Circulating microRNAs in follicular fluid, powerful tools to explore in vitro fertilization process.

Circulating or "extracellular" microRNAs (miRNAs) detected in biological fluids, could be used as potential diagnostic and prognostic biomarkers of several disease, such as cancer, gynecological and pregnancy disorders. However, their contributions in female infertility and in vitro fertilization (IVF) remain unknown. This study investigated the expression profiles of five circulating miRNAs (let-7b, miR-29a, miR-30a, miR-140 and miR-320a) in human follicular fluid from 91 women with normal ovarian reserve and 30 with polycystic ovary syndrome (PCOS) and their ability to predict IVF outcomes. The combination of FF miR-30a, miR-140 and let-7b expression levels discriminated between PCOS and normal ovarian reserve with a specificity of 83.8% and a sensitivity of 70% (area under the ROC curve, AUC = 0.83 [0.73-0.92]; p < 0.0001). FF samples related to low number of mature oocytes (≤2) contained significant less miR-320a levels than those related to a number of mature oocytes >2 (p = 0.04). Moreover, FF let-7b predicted the development of expanded blastocysts with 70% sensitivity and 64.3% specificity (AUC = 0.67 [0.54-0.79]; p = 0.02) and FF miR-29a potential to predict clinical pregnancy outcome reached 0.68 [0.55-0.79] with a sensitivity of 83.3% and a specificity of 53.5% (p = 0.01). Therefore, these miRNAs could provide new helpful biomarkers to facilitate personalized medical care during IVF.

[Circulating nucleic acids and infertility]. / Les acides nucléiques circulants et infertilité.

Circulating nucleic acids (cell-free DNA and microRNAs) have for particularity to be easily detectable in the biological fluids of the body. Therefore, they constitute biomarkers of interest in female and male infertility care. Indeed, in female, they can be used to detect ovarian reserve disorders (polycystic ovary syndrome and low functional ovarian reserve) as well as to assess follicular microenvironment quality. Moreover, in men, their expression levels can vary in case of spermatogenesis abnormalities. Finally, circulating nucleic acids have also the ability to predict successfully the quality of in vitro embryo development. Their multiple contributions during assisted reproductive technology (ART) make of them biomarkers of interest, for the development of new diagnostic and/or prognostic tests, applied to our specialty. Circulating nucleic acids would so offer the possibility of personalized medical care for infertile couples in ART.

Sperm global DNA methylation level: association with semen parameters and genome integrity.

Sperm DNA methylation abnormalities have been detected in oligozoospermic men. However, the association between sperm DNA methylation defects, sperm parameters and sperm DNA, and chromatin integrity remains poorly understood. This study was designed to clarify this issue. We recruited a cohort of 92 men (62 normozoospermic and 30 oligoasthenozoospermic) presenting for infertility evaluation during a 1-year period. Sperm global DNA methylation was evaluated by an ELISA-like method, DNA fragmentation was evaluated by flow cytometry-based terminal transferase dUTP nick end-labeling (TUNEL) assay (reported as DNA fragmentation index or DFI), and sperm denaturation was evaluated by aniline blue staining (reported as sperm denaturation index or SDI, a marker of chromatin compaction). We found a significant positive association between sperm global DNA methylation level and conventional sperm parameters (sperm concentration and motility), supported by the results of methylation analysis on H19-DMR. We also identified significant inverse relationships between sperm global DNA methylation, and, both DFI and SDI. However, sperm global DNA methylation level was not related to sperm vitality or morphology. Our findings suggest that global sperm DNA methylation levels are related to conventional sperm parameters, as well as, sperm chromatin and DNA integrity.

Cell-free nucleic acids as non-invasive biomarkers of gynecological cancers, ovarian, endometrial and obstetric disorders and fetal aneuploidy.

