Fonds National De La Recherche Research Articles

Influence of follicle stage on artificial ovary outcome using fibrin as a matrix.

Do primordial-primary versus secondary follicles embedded inside a fibrin matrix have different capabilities to survive and grow after isolation and transplantation? Mouse primordial-primary follicles showed a lower recovery rate than secondary follicles, but both were able to grow. Fresh isolated mouse follicles and ovarian stromal cells embedded in a fibrin matrix are capable of surviving and developing after short-term autografting. In vivo experimental model using 11 donor Naval Medical Research Institute (NMRI) mice and 11 recipient severe combined immunodeficiency (SCID) mice. Both ovaries from all NMRI mice were mechanically disrupted and primordial-primary and secondary follicles were isolated with ovarian stromal cells. They were then encapsulated in a fibrin matrix composed of 12.5 mg/ml of fibrinogen (F12.5) and 1 IU/ml of thrombin (T1) (F12.5/T1), and grafted to the inner part of the peritoneum of SCID mice for 2 and 7 days. This study was conducted at the Gynecology Research Unit, Université Catholique de Louvain. All materials were used to conduct histological (H-E staining) and immunohistochemical (Ki67, TUNEL) analyses. Although all grafted fibrin clots were recovered, the follicle recovery rate on day 2 was 16 and 40% for primordial-primary and secondary follicles respectively, while on day 7, it was 6 and 28%. The secondary group showed a significantly higher recovery rate than the primordial-primary group (23%, P-value <0.001). Follicles found in both groups were viable, as demonstrated by live/dead assays, and no difference was observed in the apoptosis rate between groups, as evidenced by TUNEL. Their growth to further stages was confirmed by Ki67 immunostaining. As demonstrated by our results, secondary follicles appear to be more likely to survive and develop than primordial-primary follicles in a fibrin matrix after both periods of grafting. These findings may also be attributed to the specific features of the fibrin matrix, which could benefit larger follicles, but not smaller follicles. This study is essential to understanding possible impairment caused by factors such as the isolation procedure or fibrin matrix composition to the survival and development of different follicle stages. It therefore provides the basis for further investigations with longer periods of grafting. This study was supported by grants from the Fonds National de la Recherche Scientifique de Belgique (grant Télévie No. 7.4578.14 and 7.4627.13, grant 5/4/150/5 awarded to Marie-Madeleine Dolmans), Fonds Spéciaux de Recherche, Fondation St Luc, the Foundation Against Cancer, and the Region Wallone (Convention N°6519-OVART) and donations from Mr Pietro Ferrero, Baron Frère and Viscount Philippe de Spoelberch. None of the authors have any competing interests to declare.

The best source of isolated stromal cells for the artificial ovary: medulla or cortex, cryopreserved or fresh?

What is the best source of ovarian cells for the artificial ovary: medulla or cortex, cryopreserved or fresh? Ovarian cells from fresh medullary tissue, which can be isolated in larger numbers, show higher viability and are able to improve graft vascularization. In a previous study, addition of endothelial cells along with ovarian cells was found to be crucial for formation of a well-vascularized ovary-like structure. This study is the first to evaluate both the effect of cryopreservation and the source of ovarian tissue on isolated ovarian cells. Prospective experimental study in an academic research unit using ovarian tissue from seven patients undergoing surgery for benign gynecologic disease. Ovarian tissue was retrieved from seven patients, with one half processed as fresh (fresh group) and the other half frozen and thawed before processing (frozen group). In each group, ovarian cells from the cortex and medulla were isolated separately, and their viability was tested using a calcein AM/ethidium homodimer viability assay. Fifty thousand cells were then encapsulated in fibrin and grafted to peritoneal pockets in nude mice (14 in all). Grafts recovered after 7 days were analyzed by immunohistochemistry for the presence of ovarian cells (vimentin), proliferation (Ki67) and graft vascularization (double CD34). Cell apoptosis was analyzed by TUNEL assay. Cryopreservation decreased ovarian cell yield (-2804 cells/mg, P = 0.015) and viability (-9.72%, P = 0.052) before grafting and had a considerable (5-fold, P = 0.2) but non-significant negative impact on ovarian cell presence in grafts. The medulla yielded many more cells (+3841 cells/mg, P < 0.001) with higher viability (+18.23%, P < 0.001) than did the cortex. Moreover, grafts with cells from the medulla exhibited a statistically significant 6.44- and 2.47-fold increase in human and total vascular surface area, respectively. P-values were adjusted for multiple testing using the Benjamini-Hochberg method to achieve a 10% false discovery rate and adjusted P-values < 0.1 were therefore considered significant. Pilot study involving a limited number of experiments. Knowing that fresh medullary tissue is the best source of stromal cells is important for construction of the artificial ovary, as isolated follicles require structural support and a rich vascular network for their survival and development. This work was supported by grants from the Fonds National de la Recherche Scientifique de Belgique (5/4/150/5 and 7.4518.12F), Fonds Spéciaux de Recherche, Fondation Saint Luc and Foundation Against Cancer, and donations from Mr Pietro Ferrero, Baron Frère and Viscount Philippe de Spoelberch. None of the authors have any conflicting interests to declare.

