Éditeur(s) : HAL CCSD Elsevier
Résumé : International audience
ISSN: 0003-4975

hal-01563338
http://hal.univ-brest.fr/hal-01563338
DOI : 10.1016/j.athoracsur.2016.03.093
PUBMED : 27262916

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 Elsevier
Résumé : International audience
Aims: The aims of this study were to compare short-and long-term outcomes for clinical T2N0 oesophageal cancer with analysis of (i) primary surgery (S) versus neoadjuvant therapy plus surgery (NS), (ii) squamous cell carcinoma and adenocarcinoma subsets; and (iii) neoadjuvant chemoradiotherapy versus neoadjuvant chemotherapy.Methods: Data were collected from 30 European centres from 2000 to 2010. Among 2944 included patients, 355 patients (12.1%) had cT2N0 disease; 285 (S) and 70 (NS), were compared in terms of short-and long-term outcomes. Propensity score matching analyses were used to compensate for differences in baseline characteristics.Results: No significant differences between the groups were shown in terms of in hospital morbidity and mortality. Nodal disease was observed in 50% of S-group at the time of surgery, with 20% pN2/N3. Utilisation of neoadjuvant therapy was associated with significant tumour downstaging as reflected by increases in pT0, pN0 and pTNM stage 0 disease, this effect was further enhanced with neoadjuvant chemoradiotherapy. After adjustment on propensity score and confounding factors, for all patients and subset analysis of squamous cell and adenocarcinoma, neoadjuvant therapy had no significant effect upon survival or recurrence (overall, loco-regional, distant or mixed) compared to surgery alone. There were no significant differences between neoadjuvant chemotherapy and chemoradiotherapy in short-or long-term outcomes.Conclusion: The results of this study suggest that a surgery alone treatment approach should be recommended as the primary treatment approach for cT2N0 oesophageal cancer despite 50% of patients having nodal disease at the time of surgery.
ISSN: 0959-8049

hal-01666517
https://hal.univ-lorraine.fr/hal-01666517
DOI : 10.1016/j.ejca.2015.11.024

Éditeur(s) : HAL CCSD CNRS éditions
Résumé : International audience
Die antike Siedlung von Mandeure und Mathay (Epomanduodurum), welche durch ihre Größe und ihre monumentale Ausstattung als die zweitwichtigste Stadt der Sequaner hinter der Civitashauptstadt Besançon betrachtet wird, ist Gegenstand eines interdisziplinarischen Forschungsprogramms. Dieser Artikel stellt eine Bilanz der Ergebnisse der zwischen 2001 und 2004 durchgeführten Forschungen dar. Im Mittelpunkt der Forschungen der ersten Phase stand der zentralen Teil der Siedlung mit dem Bereich Heiligtum-Theater-spätrömische Befestigung, der für die Problematik der Entstehung, der Entwicklung und dem Ende der römischen Besiedlung besonders interessant ist. Verschiedene, sich ergänzende Projekte wurden eingeleitet: Erforschung des alten Dokumentarbestandes, geophysikalische Prospektionen, geoarchäologische Sondierungen, Plangrabungen, architektonische Untersuchunge. Durch diese Projekte war es möglich, die Kenntnisse über die drei wichtigen Großbauten (Heiligtum-Theater-spätrömische Befestigung) grundsätzlich zu vertiefen und insbesondere die geophysikalischen Prospektionsergebnisse lassen in bemerkenswerter Weise einen dichten Baubestand dieses Stadtteils erkennen. Schließlich konnte die Funktion und die Entwicklung der Flußebene des Doubs in diesem Bereich besser verstanden werden als auch die Art und Weise in welcher die antike Monumentalanlage in diesen einzigartigen natürlichen Rahmen einbezogen wurde.
Since 2001, a collective pluridisciplinary research programme is being undertaken on the antique town of Epomanduodurum in the communes of Mandeure and Mathay. This site is considered like the second one of the Sequani, for its dimensions and the monuments, after the civitas capital, Besançon. This article is an assessment of the investigations conducted between 2001 and 2004. During this period, the central part of the site was studied (sanctuary, theatre, Late fortification area), considered as the most significant for the question of the birth, development and decline of the town. Various complementary approaches have been carried out: study of archives, geophysical and geoarchaeological survey, extent excavations, architectural analyses. They lead to a better understanding of the three main buildings (the Celtic and Gallo-Roman sanctuary, the theatre of the Principate, the Late Antique fortification) and they add considerable informations about the pattern of settlement of this area; so it is possible to detect more precisely the spatial organization and finally to understand how the alluvial plain of the Doubs river was functioning and how the monumental complex has been integrated in such a peculiar landscape.
L'agglomération antique d'Epomanduodurum, située sur les communes de Mandeure et Mathay, et considérée comme la seconde du pays séquane par la taille et la parure monumentale, derrière la capitale de cité, Besançon, fait l'objet d'un programme collectif de recherche, pluridisciplinaire, depuis 2001. Cet article dresse un bilan des résultats des fouilles réalisées entre 2001 et 2004. Dans cette première phase, les investigations ont porté principalement sur la partie centrale du site (secteur sanctuaire-théâtrefortification tardive), perçue comme particulièrement intéressante pour la problématique de la naissance, du développement et du déclin de l'agglomération. Différentes approches complémentaires ont été lancées : étude du fond documentaire ancien, prospections géophysiques, sondages géoarchéologiques, fouilles extensives, études architecturales. Elles ont permis d'approfondir la connaissance des trois principaux édifices (sanctuaire laténien et gallo-romain, théâtre du Haut-Empire, fortification du Bas-Empire), de compléter de façon très notable la trame de l'occupation de ce secteur, et donc d'en mieux percevoir l'organisation spatiale, enfin de mieux comprendre le fonctionnement de la plaine alluviale du Doubs et l'insertion dans ce contexte naturel particulier d'un complexe monumental antique.
ISSN: 0016-4119

