Cryoconservation ovarienne : Étude de la folliculogenèse in vitro chez l'ovin et chez<br />l'humain

Culture of ovarian cortex can be an alternative to autograft after ovarian cryopreservation. To improve follicular survival after freezing we evaluated the influence of the sheep cortex width; 0.5 vs 1mm, of the culture support and the cryoprotector DMSO 2M vs PROH 1.5M + sucrose 0.3M. The 1mm tissue width on millicell filter with the DMSO 2M freezing protocol displayed better results with 39% follicular survival on the 4th day of culture. On human and sheep fresh tissue, we explored a potential expression deficit in culture of Gdf9, Bmp15 and their receptors; this would explain the blocking of follicular growth in vitro. In both cases the level of ARNm and proteins Gdf9 and Bmp15 is maintained and so is the protein formula of their BmprIA, BmprIB, BmprII receptors. This study shows how survival in culture after cryopreservation can be optimised, the reason of the growth blocking remains to be determinedLa culture de cortex ovarien se pose comme une alternative à l'autogreffe après autoconservation ovarienne. Pour améliorer la survie folliculaire après congélation, nous avons évalué chez la brebis l'influence de l'épaisseur de cortex ; 0,5 vs 1mm, du support de culture et du cryoprotecteur DMSO 2M vs PROH 1,5M+sucrose 0,3M. L'épaisseur de tissu de 1mm sur filtre millicell avec le protocole de congélation DMSO 2M a montré les meilleurs résultats avec 39% de survie folliculaire au 4e jour de culture. Sur tissu frais de brebis et humain, nous avons exploré un éventuel déficit d'expression en culture de Gdf9, de Bmp15 et de leur récepteurs qui expliquerait le blocage de croissance folliculaire in vitro. Dans les deux modèles la présence des ARNm et des protéines de Gdf9 et Bmp15 est maintenue et l'expression protéique de leurs récepteurs BmprIA, BmprIB, BmprII est conservée. Ce travail montre que la survie en culture après cryoconservation peut être optimisée, la cause du blocage de croissance reste à défini

Autoconservation ovarienne (pratiques de l'autoconservation ovarienne au sein du département de Médecine de la Reproduction : études expérimentales de folliculogénèse in vitro chez l'ovin)

LYON1-BU Santé (693882101) / SudocPARIS-BIUM (751062103) / SudocSudocFranceF

Ruptura prematura de membranas: factores asociados morbimortalidad materna y perinatal en el hospital regional del Amazonas Iquitos: marzo - octubre 1991

Being Premature Rupture of Membranes an obstetric problem quite often leading to increased maternal and perinatal morbidity; this prospective comparative study was conducted at the Regional Hospital Amazon - Iquitos, from March to October 1991 to determine the incidence and identify factors associated with RPH also determine the latency that causes increased morbidity and mortality in the mother and the newborn. Of a total of 1,364 births, 215 RPM found pregnant representing an incidence of 15.76 \, was selected 368 pregnant women between 20-41 weeks who met protocol requirements and half of them represented the experimental group (184 ) with RPM, l other half (184) was the Control group without RPM. The rate of perinatal mortality was 37.20x1000 NV RPM, while the overall perinatal mortality rate was 23.46 X 1000 NV Factors associated with the RPH were; Primigravida adolescents, lack of prenatal and leucorrhea control. The latency period that produces increased maternal and perinatal morbidity is greater than 24 hours.Siendo la Rotura Prematura de Membranas un problema obstétrico de relativa frecuencia que conlleva un aumento de la morbilidad materna y perinatal; se realizó el presente estudio de carácter prospectivo comparativo en el Hospital Regional del Amazonas - Iquitos, entre Marzo-Octubre de 1991 para conocer la incidencia e identificar los factores asociados a la RPH, además determinar el periodo de latencia que produce mayor morbimortalidad en la madre y el recién nacido. De un total de 1,364 partos, se encontró 215 gestantes con RPM representando una incidencia de 15.76\, Se seleccionó a 368 gestantes entre 20-41 semanas, que cumplieron con los requisitos del protocolo, así la mitad de ellos representó el grupo experimental (184) con RPM, y l otra mitad (184) constituyó el Grupo control, sin RPM. La tasa de mortalidad perinatal en la RPM fue de 37.20x1000 N.V., mientras que la tasa de mortalidad perinatal global fue de 23.46 X 1000 N.V. Los factores asociados a la RPH fueron; Primigesta adolescentes, ausencia de control prenatal y leucorreas. El periodo de latencia que produce mayor morbilidad materna y perinatal es mayor de 24 horas.Tesi