Switching Reconstruction of Digraphs

Australian Research Council; contract grant number: DP1093320; contract grant sponsor: Fonds National de la Recherche, Luxembourg, co-funded under the Marie Curie Actions of the European Commission; contract grant number: FP7-COFUND.

Vitrification preserves proliferation capacity in human spermatogonia

Does vitrification of human immature testicular tissue (ITT) have potential benefits for future fertility preservation? Does vitrification of human ITT have potential benefits in an in vivo murine xenotransplantation model? Vitrification is able to maintain proliferation capacity in spermatogonial cells after 6 months of xenografting. Controlled slow-freezing is the procedure currently applied for ITT cryobanking in clinical practice. Vitrification has been proposed as a promising technique for long-term storage of ITT, with a view to preserving spermatogonial stem cells (SSCs) for future fertility restoration in young boys suffering from cancer. After vitrification of ITT, in vitro survival of SSCs was demonstrated, but their functionality was not evaluated. Ten ITT pieces issuing from 10 patients aged 2-12 years were used. Fragments of fresh tissue (serving as controls) and fresh, frozen-thawed and vitrified-warmed testicular pieces xenografted to the scrotum of nude mice for 6 months were compared. Upon graft removal, histological and immunohistochemical analyses were performed to evaluate spermatogonia (SG) (MAGE-A4), intratubular proliferation (Ki67), proliferating SG and Leydig cells (3β-HSD). The entire piece of grafted tissue was assessed in each case. Seminiferous tubules showed good integrity after cryopreservation and xenografting for 6 months in all three groups. Survival of SG and their ability to proliferate was observed by immunohistochemistry in all grafted groups. SG were able to initiate spermatogenesis, but blockage at the pachytene stage was observed. The recovery rate of SG was 3.4 ± 3.8, 4.1 ± 7.3 and 7.3 ± 6.3%, respectively, for fresh, slow-frozen and vitrified-warmed tissue after 6 months of xenografting. The study is limited by the low availability of ITT samples of human origin. The mouse xenotransplantation model needs to be refined to study human spermatogenesis. The findings of the present study have potential implications for cryobanking of ITT and fertility preservation. Spermatogonial loss recorded after fresh ITT transplantation indicates that the avascular grafting technique needs to be optimized. There are so far no convincing data justifying modification of current clinical practice for ITT storage with slow-freezing, but this study demonstrates that it is worth pursuing optimization of ITT vitrification as an alternative for preservation of SSCs. The present study was supported by a grant from the Fonds National de la Recherche Scientifique de Belgique (grant Télévie N° 7. 4.572.09.F). The authors declare that there is no conflict of interest.

