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Vulvar varicose veins (VVs) are seen in 4% of women, most of them secondary to pregnancy and usually regressing spontaneously. The treatment of choice during pregnancy is conservative and symptomatic. Management of vulvar varicosities in non-pregnant women consists of various techniques, including phlebectomy, endovascular embolization or surgical ligation of contributing veins, sclerotherapy and, recently, conservative treatment with the venoactive agent MPFF (micronized purified flavonoid fraction). We report an unusual case of a large hematoma of the right labium majus following bilateral vulvar phlebectomy and embolization of the left ovarian vein. The patient was a non-pregnant woman, who underwent incision and drainage of this rare complication. At follow-up almost a year after this procedure the patient reported comfort and cosmetic satisfaction regarding her vulvar symptoms. A multidisciplinary team approach to vulvar varicosities is important, with the involvement of gynecologists, angiologists, interventional radiologists and vascular surgeons., Highlights • Vulvar varicose veins remains a uncommon disease with various treatments available, surgical as well as conservative • Serious complications after treatment of vulvar varicose veins have never been reported in the literature until today • We report a case of a large hematoma of the right labium majus following vulvar phlebectomy and ovarian vein embolization • The hematoma was successfully treated with surgical evacuation and the patient reported comfort and cosmetic satisfaction • Multidisciplinary team approach is essential in order to provide an optimal choice of treatment for vulvar varicose veins

Background: Pregnant women represent a vulnerable population at higher risk of complications of infectious diseases. Data regarding the consequences of the emerging pandemic severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) during pregnancy are scarce. Recent evidence suggests that pregnant women might be at higher risk of severe disease, while exposed fetuses and newborns could suffer from preterm birth, growth restriction and neonatal infections. Methods: We developed an international web registry to allow structured data collection. Pregnant women at any stage during gestation tested for SARS-CoV-2 infection were enrolled. Maternal, obstetrical and neonatal outcomes were recorded. Findings: 1033 pregnant women tested for SARS-CoV-2 were included, among which 926 tested positive and 107 tested negative. Positive pregnant women were at higher risk of severe maternal outcomes compared to negative women [aRR 5.6, 95% CI 1.4-22.7]. Risk factors for severe maternal outcomes among positive women were pulmonary comorbidities [aOR 4.3, 95% CI 1.9-9.5], hypertensive disorders [aOR 2.7, 95% CI 1.0-7.0] and diabetes [aOR2.2, 95% CI 1.1-4.5]. No difference in term of obstetrical and neonatal outcomes were observed between positive and negative women. Positive pregnant women with severe maternal outcomes were at higher risk of cesarean sections [70.7% (n=53/75)], preterm deliveries [62.7% (n= 32/51)] and newborns requiring admission to the neonatal intensive care unit [41.3% (n=31/75)]. A positive neonatal SARS-CoV-2 test was observed in 2.9% (n=11/384) of newborns with an available test at birth. Interpretation: Pregnant women, particularly those with associated comorbidities, seem to be at higher risk of severe complications of SARS-CoV-2 infection. Preliminary data regarding obstetrical and neonatal outcomes among women with a mild disease are reassuring. Funding Statement: None. Declaration of Interests: The authors declare that we have no conflicts of interest. Ethics Approval Statement: The study was approved by both the Swiss Ethical Board (CER-VD-2020-00548) and the local ethics boards at each participating center.

Background: Recent evidence suggests that pregnant women might be at higher risk of severe disease associated with the emerging pandemic severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), while exposed fetuses/newborns could suffer from preterm birth, growth restriction and neonatal infections. The magnitude of this increased risk and specific risk factors for severity remains unclear. Methods: We performed a case control study comparing pregnant women with severe coronavirus disease 19 (case) to pregnant women with a milder form (controls) enrolled in COVI-Preg international registry cohort between from March 24 to July 26, 2020. Risk factors for severity, obstetrical, fetal and neonatal outcomes were assessed. Findings: A total of 926 pregnant women with a positive test for SARS-CoV-2 were included, among which 92 (9.9%) presented a severe COVID-19 disease. Risk factors for severe maternal outcomes were pulmonary comorbidities [aOR 4.3, 95% CI 1.9-9.5], hypertensive disorders [aOR 2.7, 95% CI 1.0-7.0] and diabetes [aOR2.2, 95% CI 1.1-4.5]. Pregnant women with severe maternal outcomes were at higher risk of cesarean sections [70.7% (n=53/75)], preterm deliveries [62.7% (n= 32/51)] and newborns requiring admission to the neonatal intensive care unit [41.3% (n=31/75)]. Interpretation: Pregnant women, particularly those with associated comorbidities, seem to be at higher risk of severe complications of SARS-CoV-2 infection. Obstetrical and neonatal outcomes appear to be influenced by the severity of maternal disease; complications include cesarean sections, prematurity and neonatal admission to the intensive care unit. Funding Statement: None. Declaration of Interests: The authors declare that they have no conflicts of interest. Ethics Approval Statement: The study was approved by both the Swiss Ethical Board (CER-VD- 2020-00548) and the local ethics boards at each participating center.

STUDY QUESTION How should surgery for endometriosis be performed? SUMMARY ANSWER This document provides recommendations covering technical aspects of different methods of surgery for deep endometriosis in women of reproductive age. WHAT IS KNOWN ALREADY Endometriosis is highly prevalent and often associated with severe symptoms. Yet compared to equally prevalent conditions, it is poorly understood and a challenge to manage. Previously published guidelines have provided recommendations for (surgical) treatment of deep endometriosis, based on the best available evidence, but without technical information and details on how to best perform such treatment in order to be effective and safe. STUDY DESIGN, SIZE, DURATION A working group of the European Society for Gynaecological Endoscopy (ESGE), ESHRE and the World Endometriosis Society (WES) collaborated on writing recommendations on the practical aspects of surgery for treatment of deep endometriosis. PARTICIPANTS/MATERIALS, SETTING, METHODS This document focused on surgery for deep endometriosis and is complementary to a previous document in this series focusing on endometrioma surgery. MAIN RESULTS AND THE ROLE OF CHANCE The document presents general recommendations for surgery for deep endometriosis, starting from preoperative assessments and first steps of surgery. Different approaches for surgical treatment are discussed and are respective of location and extent of disease; uterosacral ligaments and rectovaginal septum with or without involvement of the rectum, urinary tract or extrapelvic endometriosis. In addition, recommendations are provided on the treatment of frozen pelvis and on hysterectomy as a treatment for deep endometriosis. LIMITATIONS, REASONS FOR CAUTION Owing to the limited evidence available, recommendations are mostly based on clinical expertise. Where available, references of relevant studies were added. WIDER IMPLICATIONS OF THE FINDINGS These recommendations complement previous guidelines on management of endometriosis and the recommendations for surgical treatment of ovarian endometrioma. STUDY FUNDING/COMPETING INTEREST(S) The meetings of the working group were funded by ESGE, ESHRE and WES. Dr Roman reports personal fees from ETHICON, PLASMASURGICAL, OLYMPUS and NORDIC PHARMA, outside the submitted work; Dr Becker reports grants from Bayer AG, Volition Rx, MDNA Life Sciences and Roche Diagnostics Inc. and other relationships or activities from AbbVie Inc., and Myriad Inc, during the conduct of the study; Dr Tomassetti reports non-financial support from ESHRE, during the conduct of the study; and non-financial support and other were from Lumenis, Gedeon-Richter, Ferring Pharmaceuticals and Merck SA, outside the submitted work. The other authors had nothing to disclose. TRIAL REGISTRATION NUMBER na