Cryoconservation ovarienne (étude de la folliculogenèse in vitro chez l ovin et chez l humain)

Culture of ovarian cortex can be an alternative to autograft after ovarian cryopreservation. To improve follicular survival after freezing we evaluated the influence of the sheep cortex width; 0.5 vs 1mm, of the culture support and the cryoprotector DMSO 2M vs PROH 1.5M + sucrose 0.3M. The 1mm tissue width on millicell filter with the 2DMSO 2M freezing protocol displayed better results with 39% follicular survival on the 4th day of culture. On fresh human and sheep tissue, we explored a potential expression deficit in culture of Gdf9, Bmp15 and their receptors; this would explain the blocking of follicular growth in vitro. In both cases the level of ARNm and proteins Gdf9 and Bmp15 is maintained and so is the protein formula of their BmprIA, BmprIB, BmprII receptors. This study shows how survival in culture after cryopreservation can be optimised; the reason of the growth blocking remains to be determinedLa culture de cortex ovarien se pose comme une alternative à l autogreffe après autoconservation ovarienne. Pour améliorer la survie folliculaire après congélation, nous avons évalué chez la brebis l influence de l épaisseur de cortex ; 0,5 vs 1mm, du support de culture et du cryoprotecteur DMSO 2M vs PROH 1,5M+sucrose 0,3M. L épaisseur de tissu de 1mm sur filtre millicell avec le protocole de congélation DMSO 2M a montré les meilleurs résultats avec 39% de survie folliculaire au 4e jour de culture. Sur tissu frais de brebis et humain, nous avons exploré un éventuel déficit d expression en culture de Gdf9, de Bmp15 et de leurs récepteurs qui expliquerait le blocage de croissance folliculaire in vitro. Dans les deux modèles la présence des ARNm et des protéines de Gdf9 et Bmp15 est maintenue et l expression protéique de leurs récepteurs BmprIA, BmprIB, BmprII est conservée. Ce travail montre que la survie en culture après cryoconservation peut être optimisée, la cause du blocage de croissance reste à définirLYON1-BU.Sciences (692662101) / SudocSudocFranceF

Impact of Nanoparticles on Male Fertility: What Do We Really Know? A Systematic Review

International audienceThe real impact of nanoparticles on male fertility is evaluated after a careful analysis of the available literature. The first part reviews animal models to understand the testicular biodistribution and biopersistence of nanoparticles, while the second part evaluates their in vitro and in vivo biotoxicity. Our main findings suggest that nanoparticles are generally able to reach the testicle in small quantities where they persist for several months, regardless of the route of exposure. However, there is not enough evidence that they can cross the blood-testis barrier. Of note, the majority of nanoparticles have low direct toxicity to the testis, but there are indications that some might act as endocrine disruptors. Overall, the impact on spermatogenesis in adults is generally weak and reversible, but exceptions exist and merit increased attention. Finally, we comment on several methodological or analytical biases which have led some studies to exaggerate the reprotoxicity of nanoparticles. In the future, rigorous clinical studies in tandem with mechanistic studies are needed to elucidate the real risk posed by nanoparticles on male fertility

Impact of Nanoparticles on Male Fertility: What Do We Really Know? A Systematic Review

The real impact of nanoparticles on male fertility is evaluated after a careful analysis of the available literature. The first part reviews animal models to understand the testicular biodistribution and biopersistence of nanoparticles, while the second part evaluates their in vitro and in vivo biotoxicity. Our main findings suggest that nanoparticles are generally able to reach the testicle in small quantities where they persist for several months, regardless of the route of exposure. However, there is not enough evidence that they can cross the blood–testis barrier. Of note, the majority of nanoparticles have low direct toxicity to the testis, but there are indications that some might act as endocrine disruptors. Overall, the impact on spermatogenesis in adults is generally weak and reversible, but exceptions exist and merit increased attention. Finally, we comment on several methodological or analytical biases which have led some studies to exaggerate the reprotoxicity of nanoparticles. In the future, rigorous clinical studies in tandem with mechanistic studies are needed to elucidate the real risk posed by nanoparticles on male fertility