256 HOXB9 KNOCK-DOWN DURING OOCYTE MATURATION IN CATTLE

HOXB9 is a transcription factor playing a major role in patterning the main body-axis of vertebrate embryos from the gastrulation stage. HOXB9 transcripts have been detected in cattle oocytes and early embryos (Paul et al. 2011 Mol. Reprod. Dev. 78, 436–449), but their role at those stages has remained unknown. The aim of the study was to inject small interfering RNA (siRNA) in immature oocytes to investigate HOXB9 functions during oocyte maturation and early development. Immature cow oocytes were injected with 7 pL of a HOXB9 siRNA mixture (Integrated DNA Technologies, Coralville, IA, USA) or with a negative control siRNA (SCR) or were not injected (control). To allow microinjection, the size of the cumulus was reduced by pipetting; however, the corona radiata was left intact. The efficiency of the knock-down was measured 24 h post-injection using RT-qPCR, and the results were normalized with an internal normalization factor corresponding to the mean expression of three housekeeping genes (Goossens et al. 2005 BMC Dev. Biol. 5, 27). The injection of HOXB9 siRNA induced a 60.7% reduction of HOXB9 relative expression in mature oocytes by comparison with the injection of SCR siRNA (Kruskall-Wallis; P &lt; 0.05). The nuclear maturation rates, assessed 24 h post-injection after Hoechst staining, were not statistically different between the HOXB9 group (48.8%) and the SCR group (53.6%; chi-square; P &gt; 0.05). After in vitro fertilization and culture (Paul et al. 2011 Mol. Reprod. Dev. 78, 436–449), the developmental rates of the HOXB9 and the SCR groups were not statistically different in terms of achieving the first cleavage (64.4% v. 57.0%, respectively) and 5- to 8-cell stages (25.1% v. 25.3%, respectively) at 48 h post-insemination and the blastocyst stage on Day 7 (IVF = Day 0; 22.0% v. 20.6%, respectively) and Day 8 (26.9% v. 23.1%, respectively; two-way ANOVA; P &gt; 0.05). Differential staining of Day 8 blastocysts showed no difference between the HOXB9 and the SCR groups regarding the mean number of cells in the inner cell mass (53.9 v. 49.4, respectively), the trophectoderm (125.5 v. 104.3, respectively), or the total number of cells (two-way ANOVA; P &gt; 0.05). However, the microinjection procedure impaired the developmental competence after fertilization of the injected oocytes (HOXB9 + SCR) compared with the non-injected ones, as shown by their lower ability to cleave (60.7% v. 72.1%, respectively), to reach the 5- to 8-cell stage (25.2% v. 35.1%, respectively) or the blastocyst stage on Day 7 (21.3% v. 29.8%, respectively) and Day 8 (25.0% v. 35.8%, respectively; two-way ANOVA; P &lt; 0.05). The injection did not affect blastocyst cell numbers on Day 8 (two-way ANOVA; P &gt; 0.05). In conclusion, microinjection of siRNA in immature oocytes is a valuable method to efficiently knock down the maternal messenger RNA during oocyte maturation. Indeed, HOXB9 knock-down in immature oocytes led to a significant and important reduction in the amount of transcript in mature oocytes. The absence of effect of the knock-down on nuclear maturation and embryo development could be explained by the persistence of the maternal proteins. This work was funded by the Fonds National de la Recherche Scientifique (Belgium).

213 HOXB9 PROTEIN IS PRESENT THROUGHOUT EARLY EMBRYO DEVELOPMENT IN THE BOVINE

Hox genes encode for homeodomain transcription factors well known to be involved in developmental control after gastrulation. However, the expression of some of these genes has been detected during oocyte maturation and early embryo development. An interesting expression profile has been obtained for HOXB9 in the bovine (Paul et al. 2011 Mol. Reprod. Dev. 78, 436): its relative expression increases between the immature oocyte and the zygote, further increases at the 5- to 8-cell stage to peak at the morula stage before decreasing at the blastocyst stage. The main objective of this work is to establish the HOXB9 protein profile from the immature oocyte to the blastocyst in the bovine. Bovine embryos were produced in vitro from immature oocytes obtained from slaughterhouse ovaries. Embryos were collected at the following stages: immature oocyte, mature oocyte, zygote (18 h post-insemination, hpi), 2-cell (26 hpi), 5 to 8 cell (48 hpi), 9 to 16 cell (96 hpi), morula (120 hpi), and blastocyst (180 hpi). The presence and distribution of HOXB9 proteins were detected by whole-mount immunofluorescence followed by confocal microscopy using an anti-human HOXB9 polyclonal antibody directed against a sequence showing 100% homology with the bovine protein. Its specificity to the bovine protein was controlled by Western blot on total protein extract from the bovine uterus and revealed, among a few bands of weak intensities, 2 bands of high intensity corresponding to the expected size. Oocytes or embryos were fixed and incubated overnight with rabbit anti-HOXB9 (Sigma, St. Louis, MO, USA) and mouse anti-E-cadherin (BD Biosciences, Franklin Lakes, NJ, USA) primary antibodies and then for 1 h with goat anti-rabbit Alexafluor 555 conjugated (Cell Signaling Technology, Beverly, MA, USA) and goat anti-mouse FITC-conjugated (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA) secondary antibodies. Embryos were then mounted in Vectashield containing DAPI. HOXB9 is detected from the immature oocyte to the blastocyst stage. At the immature oocyte stage, it is mainly localised in the germinal vesicle with a weak signal in the cytoplasm. At the mature oocyte stage, HOXB9 labelling is present in the cytoplasm. At the zygote stage, a stronger immunoreactivity is observed in the pronuclei than in the cytoplasm. From the 2-cell stage to the morula stage, the presence of HOXB9 is also more important in the nuclei than in the cytoplasm. HOXB9 is also observed at the blastocyst stage where it is localised in the nuclei of the trophectoderm cells, whereas an inconstant or weaker labelling is observed in the inner cell mass cells. In conclusion, we have shown for the first time the presence of the HOXB9 protein throughout early bovine embryo development. The results obtained suggest the presence of the maternal HOXB9 protein because it is already detected before the maternal to embryonic transition that occurs during the fourth cell cycle in the bovine. Finally, the pattern obtained at the blastocyst stage suggests a differential role of HOXB9 in the inner cell mass and trophectoderm cells. C. Sauvegarde holds a FRIA PhD grant from the Fonds National de la Recherche Scientifique (Belgium).