BACKGROUND: Proper folliculogenesis is fundamental to obtain a competent oocyte that, once fertilized, can support the acquisition of embryo developmental competence and pregnancy. MicroRNAs (miRNAs) are crucial regulators of folliculogenesis, which are expressed in the cumulus-oocyte complex and in granulosa cells and some can also be found in the bloodstream. These circulating miRNAs are intensively studied and used as diagnostic/prognostic markers of many diseases, including gynecological and pregnancy disorders. In addition, serum contains small amounts of cell-free DNA (cfDNA), presumably resulting from the release of genetic material from apoptotic/necrotic cells. The quantification of nucleic acids in serum samples could be used as a diagnostic tool for female infertility. METHODS: An overview of the published literature on miRNAs, and particularly on the use of circulating miRNAs and cfDNA as non-invasive biomarkers of gynecological diseases, was performed (up to January 2014). RESULTS: In the past decade, cell-free nucleic acids have been studied for potential use as biomarkers in many diseases, particularly in gynecological cancers, ovarian and endometrial disorders, as well as in pregnancy-related pathologies and fetal aneuploidy. The data strongly suggest that the concentration of cell-free nucleic acids in serum from IVF patients or in embryo culture medium could be related to the ovarian hormone status and embryo quality, respectively, and be used as a non-invasive biomarker of IVF outcome. CONCLUSIONS: The profiling of circulating nucleic acids, such as miRNAs and cfDNA, opens new perspectives for the diagnosis/prognosis of ovarian disorders and for the prediction of IVF outcomes, namely (embryo quality and pregnancy).

[Oxidative stress and fertility: false evidence and bad recipes]. / Stress oxydant et fertilité : fausses évidences et mauvaises recettes.

Worldwide statistics agree that at least one out of six couples has fertility problems. If the male gamete is the origin of this problem, it is generally admitted that the oxidative stress is involved. Modern life has obviously increased fertility problems through pesticides, xenoestrogenes, endocrine disrupting chemicals involved in plastic technology such as polychlorinated bisphenyls, bisphenol A, phthalates and alkylphenols… and other cosmetic additives. An important part of these compounds increases oxidative stress, at least in part. Oxidative stress is more than probably at the origin or recurrent increasing pathologies such as endometriosis. If the oocyte is theoretically able to repair oxidative stress linked decays such as DNA fragmentation and oxidation of bases, its capacity is finite and decreasing with age. In order to decrease DNA repair charge, reducing or even avoiding the generation of DNA damages related to reactive oxygen species through consumption of antioxidants compounds is often tempting: however Reasons will be provided to break from current treatments given haphazardly in the population in the age of reproduction, as well as the potential risks of over-exposure. Furthermore recommended treatments, in relation with the new concepts in oxidative stress, will be specified.

[Teratozoospermia, myth or reality? A 10-years retrospective study on 101404 consecutive sperm samples]. / Tératozoospermie: mythe ou réalité? Étude d'une cohorte de 101 404 examens de sperme.

OBJECTIVES: Several studies have reported a time-related decline in semen quality. In this context, 101,404 spermograms-spermocytograms performed in a single andrology laboratory from January 2000 to December 2009 were investigated retrospectively. Variations of sperm parameters were analyzed over the years. PATIENTS AND METHODS: For each semen sample, age at semen collection, duration of sexual abstinence, volume of seminal fluid, pH, concentration of spermatozoa, percentages of overall motile and progressive motile sperm, percentage of morphologically abnormal spermatozoa (according to David's classification) and amplitude of lateral head displacement (ALH) were analyzed. For each parameter, the mean value per year was determined. To examine trends over time the statistical tests used were analyses of variance and correlation studies. RESULTS: Data showed an increasing age of patients from 36.5 years in 2000 to 37.2 years in 2009. The semen concentration was 63.5 millions/mL in 2000 and 63.6 millions/mL in 2009 with a slight fall from 2001 to 2008. The vitality continuously increased from 67.8% in 2000 to 73.5% in 2009 as well as the overall motility (from 37.7% to 39.2%) and the progressive motility (from 34.6% to 36.2%). The percentage of morphologically abnormal spermatozoa continuously increased from 64.5% to 84.2% (r=0.43, P<0.001). DISCUSSION AND CONCLUSION: Contrary to a variety of works, our very large study of semen analysis did not show a real decline in semen WHO parameters during a 10-year period, except for the percentage of morphologically abnormal spermatozoa, which dramatically increased over the years.