Follicular growth in vitro: Detection of growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15) during in vitro culture of ovine cortical slices

International audiencePrimordial follicles from different mammal species can survive and enter the growth phase in vitro but do not develop beyond the primary stage. The hypothesis was that, in sheep, in vitro follicular growth is arrested because of a lack of secretion of GDF9 and/or BMP15. Cortical slices of 0.3-0.5 mm thickness issued from 5- to 6-month-old lambs were cultured for 15 days. The pieces were fixed on days 0, 2, 4, 7, 10, and 15 of culture. Follicle morphology, RT-PCR exploration of GDF9 and BMP15 mRNA, immunolhistochemical location of their proteins and their receptor BMPRIB and BMPRII were assessed at different time of culture. The mean percentage of primordial follicles decreased from 58.6% (day 0) to 13.4% (day 15) (P < 0.01), whereas that of primary follicles increased from 3.2% (day 0) to 31.5% on day 4 (P < 0.01), then remained stable until day 15 (35.6%). The percentage of atretic follicles increased from 14.7% (day 0) to 27.1% (day 15) (P < 0.05). A few secondary follicles were observed on days 4 and 10, representing 1.0%, and 2.1% of the total number of follicles. GDF9 and BMP15 mRNAs were detected from harvesting (day 0) up to day 15 following culture. At the same time, positive immunoreactions for GDF9, BMP15 and for BMPRIB and BMPRII were also found in oocyte cytoplasm. In conclusion, expression of GDF9, BMP15 and their receptors BMPRIB and BMPRII are detected during in vitro culture of ovine cortical slices

Methylation profile of KCNQ1OT1 gene in in vitro maturated human oocytes and non growing embryos

National audienc

Ex vivo detection and quantification of gold nanoparticles in human seminal and follicular fluids

International audienceIncreasing consumption of engineered nanoparticles and occupational exposure to novel, ultrafine airborne particles during the last decades has coincided with deterioration of sperm parameters and delayed fecundity. In order to prevent possible adverse health effects and ensure a sustainable growth for the nanoparticle industry, the ability to investigate the nanosized, mineralogical load of human reproductive systems is becoming a real clinical need. Toward this goal, the current study proposes two methods for the detection and quantification of engineered nanoparticles in human follicular and seminal fluid, developed with the use of well-defined 60 nm Au particles. Despite the complexity of these biological fluids, simple physical and chemical treatments allow for the precise quantification of more than 50 and 70% wt of the spiked Au nanoparticles at low μg ml−1 levels in follicular and seminal fluids, respectively. The use of electron microscopy for the detailed observation of the detected analytes is also enabled. The proposed method is applied on a small patient cohort in order to demonstrate its clinical applicability by exploring the differences in the metal and particulate content between patients with normal and low sperm count

Toward the Improvement of Silicon-Based Composite Electrodes via an In-Situ Si@C-Graphene Composite Synthesis for Li-Ion Battery Applications

International audienceUsing Si as anode materials for Li-ion batteries remain challenging due to its morphological evolution and SEI modification upon cycling. The present work aims at developing a composite consisting of carbon-coated Si nanoparticles (Si@C NPs) intimately embedded in a three-dimensional (3D) graphene hydrogel (GHG) architecture to stabilize Si inside LiB electrodes. Instead of simply mixing both components, the novelty of the synthesis procedure lies in the in situ hydrothermal process, which was shown to successfully yield graphene oxide reduction, 3D graphene assembly production, and homogeneous distribution of Si@C NPs in the GHG matrix. Electrochemical characterizations in half-cells, on electrodes not containing additional conductive additive, revealed the importance of the protective C shell to achieve high specific capacity (up to 2200 mAh.g−1), along with good stability (200 cycles with an average Ceff > 99%). These performances are far superior to that of electrodes made with non-C-coated Si NPs or prepared by mixing both components. These observations highlight the synergetic effects of C shell on Si NPs, and of the single-step in situ preparation that enables the yield of a Si@C-GHG hybrid composite with physicochemical, structural, and morphological properties promoting sample conductivity and Li-ion diffusion pathways