The endometrium is a complex multicellular tissue lining the uterus that plays a crucial role in human reproduction. This multi-layered organ is composed of a functionalis layer, adjacent to the uterine lumen and a basalis layer, attached to the myometrium. These layers comprise several cell types including luminal and glandular epithelial cells, endometrial stromal cells, immune cells and vascular cells forming the spiral arterioles (Kobayashi and Behringer, 2003; Roy and Matzuk, 2011; Hart, 2016). This unique tissue undergoes remarkable dynamic remodeling orchestrated by menstrual repair, proliferation and differentiation processes, which are carefully controlled by female sex hormones during the menstrual cycle. The remodeling requires a delicate interplay of cellular and molecular events that enable the endometrium to be receptive for embryo implantation. During the proliferative (or follicular) phase, both the endometrial glands and stroma of the functionalis layer proliferate in response to rising estrogen levels coming from the ovarian follicular. Consequently, the thickness of the endometrium increases. Then, the functionalis layers maturate during the secretory (or luteal) phase with the concomitant presence of secretory and ciliated cells in response to progesterone. In the absence of conception, this tissue sheds before regenerating for a subsequent menstrual cycle (Kobayashi and Behringer, 2003; Roy and Matzuk, 2011). Defects in endometrium remodeling and function can lead to the development of various types of disorders that affect considerable numbers of women. These includes infertility, pregnancy disorders, endometriosis and endometrial cancers (Hart, 2016). For instance, endometriosis a condition in which endometrial tissues proliferate outside the uterus leading to pelvic pain, excessive bleeding and infertility affects between 10 and 15% of all women of reproductive age (Giudice and Kao, 2004; Vercellini et al., 2013). Endometrial cancer, the most common malignancy of the female genital tract, affects ~3% of women, with the highest rates registered in North America and Europe (Lortet-Tieulent et al., 2017; Urick and Bell, 2019). Even if progress has been made in the knowledge of the endometrium structure and function, little is known regarding the molecular and the cellular mechanisms involved in this dynamic remodeling in both physiological and pathological conditions. The main obstacle of such studies is the lack of accurate models. Many insights have been provided thanks to studies using mouse models. However, these models do not accurately recapitulate the specificities of human endometrial development and function. As a matter of fact, endometrium decidualization in mice and humans are quite different. Decidualization of the endometrium occurs in rodents exclusively when there is an incoming embryo or in response to mechanical injury, whereas in humans the endometrium undergoes decidualization in a cyclic manner, regardless of the presence of an embryo (Gellersen et al., 2007; Peterse et al., 2018). Therefore, findings obtained with these animals often cannot be directly translated to humans. Immortalized or carcinoma-derived cell lines, such as Ishikawa cells (derived from epithelial endometrial cells) or St-T1b cell (derived from stromal endometrial cells) are also available for in vitro investigations. While these cells are easily cultured for long periods (Table 1), there are general limitations of using such cell lines including their genetic background, the potential changes acquired following transformation and during their establishment in culture (Mannelli et al., 2015). Alternatively, primary endometrial cells can be isolated, but these are difficult to maintain and expand in long term culture. Most of these cells lose their phenotype and hormone responsiveness in culture (Mannelli et al., 2015). Moreover, two dimensional (2D) cell cultures do not faithfully mimic in vivo three dimensional (3D) structure and function. These limitations prompted researchers to find novel strategies to model human endometrium. To that end, 3D-culture methods, namely organoids hold promise as models to better mimic in vivo human endometrium. Table 1 Comparative advantages and drawbacks of the current in vitro endometrial models.

Idiopathic pulmonary fibrosis (IPF) is a complex disease involving various cell types. Macrophages are essential in maintenance of physiological homeostasis, wound repair and fibrosis in the lung. Macrophages play a crucial role in repair and remodeling by altering their phenotype and secretory pattern in response to injury. The secretome of induced pluripotent stem cells (iPSC-cm) attenuates injury and fibrosis in bleomycin injured rat lungs. In the current study, we evaluate the effect of iPSC-cm on gene expression and phenotype of interstitial macrophage in bleomycin injured rat lungs in vivo. iPSC-cm was intratracheally instilled 7 days after bleomycin induced lung injury and assessed 7 days later and single cell isolation was performed. Macrophages were FACS sorted and microarray analysis was performed. We characterized changes in the rat lung interstitial macrophages using transcriptional profiling. iPSC-cm reduced the total collagen content of the lung and reduced different macrophage populations. Gene set enrichment analysis revealed involvement of three essential pathways (a) immune modulation, (b) branching morphogenesis and (c) canonical Wnt signaling. This study demonstrates that iPSC-cm reduces fibrosis in bleomycin injured rat lung by partially altering the macrophages and regulating their gene expression. Electronic supplementary material The online version of this article (10.1007/s12015-017-9790-9) contains supplementary material, which is available to authorized users.

Endometriosis is a curious pathology that has been the source of many international publications.

The authors report a case of pseudomyxoma peritonei with gelatinous peritoneum in a 47-year-old-woman. The main symptom for discovery was a chronic pelvic abdominal pain. This disease is particularly rare. The gelatinous substance is often associated with a malignant ovarian tumor or appendicitis perforated. Currently, on the whole, an exploratory laparoscopy allows diagnosis, biopsies, and appendectomy. The treatment is essentially surgical. The prognosis depends on grade (1/3) and response to chemotherapy. This case was presented to the tumor board.

Specialist training in reproductive medicine within Europe continues to evolve. Recent revisions, updates, and initiatives have helped to refine the core educational needs for the specialist trainee.

International audience; New SRAM bit cell architectures have been proposed recently as solutions to the limitations of the six-transistor (6T) SRAM bit cell in term of minimum supply voltage, VDDMIN. There is no demonstrated bit cell as superior under ultra-low supply voltage like the 6T bit cell at nominal voltage. Main limitations concern first the ratio between the read current and the standby current at the lowest operating voltage, second the bit cell robustness to perturbations and third the data sensing sensitivity, among other but minor limitations. The paper presents two proposals of ten-transistor (10T) Ultra-Low-Voltage bit cell for 0.3 V operation and processed in 28 nm LP CMOS bulk. Simulation results are compared to experimental results to demonstrate a satisfying operation at Ultra-Low supply voltage.

Down syndrome (DS, trisomy 21), is the most common viable chromosomal disorder, with an incidence of 1 in 800 live births. Its phenotypic characteristics include intellectual impairment and several other developmental abnormalities, for the majority of which the pathogenetic mechanisms remain unknown. In this “Data in Brief” paper, we sum up the whole genome analysis by mRNA sequencing of normal and DS induced pluripotent stem cells that was recently published by Hibaoui et al. in EMBO molecular medicine.

Though the number of Vesicouterine Fistulas is steadily increasing in association with the increasing number of cesarean sections, clinical presentation can be subtle and not easily identified. In this case study, we will review the presenting symptoms of this complication and suggest an updated management protocol.