[Teratozoospermia at the time of intracytoplasmic morphologically selected sperm injection (IMSI)]. / La tératozoospermie à l'heure de l'intracytoplasmic morphologically selected sperm injection (IMSI).

Until now, the morphological sperm analysis (spermocytogram) allows to define sperm normality, but the relationship between sperm morphology and fertility is not yet assessed. Although several studies do not report any relationship between abnormal sperm morphology and ICSI results, nevertheless, the success rate of ICSI sems to be dependent on injected sperm morphological aspect. Detailed morphological sperm examination (especially sperm head) at high magnification (from x 6600 to x 12500) (MSOME) in real time allows to select the best spermatozoa before oocyte injection (IMSI). In some cases, implantation and ongoing pregnancy rates were improved with this sperm selection method. Ultramorphologic criteria were established and the most predictive factor of sperm quality is the presence of vacuoles in the sperm head. Those vacuoles appear to be related to DNA damage (fragmentation and/or denaturation) and affect embryo development. To standardize those observations, several authors tried to establish sperm MSOME classifications in order to be used in routine and to replace the conventional spermocytogram in the next future.

[Indications for IMSI]. / Les indications de l'IMSI.

ICSI failures (essentially fertilization and implantation failures) can be due to the quality of the oocytes, the spermatozoon injected and its manipulation, uterine factors and paternal factors, such as DNA fragmentation of the spermatozoa. Pre-selection of the spermatozoon is the crucial phase for ensuring successful ICSI. The characteristics of the sperm must be checked under the microscope at a magnification of x 6600 as certain abnormalities, in particular nuclear vacuoles, cannot be seen at x 300. In our centre, we conducted a study on 72 patients which showed that with more than 30% of fragmented DNA, neither implantation or birth was achieved using ICSI. Performing IMSI on the same patients allowed us to obtain implantation rates of 17.4% with 30 to 40 % of fragmented DNA and of 33.3% above 40%. Births were achieved in 17.4% and 28.6% of cases respectively. IMSI has the advantage of allowing extremely careful selection of the spermatozoon microinjected, so that it is of normal shape, with the least vacuoles possible and a low DNA fragmentation rate. It is, however, a complicated technique that cannot be routinely performed.

Análisis descriptivo de la patología neurológica en una unidad de cuidados intensivos pediátricos de referencia regional / Descriptive analysis of neurological disorders in the pediatric intensive care unit of a regional reference hospital

OBJETIVO: El objetivo de este trabajo ha sido conocer y analizar los problemas neurológicos de la UCI Pediátríca (UCIP) del Hospital Infantil Miguel Servet de Zaragoza. De la base de datos de la sección de neuropediatría, que incluye a todos los niños ingresados en UCIP con problemas neurológicos, se han estudiado los niños ingresados en UCIP desde el 15 de mayo de 1990 hasta el 31 de diciembre de 1999. RESULTADOS: De los 4.507 niños valorados por la sección de neuropediatría durante el período de estudio de 9 años y 6 meses, 591 han ingresado al menos una vez en la UCIP, con un total de 711 ingresos (32 por ciento de los 2.198 ingresos en UCIP). Fallecieron en UCIP 60 niños con problemas neurológicos. Los motivos de ingreso más frecuentes han sido: 212 traumatismos craneoencefálicos (TCE) (30 por ciento), 127 postoperatorios neuroquirúrgicos (18 por ciento), 110 convulsiones (15 por ciento), 72 encefalopatías agudas (10 por ciento), 44 insuficiencias respiratorias (6,1 por ciento) y 28 accidentes diferentes al TCE (4 por ciento). CONCLUSIÓN: La patología neurológica representa una amplia parte de la actividad en nuestra unidad de cuidados intensivos pediátricos. Algunos problemas frecuentes como la encefalopatía aguda o las convulsiones exigen un importante esfuerzo diagnóstico, puesto que pueden ser manifestación de diversos problemas neurológicos y extraneurológicos. Algunos de los problemas neurológicos que requieren cuidados intensivos, como los traumatismos craneoencefálicos y diversas encefalopatías agudas, asocian elevadas morbilidad y mortalidad (AU)