More efficient ways of assessing treatments for neglected tropical diseases are required: innovative study designs, new endpoints, and markers of effects.

The authors have declared that no competing interests exist. The Fonds National de la Recherche of Luxembourg provided funds to support the submission of the paper (FNR/11/AM4/55, http://www.fnr.lu). Information on the funding of the study referred to is provided in Sundar et al., CID 2008 Oct 15;47(8):1000-6. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Nociception Coma Scale-Revised total scores correlate with anterior cingulate cortex as measured by FDG-PET in patients with disorders of consciousness

Event Abstract Back to Event Nociception Coma Scale-Revised total scores correlate with anterior cingulate cortex as measured by FDG-PET in patients with disorders of consciousness Camille Chatelle1*, Aurore Thibaut1, Marie-Aurélie Bruno1, Melanie Boly1, Claire Bernard2, Roland Hustinx2, Caroline Schnakers1 and Steven Laureys1 1 University of Liège, Cyclotron Research Centre, Belgium 2 University Hospital of Liège, Nuclear Medicine Dept, Belgium Introduction. The Nociception Coma Scale-Revised was recently validated to assess possible pain perception in patients with disorders of consciousness (i.e., vegetative state/unresponsive wakefulness syndrome and minimally conscious state [1,2,3]). We here aim to identify correlations between cerebral glucose metabolism and Nociception Coma Scale-Revised total scores in disorders of consciousness. Methods. [18F]-fluorodeoxyglucose-PET was performed in 42 patients with disorders of consciousness (29 males, aged 42±17 years; 26 minimally conscious state; 19 post-traumatic; 22 in the acute/subacute setting (i.e., < 1 year post insult). All patients had Coma Recovery Scale-Revised [4] and Nociception Coma Scale-Revised assessment on the day of PET imaging. Data preprocessing and analyses employed statistical parametric mapping (SPM8) and results were considered significant at small-volume corrected p<0.05 for a priori identified coordinates of the pain matrix (Boly et al., 2008 [5]) (i.e., thalamus, primary and secondary somatosensory cortex, insula and anterior cingulate cortex). Results. Anterior cingulate cortex metabolism showed a positive linear correlation with Nociception Coma Scale-Revised total scores and not with Coma Recovery Scale-Revised total scores (corrected p= 0.018; x=20, y=16, z=36). The observed effect showed no significant difference depending on etiology or duration. Conclusions. Nociception Coma Scale-Revised scores (and not Coma Recovery Scale-Revised scores) correlated with metabolic activity of anterior cingulate cortex, known to be involved in the affective emotional component of pain perception. Our data support the hypothesis that Nociception Coma Scale-Revised assessment in non-communicative patients with disorders of consciousness indeed reflects possible pain perception, permitting clinicians to treat adequately and optimize the clinical management of this challenging patient population [6]. Figure 1 Acknowledgements CC is Research Fellow; MAB, MB and CS are Postdoctoral Researcher and SL is Research Director at the Fonds National de la Recherche Scientifique de Belgique (FNRS). This research was supported by FNRS, University and University Hospital of Liège, European Commission, James S. McDonnell Foundation, Mind Science Foundation, Concerted Research Action and Fondation Léon Frédéricq.

Conference Scene: Omic Technologies in Human Disease: Extending the Network of Epigenetic Control

The meeting 'Cell signal-omics 2011: integrated cellular pathology - systems biology of human disease' held on 26-28 January 2011 in Luxembourg, was organized by Recherches Scientifiques Luxembourg (RSL) under the auspices of the European Research Institute for Integrated Cellular Pathology. The meeting took place in the New Conference Center Kirchberg with the support of the Fonds National de la Recherche (FNR, Luxembourg). This event gathered 48 renowned international speakers and more than 350 participants and 18 trade exhibitors.