STUDY QUESTION: What progress has been made in fertility preservation (FP) over the last decade?SUMMARY ANSWER: FP techniques have been widely adopted over the last decade and therefore the establishment of international registries on their short- and long-term outcomes is strongly recommended.WHAT IS KNOWN ALREADY: FP is a fundamental issue for both males and females whose future fertility may be compromised. Reproductive capacity may be seriously affected by age, different medical conditions and also by treatments, especially those with gonadal toxicity. There is general consensus on the need to provide counselling about currently available FP options to all individuals wishing to preserve their fertility.STUDY DESIGN, SIZE, DURATION: An international meeting with representatives from expert scientific societies involved in FP was held in Barcelona, Spain, in June 2015.PARTICIPANTS/MATERIALS, SETTING, METHODS: Twenty international FP experts belonging to the American Society of Reproductive Medicine, ESHRE and the International Society of Fertility Preservation reviewed the literature up to June 2015 to be discussed at the meeting, and approved the final manuscript. At the time this manuscript was being written, new evidence considered relevant for the debated topics was published, and was consequently included.MAIN RESULTS AND THE ROLE OF CHANCE: Several oncological and non-oncological diseases may affect current or future fertility, either caused by the disease itself or the gonadotoxic treatment, and need an adequate FP approach. Women wishing to postpone maternity and transgender individuals before starting hormone therapy or undergoing surgery to remove/alter their reproductive organs should also be counselled accordingly. Embryo and oocyte cryopreservation are first-line FP methods in postpubertal women. Metaphase II oocyte cryopreservation (vitrification) is the preferred option. Cumulative evidence of restoration of ovarian function and spontaneous pregnancies after ART following orthotopic transplantation of cryopreserved ovarian tissue supports its future consideration as an open clinical application. Semen cryopreservation is the only established method for FP in men. Testicular tissue cryopreservation should be recommended in pre-pubertal boys even though fertility restoration strategies by autotransplantation of cryopreserved testicular tissue have not yet been tested for safe clinical use in humans. The establishment of international registries on the short- and long-term outcomes of FP techniques is strongly recommended.LIMITATIONS, REASONS FOR CAUTION: Given the lack of studies in large cohorts or with a randomized design, the level of evidence for most of the evidence reviewed was 3 or below.WIDER IMPLICATIONS OF THE FINDINGS: Further high quality studies are needed to study the long-term outcomes of FP techniques.STUDY FUNDING/COMPETING INTEREST(S): None.TRIAL REGISTRATION NUMBER: N/A.

We report the case of a 45-year-old woman who palpated a voluminous painless lump in the superior outer quadrant of her left breast. Her past medical history revealed an acute myeloid leukemia treated and considered in remission 1 month prior to this discovery. Imaging work-up by mammogram, US and MRI showed multiples masses suspect of malignancy in both breasts. US-guided needle biopsy was performed in the palpable mass and in one of the multiple lesions located in the right breast. Histologic findings were compatible with a granulocytic sarcoma in both breasts which was considered as a relapse of the acute myeloid leukemia treated a few months earlier.

Introduction Pulp repair is less likely to occur when dentin or pulpal tissue remains infected after caries excavation. Yet there are currently few options to kill residual bacteria without damaging resident cells. The current study has evaluated the effect of 3 blue light–activated chemicals on the viability of lactobacilli, odontoblast-like cells (MDPC-23), undifferentiated pulp cells (OD21), and human embryonic stem cells (hESC H1). Methods Bacteria were incubated for 15 minutes with curcumin, eosin Y, or rose bengal and then irradiated with blue light (240 seconds). Bacteria were labeled with LIVE/DEAD BacLight Bacterial Viability kit, and viability was assessed by fluorescence-activated cell sorting. Cytotoxicity assays were performed on MDPC-23 cells, OD21, and hESC H1 cells grown in 24-well plates and exposed to the same photosensitizer-light combination. After 24 hours, cellular response was measured by using the methyl-thiazol-diphenyl-tetrazolium assay. Results were statistically analyzed by using one-way analysis of variance and Tukey multiple comparison intervals (α = 0.05). Results Bacterial viability was significantly reduced after exposure to different combinations of light and photosensitizers; mitochondrial activity of cultured cells remained unaffected when exposed to the same conditions, suggesting a good therapeutic index in vitro. Conclusions Blue light–mediated disinfection is promising for the development of new treatment strategies designed to promote pulp repair after carious exposure.

The establishment of an alveolar epithelial cell (AEC) line derived from induced pluripotent stem cells (iPSC) represents a novel opportunity in lung disease modelling and tissue engineering. In the present study, we succeeded in generating type II AECs from iPSCs. To ensure homogeneous and long term propagation of these cells, iPSC-derived type II AECs, were immortalized by human telomerase reverse transcriptase andpolycomb complex protein BMI-1 lentiviral vectors. The generated type II AEC line displayed morphological characteristics of type II AECs including growing as a cobble stone monolayer. Thus, electron microscopy analysis of this cell line confirmed the presence of lamellar body and microvilli. Also this cell line expressed type II AEC proteins such as cytokeratin, surfactant protein C and lysotracker DND 26 (marker for lamellar body). Furthermore, the type II AEC line exhibited functional properties of type II AECs including an increase of their transepithelial electrical resistance over time and an improved wound closure in presence of serum in the invitro wound healing assay. Upon TNF-α stimulation, these cells released IL-6 an IL-8 cytokines. Consistent with type II AEC phenotype, the cell line showed the ability to uptake and release of surfactant protein B and to differentiate towards type I AECs. Altogether, this study established immortalized type II AECs derived from iPSCs as a novel cellular model for lung disease modelling and tissue engineering.

Pulmonary fibrosis is a progressive lung disease, resulting in death of the patient. Macrophages play a critical role in tissue remodeling in this disease. Recently, we have shown that secretome of induced pluripotent stem cells (iPSC-cm) attenuates fibrosis in bleomycin injured rat lungs. We hypothesize that iPSC-cm exerts its antifibrotic effect by altering the lung macrophage transcription profile due to multiple factors present in the secretome. Adult male rats were instilled with bleomycin, and were treated by iPSC-cm or control media. Macrophages were sorted out and cDNA was synthesized. Differential genes expression was determined using Agilent microarrays. The protein composition of the iPSC-cm was examined using LC/MS2 shotgun proteomics. Both datasets were integrated using the STRING protein interaction database. Percentage of total macrophages increased in bleomycin injured lungs compared to normal control (26.2±1.4% vs 14.4±1.0%). iPS-cm treatment reduced total percentage of macrophages to 17.4±1.2%; moreover, the original macrophage phenotype was partially restored (M1: 18.0%±8.3% vs 32.3±2.0%; M2: 1.3%±0.7 vs. 5.7%±0.3%). Pathway analysis of the microarray data indicated that treatment with iPSC-cm specifically regulates genes involved in the organization of the basal membrane. Subsequent network analysis identified several factors in the secretome that could be responsible for the observed antifibrotic effect.iPSC-cm is rich in many antifibrotic molecules that plays critical role in changing macrophage phenotype thus helping in tissue remodeling and reducing fibrosis. This abstract has been presented previously at the European Respiratory Society9s Lung Science Conference in March 2016.