halshs-00718356
https://halshs.archives-ouvertes.fr/halshs-00718356

Éditeur(s) : HAL CCSD
Résumé : International audience
After defining the High Performance Pre- Computing --referred as HPPC-- concept, the aim of the present study is to develop a prototype whether to approve or not the benefits of this concept. Our application case tries to answer the geophysical issue of coastal flooding. This is an example of an alert system based on the HPPC architecture, thus on pre-computed scenarios. The prototype provides the scientists with an ergonomic and on-demand tool allowing the run of scenarios of any implemented numerical models. These runs are available through a web application which submits the corresponding jobs on the remote french public cluster of HPC@LR. In this study we simulate the waves propagation over a Mediterranean grid using the wave model WaveWatch III⃝R . A reference simulation using usual conditions is approximated using the k-NN algorithm over 12, 98 and then 980 pre-computed scenarios. This simple experiment demonstrates how useful the pre-computing of scenarios is for alert systems as far as enough and relevant scenarios are pre-computed. This is the reason why searches continue in each critical points of the HPPC architecture such as the design of experiment, the approximation of the results by meta-models and the research of the closest scenarios in this big data context.
4th International Conference on Knowledge and Systems Engineering
Danang, Vietnam

lirmm-00736797
https://hal-lirmm.ccsd.cnrs.fr/lirmm-00736797
https://hal-lirmm.ccsd.cnrs.fr/lirmm-00736797/document
https://hal-lirmm.ccsd.cnrs.fr/lirmm-00736797/file/PID2396685.pdf
DOI : 10.1109/KSE.2012.36

Éditeur(s) : HAL CCSD Wiley Open Access
Résumé : Biodiversity continues to decline in the face of increasing anthropogenic pressures such as habitat destruction, exploitation, pollution and introduction of alien species. Existing global databases of species' threat status or population time series are dominated by charismatic species. The collation of datasets with broad taxonomic and biogeographic extents, and that support computation of a range of biodiversity indicators, is necessary to enable better understanding of historical declines and to project - and avert - future declines. We describe and assess a new database of more than 1.6 million samples from 78 countries representing over 28,000 species, collated from existing spatial comparisons of local-scale biodiversity exposed to different intensities and types of anthropogenic pressures, from terrestrial sites around the world. The database contains measurements taken in 208 (of 814) ecoregions, 13 (of 14) biomes, 25 (of 35) biodiversity hotspots and 16 (of 17) megadiverse countries. The database contains more than 1% of the total number of all species described, and more than 1% of the described species within many taxonomic groups - including flowering plants, gymnosperms, birds, mammals, reptiles, amphibians, beetles, lepidopterans and hymenopterans. The dataset, which is still being added to, is therefore already considerably larger and more representative than those used by previous quantitative models of biodiversity trends and responses. The database is being assembled as part of the PREDICTS project (Projecting Responses of Ecological Diversity In Changing Terrestrial Systems - ). We make site-level summary data available alongside this article. The full database will be publicly available in 2015.
ISSN: 2045-7758

hal-01204216
https://hal.archives-ouvertes.fr/hal-01204216
DOI : 10.1002/ece3.1303
PRODINRA : 280886