Assessment of an RNA interference screen-derived mitotic and ceramide pathway metagene as a predictor of response to neoadjuvant paclitaxel for primary triple-negative breast cancer: a retrospective analysis of five clinical trials

Addition of taxanes to preoperative chemotherapy in breast cancer increases the proportion of patients who have a pathological complete response (pCR). However, a substantial proportion of patients do not respond, and the prognosis is particularly poor for patients with oestrogen-receptor (ER)/progesterone-receptor (PR)/human epidermal growth factor receptor 2 (HER2; ERBB2)-negative (triple-negative) disease who do not achieve a pCR. Reliable identification of such patients is the first step in determining who might benefit from alternative treatment regimens in clinical trials. We previously identified genes involved in mitosis or ceramide metabolism that influenced sensitivity to paclitaxel, with an RNA interference (RNAi) screen in three cancer cell lines, including a triple-negative breast-cancer cell line. Here, we assess these genes as a predictor of pCR to paclitaxel combination chemotherapy in triple-negative breast cancer. We derived a paclitaxel response metagene based on mitotic and ceramide genes identified by functional genomics studies. We used area under the curve (AUC) analysis and multivariate logistic regression to retrospectively assess the metagene in six cohorts of patients with triple-negative breast cancer treated with neoadjuvant chemotherapy; two cohorts treated with paclitaxel (n=27, 30) and four treated without paclitaxel (n=88, 28, 48, 39). The metagene was associated with pCR in paclitaxel-treated cohorts (AUC 0.79 [95% CI 0.53-0.93], 0.72 [0.48-0.90]) but not in non-paclitaxel treated cohorts (0.53 [0.31-0.77], 0.59 [0.22-0.82], 0.53 [0.36-0.71], 0.64 [0.43-0.81]). In multivariate logistic regression, the metagene was associated with pCR (OR 19.92, 2.62-151.57; p=0.0039) with paclitaxel-containing chemotherapy. The paclitaxel response metagene shows promise as a paclitaxel-specific predictor of pCR in patients with triple-negative breast cancer. The metagene is suitable for development into a reverse transcription-PCR assay, for which clinically relevant thresholds could be established in randomised clinical trials. These results highlight the potential for functional genomics to accelerate development of drug-specific predictive biomarkers without the need for training clinical trial cohorts. UK Medical Research Council; Cancer Research UK; the National Institute for Health Research (UK); the Danish Council for Independent Research-Medical Sciences (FSS); Breast Cancer Research Foundation (New York); Fondation Luxembourgeoise contre le Cancer; the Fonds National de la Recherche Scientifique; Brussels Region (IRSIB-IP, Life Sciences 2007) and Walloon Region (Biowin-Keymarker); Sally Pearson Breast Cancer Fund; and the European Commission.

The subchondral bone: a new frontier in articular cartilage repair

In the past decades, considerable advances have been made to treat defects of the articular cartilage. Yet the problem of lesions that extend deep in the underlying subchondral bone has not received adequate attention. Subchondral bone damage is mainly associated with four individual etiologies, including traumatic osteochondral fracture, osteochondritis dissecans, osteonecrosis, and osteoarthritis. It is largely because of these very different origins of osteochondral defects that their treatment remains complicated. In Mondorf-les-Bains, Luxembourg, from 8 to 10 October 2009, a group of researchers and physicians at the forefront of their field met to review and summarize the current state and potential future research directions of the subchondral bone in articular cartilage repair. This conference ‘‘The subchondral bone in articular cartilage repair’’ was co-chaired by the author and by Dr. Romain Seil (Luxembourg). The Deutsche Forschungsgemeinschaft (DFG) of Germany and the Fonds National de la Recherche (FNR) of Luxembourg jointly supported this endeavor. In this issue of Knee Surgery, Sports Traumatology, and Arthroscopy, the results of that meeting are presented as four individual review articles. They reflect not only a large body of current research advancement but also aim at identifying issues that still need to be answered. An initial article ‘‘Basic science of the subchondral bone’’ [2] reviews the anatomy, morphology, physiology, and pathology of the subchondral bone in the context of articular cartilage repair and in the light of the four different diseases. Evidence was shown that collagen fibrils cross the tidemark—the line that separates the calcified cartilage from the noncalcified cartilage—resulting in a strong link between these two zones. This is of clinical relevance—emphasizing the need for a meticulous removal of all calcified articular cartilage when a cartilage defect is debrided prior to a marrow-stimulating technique or autologous chondrocytes transplantation to allow for a good attachment of the repair tissue to the subchondral bone plate. The second article provides insights into the epidemiology and imaging of the subchondral bone [3]. Osteochondral defects, account for about 5% of all articular cartilage lesions, are predominantly located on the medial femoral condyle and patella and frequently associated with lesions of the menisci or the anterior cruciate ligament. Magnetic resonance imaging is the key procedure to evaluate articular cartilage and subchondral bone. However, when choosing an imaging technique to investigate osteochondral defects, the nature of the disease that causes the subchondral bone lesion has to be taken into account. For example, bone scintigraphy is valuable for early diagnosis of spontaneous osteonecrosis about the knee, while CT arthrography is more useful to evaluate the stability of the osteochondral fragment in osteochondritis dissecans. Another article is devoted to disease-specific clinical problems associated with the subchondral bone: osteochondral fracture, osteochondritis dissecans, osteonecrosis, and osteoarthritis [4]. If left untreated, all of them have the potential to progress from a prearthrotic deformity to joint destruction. Size and location of the subchondral lesion, stage of the disease, age of the patient, and congruency of the joint strongly influence the result of the surgical therapy. H. Madry (&) Institute for Experimental Orthopaedics and Department of Orthopaedic Surgery, Saarland University Medical Center, Saarland University, Homburg, Germany e-mail: hmad@hotmail.com