Minimally invasive surgery (MIS) can be considered as the greatest surgical innovation over the past thirty years. It revolutionized surgical practice with well-proven advantages over traditional open surgery: reduced surgical trauma and incision-related complications, such as surgical-site infections, postoperative pain and hernia, reduced hospital stay, and improved cosmetic outcome. Nonetheless, proficiency in MIS can be technically challenging as conventional laparoscopy is associated with several limitations as the two-dimensional (2D) monitor reduction in-depth perception, camera instability, limited range of motion and steep learning curves. The surgeon has a low force feedback which allows simple gestures, respect for tissues and more effective treatment of complications. Since 1980s several computer sciences and robotics projects have been set up to overcome the difficulties encountered with conventional laparoscopy, to augment the surgeon's skills, achieve accuracy and high precision during complex surgery and facilitate widespread of MIS. Surgical instruments are guided by haptic interfaces that replicate and filter hand movements. Robotically assisted technology offers advantages that include improved three- dimensional stereoscopic vision, wristed instruments that improve dexterity, and tremor canceling software that improves surgical precision.

We report a case of woman with a palpable lump on her left breast. On mammography a huge mass located between the inner and outer inferior breast quadrants of the left breast was found. The ultrasound examination realized later revealed a heterogeneous mass with smooth and lobulated borders. A MRI was also performed, showing an oval mass with heterogeneous areas of enhancement. Finally a core biopsy under sonographic guidance revealed a pseudo angiomatous stromal hyperplasia of the breast (PASH).

Neuronal disorders, like Huntington's disease (HD), are difficult to study, due to limited cell accessibility, late onset manifestations, and low availability of material. The establishment of an in vitro model that recapitulates features of the disease may help understanding the cellular and molecular events that trigger disease manifestations. Here, we describe the generation and characterization of a series of induced pluripotent stem (iPS) cells derived from patients with HD, including two rare homozygous genotypes and one heterozygous genotype. We used lentiviral technology to transfer key genes for inducing reprogramming. To confirm pluripotency and differentiation of iPS cells, we used PCR amplification and immunocytochemistry to measure the expression of marker genes in embryoid bodies and neurons. We also analyzed teratomas that formed in iPS cell-injected mice. We found that the length of the pathological CAG repeat did not increase during reprogramming, after long term growth in vitro, and after differentiation into neurons. In addition, we observed no differences between normal and mutant genotypes in reprogramming, growth rate, caspase activation or neuronal differentiation. However, we observed a significant increase in lysosomal activity in HD-iPS cells compared to control iPS cells, both during self-renewal and in iPS-derived neurons. In conclusion, we have established stable HD-iPS cell lines that can be used for investigating disease mechanisms that underlie HD. The CAG stability and lysosomal activity represent novel observations in HD-iPS cells. In the future, these cells may provide the basis for a powerful platform for drug screening and target identification in HD.

Stem cells guarantee tissue repair and regeneration throughout life. The decision between cell self-renewal and differentiation is influenced by a specialized microenvironment called the ‘stem cell niche’. In the tooth, stem cell niches are formed at specific anatomic locations of the dental pulp. The microenvironment of these niches regulates how dental pulp stem cell populations participate in tissue maintenance, repair, and regeneration. Signaling molecules such as Notch proteins are important regulators of stem cell function, with various capacities to induce proliferation or differentiation. Dental injuries often lead to odontoblast apoptosis, which triggers activation of dental pulp stem cells followed by their proliferation, migration, and differentiation into odontoblast-like cells, which elaborate a reparative dentin. Better knowledge of the regulation of dental pulp stem cells within their niches in pathological conditions will aid in the development of novel treatments for dental tissue repair and regeneration.

Down syndrome (DS) caused by a trisomy of chromosome 21 (HSA21), is the most common genetic developmental disorder, with an incidence of 1 in 800 live births. Its phenotypic characteristics include intellectual impairment, early onset of Alzheimer’s disease, congenital heart disease, hypotonia, muscle weakness and several other developmental abnormalities, for the majority of which the pathogenetic mechanisms remain unknown. Among the numerous protein coding genes of HSA21, dual-specificity tyrosine-(Y)-phosphorylation-regulated kinase 1A (DYRK1A) encodes a proline-directed serine/threonine and tyrosine kinase that plays pleiotropic roles in neurodevelopment in both physiological and pathological conditions. Numerous studies point to a crucial role of DYRK1A protein for brain defects in patients with DS. Thus, DYRK1A inhibition has shown benefits in several mouse models of DS, including improvement of cognitive behaviour. Lastly, a recent clinical trial has shown that epigallocatechine gallate (EGCG), a DYRK1A inhibitor, given to young patients with DS improved visual recognition memory, working memory performance and adaptive behaviour.

background: Both human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) bear a great potential in regenerative medicine. In addition to optimized clinical grade culture conditions, efficient clinical grade cryopreservation methods for these cells are needed. Obtaining good survival after thawing has been problematic. methods: We used a novel, chemically defined effective xeno-free cryopreservation system for cryostorage and banking of hESCs and iPSCs. The earlier established slow freezing protocols have, even after recent improvements, resulted in low viability and thawed cells had a high tendency to differentiate. The medium is a completely serum and animal substance free product containing dimethylsulfoxide, anhydrous dextrose and a polymer as cryoprotectants. The cells were directly frozen at 2708C, without a programmed freezer. results: The number of frozen colonies versus the number of surviving colonies differed significantly for both HS293 (x 2 ¼ 9.616 with one degree of freedom and two-tailed P ¼ 0.0019) and HS306 (x 2 ¼ 8.801 with one degree of freedom and two-tailed P ¼ 0.0030). After thawing, the cells had a high viability (90– 96%) without any impact on proliferation and differentiation, compared with the standard freezing procedure where viability was much lower (49%). The frozen –thawed hESCs and iPSCs had normal karyotype and maintained properties of pluripotent cells with corresponding morphological characteristics, and expressed pluripotency markers after 10 passages in culture. They formed teratomas containing tissue components of the three germ layers. conclusion: The defined freezing –thawing system described here offers an excellent simple option for banking of hESCs and iPSCs.

Embryonic stem (ES) cells are powerful tools to understand mechanisms of neuronal differentiation and to engineer neurons for in vitro studies and cell therapy. We developed a screening approach to identify small organic molecules driving neuronal differentiation of ES cells. For this purpose, we used a lentivector carrying a dual luciferase reporter system to engineer an ES cell line which allowed us to screen for small organic molecules enhancing neuronal differentiation. One of them, phenazopyridine, was further analysed in human ES cells. Phenazopyridine: (i) enhanced neuronal differentiation, (ii) increased cell survival, (iii) decreased the amount of non-neuronal and undifferentiated cells and (iv) synchronized the cellular differentiation state. Phenazopyridine allowed the development of a differentiation protocol compatible with the generation of clinical grade neural precursors, which were able differentiate into different neuronal subtypes, astrocytes and oligodendrocytes. In summary, we describe a powerful approach to identify small molecules directing stem cell differentiation. This led to the establishment of a new application for an old drug and the development of a novel clinical grade protocol for neuronal differentiation of ES cells.

BACKGROUND: Feeder cells are frequently used for the early-stage of derivation and culture of human embryonic stem cell (hESC) lines. METHODS: We established a conditionally immortalized human foreskin fibroblast line that secreted basic fibroblast growth factor (bFGF). These cells were used as feeder cells for hESC culture and induced pluripotent stem (iPS) cell derivation and expansion. This conditional immortalization was performed using lentiviral vector (LV) mediated transduction of Bmi-1 and human telomerase reverse transcriptase genes and the resulting cell line was further modified by LV-mediated transduction of a secreted form of bFGF gene product. Three different laboratories have tested whether this feeder cell line could support the maintenance of four different hESC lines. RESULTS: Immortalized fibroblasts secreting stable amounts of bFGF supported the growth of all hESC lines, which remained pluripotent and had a normal karyotype for at least 10 passages. Even at high passage (p56), these modified cells, when used as feeders, could support iPS derivation and propagation. Derived iPS cells expressed pluripotency markers, had hESC morphology and produced tissue components of the three germ layers when differentiated in vitro. CONCLUSION: These modified fibroblasts are useful as a genetically-defined feeder cell line for reproducible and cost-effective culture of both hESC and iPS cells.

