. With respect to macroscopic quantities, ANSYS FLUENT and SSTOKES provide similar results in terms of the average vertical velocity for both of the complex microscale geometries considered, while EULAG tends to render the largest velocity values. The influence of the pore space structure on fluid velocity field characteristics is also discussed.
ISSN: 1420-0597

hal-01174152
https://hal.archives-ouvertes.fr/hal-01174152
DOI : 10.1007/s10596-015-9486-7

Éditeur(s) : HAL CCSD Springer/Terra Scientific Publishing Company
Résumé : International audience
This study of the Cajati deposit provides evidence that the ore was neither purely hydrothermal, nor volcanic in origin, as previous workers have proposed. The ores were formed from magnetite-rich magmas, hydrothermally altered and intruded at an indicated crustal depth in excess of 500 m. The mineralogical and textural association between magnetite and magnesioferrite in the carbonatite, and between the titanomagnetite and magnesioferrite-Ti mineralization in the pyroxenite of hedenbergite, seems to be analog mineralizations strongly related to the ionic substitution of Fe2+ by Mg. Relatively high Q ratios (>= 5) for Jacupirangite-pyroxenite may indicate a thermo remanent magnetization (TRM) by the ore during post-metamorphic cooling, however it call also be developed from chemical remanent magnetization (CRM). Vector plots for the pyroxenite samples show reasonably linear and stable magnetic components. The intensity decay Curves show that only two components of magnetizations are likely present. Continuous susceptibility measurements with increasing temperature show that the main magnetic phase seems to be magnetite. Maghemite is probably produced during the cooling process. Susceptibility recorded from low temperature (liquid nitrogen (- 196 degrees C)) to room temperature produces typical Curves. indicating Verwey transition of magnetite. Hysteresis parameters point Out that nearly all values fill in a novel region of the Day plot, parallel to but below magnetite SD + MD mixing curves.
ISSN: 1343-8832

hal-00420922
https://hal.archives-ouvertes.fr/hal-00420922

Éditeur(s) : HAL CCSD Wiley
Résumé : International audience
ISSN: 0933-7407

hal-01632369
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01632369

Éditeur(s) : HAL CCSD Springer Verlag (Germany) Springer Verlag
Résumé : International audience
The dissolution of carbonate rocks usually leads to both porosity (ϕ) and permeability (k) increase. We present experimental evidences and physical-based models of positive and anti-correlated dynamics of k and ϕ observed during dissolution experiments of carbonate rocks. We study the way the rate of change of ϕ and k is controlled by the degree of undersaturation of the percolating solution for two different types of carbonate rocks. We document the occurrence of an anti-correlated k−ϕ trend when the flowing fluid (deionized water) has a weak capacity of dissolution. A positive correlation is found when CO2 is added to the deionized water to increase the potential dissolution rate. Detailed analyses of the microstructures of the rock performed by X-ray microtomography reveal that low dissolution rate favors detachment of solid particles and their subsequent accumulation at the pore-throat inlet. Particles are detached from the rock matrix due to the differential dissolution rate of the indurated grains and the microporous cement. We then propose a simple phenomenological model to interpret the effect of the pore-throat clogging by the accumulation of partially dissolved carbonate particles. We conjecture that permeability is controlled by the decrease of the effective hydraulic radius and the increase of the tortuosity due to partial and localized obstruction of the pore network. Conversely, increasing the level of undersaturation of the flowing solution leads to an augmented potential of dissolving most of the transported particles before they reach the throats. In this case, both k and ϕ increase and display power-law correlations.
ISSN: 0169-3913

hal-01174145
https://hal.archives-ouvertes.fr/hal-01174145
DOI : 10.1007/s11242-015-0456-2