3 ANALYSIS OF HOX EXPRESSION IN BOVINE OOCYTES AND EARLY EMBRYOS

HOX genes encode transcription factors known to play a major role in patterning the main body axis of vertebrate embryos from the gastrulation stage onward. A few studies have provided evidence that some HOX genes might be expressed before implantation in mammalian embryos. Translation of maternally inherited transcripts is regulated by modifications of the poly(A) tail length until embryonic genome activation (EGA), occurring during the 4th cell cycle in the bovine. The objective of this work was to establish the expression pattern of various HOX genes and to study the polyadenylation of their transcripts during oocyte maturation and early embryonic development. Pools of 20 bovine oocytes before and after in vitro maturation and 20 in vitro-produced embryos at different stages of development up to the blastocyst stage were collected. Three to 12 pools were used for each stage. RNA was extracted and reverse transcribed (RT) using random hexamers. Quantitative real-time PCR (qPCR) was performed to establish expression profiles of 4 HOX genes: HOXD1, HOXA3, HOXB9, and HOXC9. Two distinct patterns of expression were observed. First, relative amounts of HOXD1, HOXA3, and HOXC9 were lower in morulae and blastocysts than in oocytes. On the other hand, relative expression of HOXB9 increased between the 5 to 8 cell stage and the morula stage (Mann-Whitney, P &lt; 0.05). Those expression patterns were not modified when embryos were cultured in presence of α-amanitin, a RNA polymerase II inhibitor, indicating the maternal origin of the transcripts until EGA. Total amount of mRNAs, estimated by RT-qPCR with random hexamers, was stable for all studied genes during oocyte maturation. The relative amount of polyadenylated GAPDH mRNAs, estimated by RT-qPCR with poly(dT), decreased greatly in mature oocytes compared with immature oocytes indicating massive deadenylation of those transcripts. The relative amount of polyadenylated HOXC9 transcripts decreased slightly but significantly during oocyte maturation (Mann-Whitney, P &lt; 0.05).The relative amount of polyadenylatedm RNAs corresponding to HOXD1, HOXA3, and HOXB9 was stable during oocyte maturation. This indicates that those transcripts escape the default deadenylation pathways followed by housekeeping genes. This experiment has been repeated 3 to 4 times. In conclusion, we confirmed the presence of HOXD1, HOXA3, HOXB9, and HOXC9 transcripts in bovine oocytes and early-stage embryos. Their role during oocyte maturation and the first stages of embryonic development will be investigated through loss of function studies. This work is funded by the Fonds National de la Recherche Scientifique (Belgium).

Foresight for science and technology priority setting in a small country: the case of Luxembourg

This paper sets out to review the conduct and immediate impacts of a two-year national technology foresight exercise (2006–2007) conducted in one of Europe’s smallest and wealthiest countries, Luxembourg. The country’s small size brings into sharp view many of the underlying tensions present in those foresight exercises that explicitly attempt to set national priorities. These tensions include the ability (or otherwise) to underpin the foresight process with sufficient and appropriate ‘objectivised’ knowledge (including national statistics, international benchmarking data, and future-oriented ‘intelligence’), the organisation of dialogic spaces that are not solely ‘hijacked’ by special interests and setting the ‘granularity’ of emergent priorities at a level that makes them ‘operationalisable’ in informing research and development funding programmes. The exercise was organised by the FNR (Fonds national de la recherche or National Research Fund), the only research council in Luxembourg. The paper shows that the position of the FNR in the research landscape had both benefits and drawbacks during the conduct of the exercise as well as for the follow-up implementation. The paper builds upon an earlier paper which described the first phase of the FNR Foresight exercise. The current paper extends this analysis to the second and third phases of the exercise, as well as to the immediate implementation phase after the publication of its results. By doing so, it covers the evolution of the foresight exercise over its two-year life span, highlighting the different meanings given to the exercise by different stakeholder groups as the process unfolded and interim results were made known. The paper draws lessons not only for other small countries and regions hoping to use foresight, but also highlights principles for using foresight for priority-setting more generally.