Cell replacement therapy is a widely discussed novel concept of medical treatment. The increased knowledge in the stem cell field, particularly pluripotent stem cells, potentially provides powerful tools for this therapeutic concept. A large number of disease characterized by the loss of functional cells are potential candidates for cell replacement therapy and, in this regards, Parkinson's disease is of particular interest. It is one of the most prevalent neurodegenerative diseases caused by the loss of dopaminergic neurons in the Substantia nigra pars compacta. Pharmacological therapies are valuable but suffer from the progressive decline of efficacy as the disease progresses. Cell therapy application has emerged about two decades ago as a valid therapeutic alternative and recent advances in stem cell research suggest that pluripotent stem cell transplantation may be a promising approach to replace degenerated neurons in Parkinson's disease. Various sources of pluripotent stem cells (PSC) currently tested in animal models of Parkinson's disease have proven their efficacy in relieving symptoms and restoring damaged brain function. This review summarizes and discusses the important challenges that actually must be solved before the first studies of PSC transplantation can be undertaken into humans.

Human embryonic stem cells (hESC) not only hold great promise for the treatment of degenerative diseases but also provide a valuable tool for developmental studies. However, the clinical applications of hESC are at present limited by xeno-contamination during the in vitro derivation and propagation of these cells. In this review, we summarize the current methodologies for the derivation and the propagation of hESC in conditions that will eventually enable the generation of clinical-grade cells for future therapeutic applications.

Incontinence is a major public health concern in aging societies. It is caused by age-dependent spontaneous apoptosis of muscle cells in the urinary and fecal sphincters, and is aggravated in women due to birth trauma. Compared to other currently employed invasive surgical management techniques associated with morbidity and recurrence, replacement or regeneration of dysfunctional sphincter through stem cell therapy and tissue engineering techniques hold great promise. This review focuses on the pathophysiological analysis of urinary incontinence and the possible application of muscle-derived-stem cells, satellite cells, chondrocytes and adipose-derived-stem cells in restoring sphincter functions.

Background: Macrophages are involved in the pathogenesis and progression of pulmonary fibrosis (PF). Imbalance between M1 and M2 macrophages could play role in progression of PF. The secretome from human induced pluripotent stem cells (iPS) reduce bleomycin induced fibrosis in the rat lung. We investigate the effect of iPS conditioned media (iPS-cm) on macrophages. Methods: Human monocytes were polarized towards M1 and M2 phenotype, and treated with iPS-cm; migration and secretory properties were tested in vitro . Bleomycin injured rats were treated on d7 either with iPSC-cm or CCD1-cm; FACS analysis was performed on d14. Results: iPS-cm increased migration of macrophages in vitro . Levels of proinflammatory and profibrotic mediators were reduced after in vitro treatment with iPSC-cm. In vivo , percentage of total macrophages increased in bleomycin injured lungs compared to control. Moreover, M1 and M2 populations were both increased in bleomycin group. Interestingly, iPS-cm treatment reduced total percentage of macrophage; moreover, the original macrophage phenotype was partially restored. These effects couldn9t be detected using CCD1-cm. Conclusions: iPS-cm modulates migration and alters the macrophage phenotype by reducing pro inflammatory and pro fibrotic mediators in vitro and alters macrophage percentage and phenotype in vivo in a lung injury and fibrosis model. Targeting the macrophage phenotypical switch could be a possible therapy for PF.

Polycystic ovary syndrome (PCOS) is a frequent disorder, affecting approximately 5 to 10 % of infertile women. It can represent more than 80% of cases of infertility due to anovulation. The main goal of treatment is the induction of mono-ovulatory cycles. A pragmatic management of infertility in PCOS will allow most patients to conceive.Weight loss and clomiphene citrate (CC) are the first-line components of patients treatment before gonadotropins are used. However, during gonadotropin administration, there is a high risk of ovarian hyper-stimulation (OHSS) and multiple pregnancies. So, surgery with laparoscopic ovarian drilling (LOD) is often used before gonadotropins in order to obtain normal ovulatory cycles.

Objectives The aim of this mini review is to determine the relationship between endometriosis and urinary tract symptoms and to investigate the consequences of surgical treatment of mild to severe endometriosis, especially deep lesions, on the vesico-sphincteral function (lower urinary tract function). Materials and methods We performed a literature review by searching the MEDLINE database for articles published between 2000 and 2014, limiting the searches to the words: urinary tract, vesico-sphincteral, dysfunction, endometriosis, symptoms, and surgery. Results The incidence of vesico-sphincteral symptoms in endometriosis varies from 3.4 up to 15.4%. The frequency of such symptoms seems to be under estimated because of a lack of specific questionnaire including urinary symptoms. Urodynamic evaluation could help to detect unsuspected abnormalities. It seems that endometriosis surgery (particularly deep infiltrating lesions) is a purveyor of de novo urinary dysfunction, with an incidence varying from 6.8 up to 17.5%. Nerve sparing processes such as neuro-navigators or neuro-stimulators seem to be promising techniques to avoid postoperative urinary tract dysfunction. Conclusion A precise anamnesis and the use of specific validated questionnaires (IPSS and BFLUTS) improve the screening of vesico-sphincteral symptoms in case of endometriosis. No recommendation can be found in the literature about the place of urodynamic evaluation. Most publications lack of proof and therefore do not allow making recommendations about optimal treatment of endometriotic lesions to avoid urinary tract disorders.

Down syndrome (DS, trisomy 21), is the most common viable chromosomal disorder, with an incidence of 1 in 800 live births. Its phenotypic characteristics include intellectual impairment and several other developmental abnormalities, for the majority of which the pathogenetic mechanisms remain unknown. Several models have been used to investigate the mechanisms by which the extra copy of chromosome 21 leads to the DS phenotype. In the last five years, several laboratories have been successful in reprogramming patient cells carrying the trisomy 21 anomaly into induced pluripotent stem cells, i.e., T21-iPSCs. In this review, we summarize the different T21-iPSCs that have been generated with a particular interest in the technical procedures and the somatic cell types used for the reprogramming.

The transcription factor Pax6 plays an important role in the development of the central nervous system. To understand its mechanism of action, we transduced HeLa cells with a Pax6-expressing lentiviral vector. Upon transduction, HeLa cells markedly changed shape and formed neuritelike extensions. Pax6-transduced HeLa cells expressed high levels of neuronal alpha3 tubulin, demonstrating a partial transdifferentiation towards a neuronal phenotype. Neurons are postmitotic cells. Pax6-transduced HeLa cells became postmitotic through mechanisms involving up-regulation of p53 and cyclin-dependent kinase inhibitor p21. One of the most striking effects of Pax6 was observed by time-lapse videomicroscopy: cells started to dissociate from cell clusters and displayed intense migratory activity. Migration was accompanied by dynamic and reversible shape changes. Our results identified three elements of Pax6 action: (i) expression of neuron-specific genes; (ii) establishment of a postmitotic phenotype; and (iii) involvement in the regulation of cell shape and cell migration.