Éditeur(s) : HAL CCSD Elsevier Masson
Résumé : National audience
Introduction L’objectif de cette étude est de disposer de données qualitatives et quantitatives sur la contamination de l’environnement intérieur hospitalier, microbiologique et chimique, afin d’évaluer l’exposition du personnel, des visiteurs et des patients et la variabilité spatio-temporelle de la contamination. Méthodes L’étude s’est déroulée dans différents lieux des CHU de Rennes et de Nancy : hall d’accueil, salle de soins infirmiers, salle de réveil post-opératoire, chambre d’un patient, unité de désinfection des endoscopes, laboratoire de parasitologie et salle de découpe de plâtres. Deux campagnes de prélèvement ont eu lieu en été 2014 et en hiver 2015. Les méthodes de prélèvement étaient nombreuses et complémentaires (Fig. 1). L’analyse des données a été réalisée sous IBM SPSS grâce au test de Mann-Whitney et le coefficient de corrélation de Pearson. Résultats Les concentrations d’aldéhydes, de composés organiques volatils (COV) et semi-volatils étaient faibles à très faibles dans les 2 établissements. Étaient détectables, mais sans différences globales entre les 2 établissements : la contamination fongique (m = 226 UFC/m3 ; p = 0,97) ou bactérienne (m = 352 UFC/m3 ; p = 0,14), les particules PM2,5 (m = 2,1 μg/m3 ; p = 0,97) et PM10 (m = 6,6 μg/m3 ; p = 0,84). Sur le plan saisonnier, la contamination fongique était plus importante en été pour les 2 établissements (p = 0,002). La contamination bactérienne ne varie pas significativement entre les locaux (p = 0,14) contrairement à la contamination fongique (p = 0,02) et particulaires PM2,5 (p = 0,01) ou PM10 (p = 0,01). Cette dernière est plus importante dans le hall (m = 879 UFC/m3), le laboratoire de parasitologie (m = 333 UFC/m3) et la salle de plâtre (m = 310 UFC/m3). Nous ne retrouvons pas de corrélation significative entre le nombre de personnes, le taux de CO2, l’humidité relative et le dénombrement bactérien ou fongique. Néanmoins nous retrouvons une relation forte entre la température du local et la contamination bactérienne (r = 0,7, p = 0,008) ou la contamination fongique (r = 0,56, p = 0,045) d’une part, et entre la contamination fongique et les PM10 (r = 0,52, p = 0,022) d’autre part. Conclusion Notre étude montre une très faible contamination par les aldéhydes, COV et sCOV comparativement à d’autres lieux publics. La forte variabilité spatio-temporelle de la contamination fongique et particulaire est liée à la saison, à l’activité et à la ventilation
ISSN: 1156-5233

hal-01334070
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01334070
DOI : 10.1016/j.mycmed.2016.04.026

Éditeur(s) : HAL CCSD
Résumé : International audience
OBJECTIVE: To assess sperm production and aneuploidy in Hodgkin lymphoma (HL) and non-Hodgkin lymphoma (NHL) before and after treatments. DESIGN: Multicenter, prospective, longitudinal study of lymphoma patients analyzed before treatment and after 3, 6, 12, and 24 months. SETTING: University hospitals. PATIENT(S): Forty-five HL and 13 NHL patients were investigated before and after treatment. Treatment regimens were classified in two groups: ABVD (doxorubicin, bleomycin, vinblastine, dacarbazine) with or without (+/-) radiotherapy, and CHOP (doxorubicin, cyclophosphamide, vincristine, prednisone)/MOPP-ABV (mechlorethamine, oncovin, procarbazine, prednisone-doxorubicin, bleomycin, vinblastine). A control group of 29 healthy men was also studied. INTERVENTION(S): Semen analyses and aneuploidy study by FISH were performed at each time point. MAIN OUTCOME MEASURE(S): Comparison of mean sperm characteristics and percentage of sperm aneuploidy rates before and after treatment. RESULT(S): Before treatment, HL and NHL men had altered semen characteristics and higher sperm aneuploidy rates (median 0.76 [interquartile range 0.56-0.64]) than the control group (0.54 [0.46-0.74]). After treatment, sperm production was significantly lowered 3 and 6 months after ABVD +/- radiotherapy or CHOP/MOPP-ABV. After ABVD +/- radiotherapy, the aneuploidy rate increased significantly only at 3 months, and values obtained 1 or 2 years later were lower than pretreatment values. In contrast, in the CHOP/MOPP-ABV treatment group, semen characteristics and aneuploidy rate did not return to normal levels until 2 years after treatment. CONCLUSION(S): Lymphoma itself has consequences on sperm aneuploidy frequency before treatment. Moreover, lymphoma treatments have deleterious effects on sperm chromosomes related to treatment type and time since treatment. Patient counseling is essential concerning the transient but significant sperm aneuploidy induced by lymphoma and its treatments.
Fertility and sterility

hal-01477204
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01477204
DOI : 10.1016/j.fertnstert.2016.10.001
PUBMED : 27810161

Éditeur(s) : HAL CCSD Geological Society
Résumé : Extensional basins often evolve both with the generation of normal faults at all scales, and with tilting in the hanging walls and footwalls of major structures. This paper examines the effect of regional tilting on the development of normal fault arrays in an anisotropic sequence, using the asymmetric Lodève Basin, France, as a case study. The orientation range of normal faults is found not to match that expected by continued symmetric generation of conjugate normal faults during tilting. Instead, continued generation of new faults preferentially occurred for faults dipping opposite to the direction of tilt, with propagation perpendicular to bedding. The presence of early bedding-perpendicular joints in the competent sandstone units aided propagation of these faults during tilting and this preferential asymmetric development. Bedding-parallel slip surfaces were activated as normal faults during the later stages of bedding tilting, showing complex interaction patterns with later faults downthrowing in the direction opposite to bedding tilting. Hence knowledge of the timing between tilting of the strata and deformation helps in evaluating the likely complexity of fault arrays at subseismic scales in extensional basins, and may also help in understanding the evolution of fault systems in tilted normal fault blocks at the reservoir scale.
ISSN: 0016-7649

hal-00404753
https://hal.archives-ouvertes.fr/hal-00404753
DOI : 10.1144/0016-76492006-047