Power-Shaping Control of an Exothermic Continuous Stirred Tank Reactor (CSTR)* *The work of A. Favache is funded by a grant of aspirant of Fonds National de la Recherche Scientifique (Belgium).

Abstract Abstract The exothermic continuous stirred tank reactor (CSTR) is a classical yet complex case study of nonlinear dynamical systems. Power-shaping control is a recent approach for the control of nonlinear systems based on the physics of the dynamical system. In this paper we present a general methodology to apply the power-shaping control approach to the exothermic CSTR study case. It results in a global Lyapunov function for the exothermic CSTR. This Lyapunov function is then reshaped by the means of a controller in order to stabilize the process at a desired temperature. Some considerations on the local and global convergence to the desired state are presented.

CD8 + T-Cell Subpopulations in Human Abdominal Aortic Aneurysm Lesion

To the Editor: We read with interest the article by Duftner et al1 reporting the prevalence of peripheral interferon-γ (IFN-γ)–producing CD4+CD28− and CD8+CD28− T cells in patients with small abdominal aortic aneurysm (AAA). Along with the recent description that Th1-type immune responses predominate in human end-stage AAA lesion,2,3 their observation further supports preference toward polarized type 1 T-cell responses in aneurysm disease. The potential involvement of Th1 cells in the pathogenesis of the disorder is also suggested by the convincing demonstration that absence of CD4+ T cells or targeted deletion of IFN-γ prevents the induction of experimental AAA in a calcium chloride–induced mouse model,4 AAA formation being reconstituted by administration of IFN-γ into CD4−/− mice or infusion of competent splenocytes from wild-type mice into IFN-γ−/− mice. In their study, Duftner et al further established that both circulating CD4+CD28− and CD8+CD28− T cells are highly differentiated cells that display extensive CD45RO to CD45RA reversion and produce large amounts of IFN-γ and perforin. Surprisingly, low percentages of CD8+CD28− T cells were identified in AAA tissue sections using immunohistochemistry compared with flow cytometric analysis of peripheral blood mononuclear cells. In a series of our …

Un bouillon de culture pour les sciences de l'éducation? Le Congrès international d'éducation morale (1908–1934)

On aura donc à résoudre, par divers moyens, cette difficulté centrale qui consiste à parler d’une discipline qui n’existe pas encore, mais qui va exister, et pour laquelle il faut bien qu’il y ait, dans les temps de sa genèse, des éléments qui soient en train d’en préparer le surgissement. (Pierre Favre, Naissances de la science politique en France, 1870–1914 (Paris, 1989), p. 14) La recherche est soutenue par le Fonds national suisse de la recherche scientifique (1214-065300.01), dirigée par Rita Hofstetter et Bernard Schneuwly, avec la collaboration de Lucien Criblez, Valérie Lussi, Martina Späni et moi-même, soit l’Équipe de Recherche en Histoire des Sciences de l'éducation de l’Université de Genève (ERHISE). Je tiens à remercier les commentaires critiques que les membres de l’équipe ont apportés aux diverses versions du présent article. This article deals with the International moral education congress during its life period, 1908–1934. More specifically it examines the possible relationship between this Congress and the development of the educational science(s) as a disciplinary field. The study is mostly based on the proceedings and publications of the International moral education congresses. Inspired by historians/sociologists of sciences stressing the importance of the international dimension of scientific recognition, the analysis investigates the possible emergence of the educational science(s) in the context of the Congress. Four parts constitute this paper. A short introduction stresses first the problematic link between a congress devoted to educational matters, without exclusive scientific ambitions, and the emergence of a discipline of education, during the first decade of the twentieth century. The article then focuses on the aims and ideals of the instigators of the international moral. It points out the ambivalence between the internationality of the Congress, and the internationalism expressed by the activists of the movement, very close to the agenda of the international reconciliation, before and after the First World War. The second part goes into the proceedings of the six international congresses of the movement and gives a view of the socio-professional profiles of the congress participants. It deals with the dynamic tensions between on the one hand the scientific or academic dimensions of the movement (participants, status of the debates) and on the other the field of practice or administration in the education field. The third part concerns the content of the discussions held in the Congress, the general evolution of working themes and in particular the place occupied by university researchers or professors in education. Piaget declared as evidence in 1965 that the International moral education congress had been a vector of pedagogy as a scientific discipline. This article proposes to consider this Congress as a plausible but little productive culture medium of this emerging disciplinary field of education.