Introduction Idiopathic pulmonary fibrosis (IPF) is a progressive and irreversible fibrotic lung disease, resulting in respiratory insufficiency and reduced survival. Pulmonary fibrosis is a result of repeated alveolar epithelial microinjuries, followed by abnormal regeneration and repair processes in the lung. Recently, stem cells and their secretome have been investigated as a novel therapeutic approach in pulmonary fibrosis. We evaluated the potential of induced pluripotent stem cells (iPSC) conditioned media (iPSC-cm) to regenerate and repair the alveolar epithelium in vitro and improve bleomycin induced lung injury in vivo. Methods IPSC-cm was collected from cultured iPSC derived from human foreskin fibroblasts and its biological effects on alveolar epithelial wound repair was studied in an alveolar wound healing assay in vitro. Furthermore, iPSC-cm was intratracheally instilled 7 days after bleomycin induced injury in the rat lungs and histologically and biochemically assessed 7 days after instillation. Results iPSC-cm increased alveolar epithelial wound repair in vitro compared with medium control. Intratracheal instillation of iPSC-cm in bleomycin-injured lungs reduced the collagen content and improved lung fibrosis in the rat lung in vivo. Profibrotic TGFbeta1 and α-smooth muscle actin (α-sma) expression were markedly reduced in the iPSC-cm treated group compared with control. Antifibrotic hepatocyte growth factor (HGF) was detected in iPSC-cm in biologically relevant levels, and specific inhibition of HGF in iPSC-cm attenuated the antifibrotic effect of iPSC-cm, indicating a central role of HGF in iPSC-cm. Conclusion iPSC-cm increased alveolar epithelial wound repair in vitro and attenuated bleomycin induced fibrosis in vivo, partially due to the presence of HGF and may represent a promising novel, cell free therapeutic option against lung injury and fibrosis. Electronic supplementary material The online version of this article (doi:10.1186/scrt513) contains supplementary material, which is available to authorized users.

The BRCA1-associated RING domain protein BARD1 acts with BRCA1 in double-strand break repair and ubiquitination. BARD1 plays a role as mediator of apoptosis by binding to and stabilizing p53, and BARD1-repressed cells are resistant to apoptosis. We therefore investigated the mechanism by which BARD1 induces p53 stability and apoptosis. The apoptotic activity of p53 is regulated by phosphorylation. We demonstrate that BARD1 binds to unphosphorylated and serine-15 phosphorylated forms of p53 in several cell types and that the region required for binding comprises the region sufficient for apoptosis induction. In addition, BARD1 binds to Ku-70, the regulatory subunit of DNA-PK, suggesting that the mechanism of p53-induced apoptosis requires BARD1 for the phosphorylation of p53. Upregulation of BARD1 alone is sufficient for stabilization of p53 and phosphorylation on serine-15, as shown in nonmalignant epithelial cells and ovarian cancer cells, NuTu-19, which are defective in apoptosis induction and express aberrant splice variants of BARD1. Stabilization and phosphorylation of p53 in NuTu-19 cells, as well as apoptosis, can be induced by the exogenous expression of wild-type BARD1, suggesting that BARD1, by binding to the kinase and its substrate, catalyses p53 phosphorylation.

The BRCA1-binding RING-finger domain protein BARD1 may act conjointly with BRCA1 in DNA repair and in ubiquitination, but it may also induce apoptosis in a BRCA1-independent manner. In this study, we have investigated BARD1 expression during spermatogenesis. In contrast with BRCA1, which is expressed only in meiotic spermatocytes and early round spermatids, BARD1 is expressed during all stages of spermatogenesis. However, while spermatogonia expressed full-length BARD1 mRNA, later stages of spermatocyte precursors express predominantly a novel, shorter splice form BARD1beta. BARD1beta lacks the BRCA1-interacting RING finger but maintains its proapoptotic activity. Consistently, BRCA1 can counteract the proapoptotic activity of full-length BARD1 but not of BARD1beta. Several lines of evidence suggest that BARD1 is involved in proapoptotic signaling in testis: i) both BARD1 isoforms are mostly found in cells that stain positive for TUNEL, Bax, and activated caspase 3; ii) BARD1beta, capable of inducing apoptosis even in the presence of BRCA1, is specifically expressed in BRCA1-positive later stages of spermatogenesis; iii) antiapoptotic hormonal stimulation leads to BARD1 downregulation; and iv) BARD1 expression is associated with human pathologies causing sterility due to increased germ cell death. Our data suggest that full-length BARD1 might be involved in apoptotic control in spermatogonia and primary spermatocytes, while a switch to the BRCA1-independent BARD1beta might be necessary to induce apoptosis in BRCA1-expressing meiotic spermatocytes and early round spermatids.

Placental site trophoblastic tumor is rare. They represent a rare form of gestational trophoblastic disease. They occur mainly in women who have a history of miscarriage, termination of pregnancy, or even a normal or pathological ongoing pregnancy. The clinical course is unpredictable. This malignancy has different characteristics from other gestational trophoblastic tumors. Following a clinical case that we encountered and treated, we conducted a literary research and review, focusing primarily on prognostic factors and treatment.

Endometriosis is defined as the presence of endometrial-like tissue (glands or stroma) outside the uterus, which induces a chronic inflammatory reaction. Although endometriosis impairs fertility, it does not usually completely prevent conception. The question of evidence based-medicine guidelines in endometriosis-associated infertility is week in many situations. Therefore, we will highlight in this issue where the challenges are.