Radiocarbon Dates from the Oxford Ams System: Archaeometry Datelist 31

Radiocarbon Dates from the Oxford Ams System: <i>Archaeometry</i> Datelist 31

Vesicular Nanostructures Prepared by Reactive Melt Blending We are grateful to the Services Fédéraux des Affaires Scientifiques, Techniques et Culturelles for general support in the frame of “PAI 4-11: Supramolecular Chemistry and Supramolecular Catalysis”. We are grateful to Sunil K. Varshney (Polymer Source Inc., Canada) for training and advice in anionic polymerization and to Mrs. Martine Dejeneffe for ultramicrotomy at

Vesikel mit Schichtmorphologie: Beim Schmelzmischen von Polyamid 12 (PA12) mit symmetrischem Polystyrol-b-Polyisopren-Copolymer (PS-b-PIP) mit reaktiver Anhydrid-Endgruppe bilden sich Nanovesikel mit einer Dreischicht-Substruktur (siehe elektronenmikroskopische Aufnahme). Ist das Blockcopolymer unreaktiv, beobachtet man lediglich eine Separierung der Makrophasen.

Vesicular Nanostructures Prepared by Reactive Melt Blending We are grateful to the Services Fédéraux des Affaires Scientifiques, Techniques et Culturelles for general support in the frame of “PAI 4-11: Supramolecular Chemistry and Supramolecular Catalysis”. We are grateful to Sunil K. Varshney (Polymer Source Inc., Canada) for training and advice in anionic polymerization and to Mrs. Martine Dejeneffe for ultramicrotomy at

Vesicular Nanostructures Prepared by Reactive Melt Blending We are grateful to the Services Fédéraux des Affaires Scientifiques, Techniques et Culturelles for general support in the frame of “PAI 4-11: Supramolecular Chemistry and Supramolecular Catalysis”. We are grateful to Sunil K. Varshney (Polymer Source Inc., Canada) for training and advice in anionic polymerization and to Mrs. Martine Dejeneffe for ultramicrotomy at

Special issue on Econophysics

What is “Econophysics"? Who is an “econophysicist"? The coining of a new scientific term, composed of the names of two fields, traditionally considered to be far from each other, brings new dreams to investigators, by mere virtue of a new ensemble of viewpoints. The term “econophysics" has revealed a kinship between the fields of physics and economics, which was not obvious before. The first officially recognized conference by a professional society on “econophysics", Applications of Physics to Financial Analysis (APFA, soon to become APFA1) was held in Dublin in 1999. Since then APFA and its companion meetings have begun to reveal new branches of research from the established pathways explored in applied statistical physics and thus economics (in particular finance). The analysis of fluctuations in financial data by new or modified techniques has led to new insights. Such analysis involves physicists looking for correlation between entities in financial matter in much the same way as they have done for physical systems in their laboratories. This approach leads to useful new methods and results in different outputs. The studies of phase transitions and non-equilibrium effects, including self-organisation have progressed the understanding of many physical phenomena. So why not use the same methodology in a field which is thought to be governed by sociology, psychology, politics and other so called softer science? The observations of deterministic chaos, scaling, in financial time series (tools such as recurrence, plots exploiting symmetries in pricing theory or the use of the wavelet or path integral or renormalisation group method) will still give some work ahead even though all these tools have a basic origin or are rather standard tools nowadays. Characterization of data and theory talks broke new ground in pursuit of e.g. useful strategies or political consequences. One continues to ask, how is it that fluctuations or other agents in a system conspire to give surprising anomalous properties? By broadening discussion to the category of econophysics topics, as covered in APFA2 (held in Liege, Belgium on July 13-15, 2000), we have gained new paradigms to study this question. Several reports to APFA2 are not included in the following to avoid duplicating reports in this proceedings. Very warm and profound acknowledgments are in order here. APFA2 was made possible mainly by the European Physical Society (EPS), the Fond National de la Recherche Scientifique (FNRS, Brussels), the Fonds voor Wetenschappelijk Onderzoek-Vlaanderen (FWO), and the University of Liege.