The scientific progresses achieved, during the twenty-first century, in molecular biology, in cytogenetics, in the use of stem cells in the treatment of various pathologies by reprograming them (1), in medical imaging and in surgical instrumentation allow an avant-gardist management in the fields of assisted reproductive medicine, in oncology, in gynecological surgery, and even in obstetrics. Therefore, couples can benefit from a specific and personalized management. This management is supervised by multidisciplinary teams respecting laws of medical ethics and those of bio-ethics. Should we still use the term of “sterility” when such a therapeutic armada is at our disposal? Born in the 70s, the laparoscopic surgery, now offers a better topographic approach of lesions, an almost perfect macroscopic depiction of colors. Hence, surgeons can spare more healthy tissues such as vessels and nerves, and thus adapt a more conservative procedure while restoring normal anatomy that is essential for preserving fertility. Endometriosis is one of the most obvious examples (2, 3). These advancements were achieved owing to new 3D imaging technologies and to the use of CO2 laser. The advent of new energies, i.e., RF, YAG laser, Argon laser, plasma energy, excimer, etc., opened new perspectives for surgical management of diseases. The miniaturization of instruments in hysteroscopy coupled with the development of hydrosonography allowed a better exploration of the uterine cavity, endometrial biopsies, and resection of intra-uterine lesions. Thus preventing invasive and radical treatments and reducing post-operative morbidity. However, the greatest advancements have been realized in cellular and molecular biology, in cytogenetics, and in proteomics. Indeed, the recent knowledge in the field of stem cells and their specific reprograming expand horizons to gene and cell therapy. A simple cell could be considered as a treatment by re-colonizing unhealthy tissues, thus preventing transplantation (1, 4, 5). The cryopreservation of ovarian tissue by vitrification before radiotherapy or chemotherapy in case of cancer gives a new hope to those couples touched by the disease. The in vitro culture of pathological cells, disease modeling, and collection of withdrawn tissues pre-operatively allows researchers to understand etiophysiopathogeny, and natural evolution of the disease as in endometriotic tissues (2). These procedures will avoid long, expensive, and ethically questionable studies on animal models, thus allowing pharmacotesting. Therefore, this will help defining new disease classification and propose therapeutic algorithms. Owing to progress in the field of infertility and assisted reproductive techniques (ART), i.e., intra cytoplasmic spermatozoid injection (ICSI) and vitrification of oocytes and embryos, increasing numbers of patients could be treated. In vitro endometrial culture leads to identification of reasons for treatment failure. Regarding pre-implantation genetic diagnosis (PGD), the progress in the knowledge of genetic mapping offers much hope to patients and couples suffering from family and hereditary diseases. A better knowledge of human papilloma viruses (HPV) and the conception of vaccination against those HPV contribute in the reduction of the prevalence of cervical cancer, since HPV is involved in the genesis of cervical dysplasia and thus in that of cervical cancer. A radical treatment of cervical dysplasia (surgical or laser CO2 conization) prevents their transformation toward an invasive cervical cancer. The use of CO2 laser coupled with colposcopy allowed “see-and-treat” management of cervical, vaginal, vulvar, and perineal lesions related to HPV in an ambulatory way with a minimal morbidity. Eradication of cervical cancer during this century is possible and conceivable, only if national programs of information of gynecologists, family practitioners, as well as a program of HPV vaccination are settled. In case of voiding dysfunction, stress urinary incontinence, overactive bladder, the innovations in the field of pressure sensors improved urodynamic measurements. A new clinical approach based on an itemized analysis of symptoms associated with progress of urodynamic tools allows a better therapeutic approach including both surgery and medical treatment. MRI, computerized mammography, 3D ultrasound, stereotactic, and ultrasound guided biopsies of breast lesions eased and fastened the diagnosis of breast cancer, with less morbidity. All these imaging techniques contributed to a better management of gynecological cancers and thus improved survival rates in these patients. Sentinel axillary lymph node biopsy with intra-operative frozen section allows to ovoid the systematically axillary lymph node resection. Protocols such as “ONCOTYPE,” based on cellular phenotyping, predict the need for chemotherapy. As an example, Her-2–Neu is used as a marker of poor prognosis and indicator for the use of HERCEPTIN neoadjuvant treatment. Radiotherapy becomes more focused on the tumor site, thus avoiding overdoses of irradiation and reducing side effects. Hormonal therapy is possible due to immunochemistry analysis of tumor-estrogen and -progesterone receptors, thus offering a longer period without recurrence. In case of mixed ovarian tumors, immunohistochemistry allows an accurate diagnosis. In the fields of contraception, a lot of progress has been achieved such as Levonorgestrel intra-uterine devices, progestogens of third and fourth generation allowing a decrease in ethinyl-estradiol dosage, progestogens implants, vaginal rings, and transdermal patches. So women have a very wide choice and contraception can be individually adapted. Gynecology is in constant progress allowing an ambulatory management with less morbidity thus reduced health costs, an earlier diagnosis with a better adapted and personalized treatment leading to higher survival rates. Obstetrics also knew considerable progresses. Since the twentieth century, prenatal diagnosis of congenital malformations and genetic diseases is made easier and earlier, a tribute of progresses in the fields of ultrasound, amniocentesis, and chorionic villi sampling (CVS), thus allowing an optimal management with less morbidity and less psychological disturbance in the respect of national legal frames. These great achievements could not be possible without progress in genetics and biology. Many markers are currently studied to screen earlier patients predisposed to develop preeclampsia during pregnancy or to predict premature rupture of membranes and labor, thus preventing wet lung syndrome. Progresses in neonatal resuscitation decreased perinatal morbidity and mortality. The detection of fetal DNA in the maternal blood with the hope of the complete genome fetal sequencing represents another breakthrough. Henceforth, in utero fetal surgery as a vesico-amniotic shunting (a twin–twin transfusion syndrome with Laser ablation of vessels), the treatment of fetal bladder obstructions, an aortic or pulmonary valvuloplasty (opening the aortic or pulmonary fetal heart valves to allow blood flow) (6), an atrial septostomy (opening the inter-atrial septum of the fetal heart to allow unrestricted blood flow between the atriums), the surgical treatment of a congenital diaphragmatic hernia by a Balloon tracheal occlusion and the treatment of spina bifida with a closure of the malformation entered clinical practice. Epidural anesthesia, the new standardized criteria of fetal heart rate analysis, the advent of the STAN, the analysis of scalp pH contributed to the improvement of perinatal prognosis. Techniques of embolization contributed to a less aggressive, but effective, treatment of post-partum hemorrhage, thus saving lives and sparing uteri. With such a therapeutic arsenal, there should be no more fatality. However questions arise, among them “should we go back to a more physiological or natural practice of obstetrics?” or “why the rate of cesarean sections increased, and all this technology is available?” Even though we cannot compromise progresses, the question is to be debated. In both gynecology and obstetrics, optimal training of the nurses, midwifes, and residents is essential. In these fields, specific simulators are ideal tools to start, maintain, and develop skills. Finally, internet and other media are obvious tools to enhance knowledge for patients, doctors, midwives, and nurses.

Objective: In this article, we present two case reports. The first case was a malignant germ cell tumor of the right ovary in a 23 year old female and a case of a bilateral undifferentiated granulosa cell tumor in a 71 year old female. The aim of these reports is to illustrate the interest of the immuno-histochemical analysis to define the correct diagnosis, to better classify these ovarian tumours and improve their management. Methods: This study we report two cases. The first case concern a 23 years old woman (A) with a mixed germ cell tumour of the right ovary (dysgerminoma (75%), yolk sac tumour (20%), and a mature teratoma (5%), and the second case (B) a bilateral non differentiated and necrotic granulosa cell tumour of both ovaries concerning a 71 year old patient. The staging system used was according to both classifications, the one of International Federation of Gynaecology and Obstetrics (FIGO) 1987 for ovarian cancer, and the one of TNM code 2009. Results: The immunostaining establish the malignancy and the immunochemistry contribute to confirm effectively the right diagnosis (Table 2 and 3). Conclusion: An immuno-histochemical analysis is mandatory for the choice of chemotherapy to obtain the better response of the disease and improve the survival prognosis. The efficiency of the chemotherapy authorizes a conservative surgery including a unilateral salpingo-oophorectomy preserving fertility (A). Concerning the non-dysgerminoma tumour (B), and after a surgical staging and debulking, chemotherapy was recommended. The type of tumour and its histological feature conditioned the choice of treatment. The benefit of the immunohistological analysis in this case allowed the right adjuvant treatment. Key words: germ cell tumours, dysgerminoma, teratoma, yolk sac, ovarian cell tumour, and immunohistochemistry.

Placenta percreta retention within the scar of a previous caesarean section is rare. We report here one of these cases treated successfully by laparoscopy, with uterine repair. Different therapeutic options are described.

Trisomy 21 is the most frequent genetic cause of cognitive impairment. To assess the perturbations of gene expression in trisomy 21, and to eliminate the noise of genomic variability, we studied the transcriptome of fetal fibroblasts from a pair of monozygotic twins discordant for trisomy 21. Here we show that the differential expression between the twins is organized in domains along all chromosomes that are either upregulated or downregulated. These gene expression dysregulation domains (GEDDs) can be defined by the expression level of their gene content, and are well conserved in induced pluripotent stem cells derived from the twins' fibroblasts. Comparison of the transcriptome of the Ts65Dn mouse model of Down's syndrome and normal littermate mouse fibroblasts also showed GEDDs along the mouse chromosomes that were syntenic in human. The GEDDs correlate with the lamina-associated (LADs) and replication domains of mammalian cells. The overall position of LADs was not altered in trisomic cells; however, the H3K4me3 profile of the trisomic fibroblasts was modified and accurately followed the GEDD pattern. These results indicate that the nuclear compartments of trisomic cells undergo modifications of the chromatin environment influencing the overall transcriptome, and that GEDDs may therefore contribute to some trisomy 21 phenotypes.

The trophoblast, which is derived from the extraembryonic trophectoderm, is the first cells differentiated from the pre-implantation mammalian embryos. In this regard, two differentiation pathways are critical for the survival and development of the embryo in utero. In the first one, cytotrophoblasts (CTBs) fuse into a layer of syncytiotrophoblasts that covers the chorionic villi. These cells are responsible for hormone production, nutrient and gas exchanges between the mother and the developing fetus. In the second one, CTBs in the anchoring chorionic villi invade the uterus and its blood vessels establishing the maternal-fetal interface (Red-Horse et al., 2004; Roberts and Fisher, 2011). Dysfunction of these differentiation pathways is associated with a wide range of human pregnancy complications such as infertility, preterm birth, pre-eclampsia, intrauterine growth restriction and aneuploidies (Red-Horse et al., 2004). To date, most of the insights on placental trophoblast development have been gained through transgenic mouse models and mouse trophoblast stem cells with placental defects (Kunath et al., 2004). Despite similarities to human phenotype, mouse models have several drawbacks and cannot integrate the specificities of human placentation. To overcome the unfeasibility of performing in vivo experimentation in humans, cell lines derived from human choriocarcinoma or virally transformed human trophoblast cell lines are widely used to investigate the mechanisms involved in trophoblast differentiation (Hannan et al., 2010). However, general limitations of using cell lines exist such as their genetic background and the potential changes acquired after their transformation and during their establishment in culture. Alternatively, primary trophoblast cultures have been derived from human placentas at the first trimester or at term of pregnancy. While these cells might be an excellent model to study trophoblast differentiation, they are often difficult to obtain and do not proliferate in culture. In addition, these cells are already committed to the trophoblast lineage and therefore are not amenable to study early trophoblast development. The discovery that human pluripotent stem cells (hPSCs) can be differentiated into trophoblast cells through bone morphogenetic proteins (BMPs) (Xu et al., 2002) has opened up a new field of investigation in human trophoblast developmental biology and disease (Ezashi et al., 2012; Golos et al., 2013). Thomson's group was the first showing trophoblast differentiation of human embryonic stem cells (hESCs) following BMP4 treatment (Xu et al., 2002). Then, the same group has shown that in the presence of high concentrations of FGF2, BMP4 treatment results in the differentiation of hESCs into mesendoderm rather than trophoblast. They demonstrated in particular that FGF signaling switches BMP4-induced differentiation of hESCs into mesendoderm via the maintenance of NANOG expression through MEK/ERK pathway (Yu et al., 2011). Thereafter, other studies have emphasized the need of dual inhibition of FGF signaling (with PD173074 or SU5402) and TGF-β/ACTIVIN/NODAL signaling (with SB431542 or A83-01) to support trophoblast induction specifically (Sudheer et al., 2012; Amita et al., 2013). Under these conditions, BMP4-induced differentiation leads to the emergence of both chorionic gonadotrophin β-secreting syncytiotrophoblasts with a villous CTB phenotype and HLA-G+ trophoblast cells with invasive properties as the extravillous CTBs (Marchand et al., 2011; Sudheer et al., 2012; Amita et al., 2013; Li et al., 2013). Furthermore, by their ability to recapitulate the early and late steps of trophoblast development, hESCs should become valuable tools to study the cellular and molecular mechanisms involved in trophoblast cell maintenance, specification and differentiation. Since 2007, the generation of human induced pluripotent stem cells (hiPSCs) has allowed researchers to study the development and/or progression of several pathologies including neurological diseases [reviewed in Hibaoui and Feki (2012)]. It is undoubted that similar approaches using patient-specific iPSCs will offer the opportunity to reproduce normal and pathological trophoblast development. Moreover, hiPSCs exhibited additional advantages compared to hESCs. First, hiPSCs do not require the use of human embryos, which makes their use in basic research less ethically controversial. They are also technically easier to obtain. Last but not least, they offer the opportunity to generate iPSC lines specific for patients with pregnancy complications. Of note, the recent discovery that iPSCs generated by in vivo reprogramming of adult mouse cells efficiently differentiate into the three germ layers and extraembryonic tissue (Abad et al., 2013), may improve our understanding of extraembryonic lineage development and placenta-associated disorders. In conclusion, while further studies are needed to improve the efficiency of hPSC differentiation into trophoblast cells, we believe that this technology provides a promising alternative model to study trophoblast development. In fact, understanding and elucidating the mechanisms regulating human trophoblast development using hPSCs is critical not only in basic research but also in clinical medicine. It could provide novel insights into blastocyst formation, implantation and placentation processes in humans. Thus, patient-specific hPSCs will also open up new exciting avenues for the identification of genetic and biochemical markers that could be useful for the prediction and management of pregnancy complications such as pre-eclampsia, intrauterine growth restriction and aneuploidies. The coming years will tell whether these cells fulfil their promise.

International audience; Sub-threshold operation of circuits becomes more and more attractive due to the ultra-low power consumption. Static Random Access Memory (SRAM) faces an important limitation in read access time that prevents high frequency operation and the possible applications. The read access time under ultra-low voltage (ULV) operation is mainly dictated by the read current of the SRAM bit cell and the bit line effective capacitance. The full swing sensing is a practical approach to circumvent the poor performances of sense amplifiers (SA) under ULV operation. This paper details first the optimization of a differential voltage-sense amplifier under ULV for SRAMs with differential bit lines. Second an unbalanced voltage-sense amplifier is presented for single-ended reading under ULV. Both circuits exploit the benefit of 28nm FDSOI and back biasing technique to improve SAs' performances, namely the delay. Both ultra-wide voltage-range SAs achieve satisfying operation down to 280mV power supply. Simulation results are presented regarding a 1K×32 L1 cache test chip to be fabricated in 28FDSOI technology.

The aim of the present study was to evaluate fertility outcome after laparoscopic microsurgical tubal anastomosis (one-stitch technique). We carried out a retrospective study of 51 patients seeking reversal of previous tubal sterilization. The mean age of the patients was 33.2 years (range: 30-44.5 years). Surgery lasted 122 minutes on average, (60-204 minutes). Mean time to pregnancy was 8.2 months (4-32 months) and the rate of pregnancy obtained was 56.2%. Twenty seven patients became pregnant and 12 had ectopic pregnancies; of the 15 intrauterine pregnancies, 3 miscarried spontaneously before 9 weeks. Our study observed valuable qualities of feasibility, simplicity and rapidity of the “one-stitch technique” in laparoscopic tubal anastomosis surgery, with related good rates of intrauterine pregnancy. The rate of pregnancy obtained is fairly similar to the rates reported in the literature with the same technique. However, the high proportion of ectopic pregnancies makes us believing that laparoscopic procedures using more than one single stitch should be preferred.