Effects of Simulated Microgravity In Vitro on Human Metaphase II Oocytes: An Electron Microscopy-Based Study.

The Gravity Force to which living beings are subjected on Earth rules the functionality of most biological processes in many tissues. It has been reported that a situation of Microgravity (such as that occurring in space) causes negative effects on living beings. Astronauts returning from space shuttle missions or from the International Space Station have been diagnosed with various health problems, such as bone demineralization, muscle atrophy, cardiovascular deconditioning, and vestibular and sensory imbalance, including impaired visual acuity, altered metabolic and nutritional status, and immune system dysregulation. Microgravity has profound effects also on reproductive functions. Female astronauts, in fact, suppress their cycles during space travels, and effects at the cellular level in the early embryo development and on female gamete maturation have also been observed. The opportunities to use space flights to study the effects of gravity variations are limited because of the high costs and lack of repeatability of the experiments. For these reasons, the use of microgravity simulators for studying, at the cellular level, the effects, such as those, obtained during/after a spatial trip, are developed to confirm that these models can be used in the study of body responses under conditions different from those found in a unitary Gravity environment (1 g). In view of this, this study aimed to investigate in vitro the effects of simulated microgravity on the ultrastructural features of human metaphase II oocytes using a Random Positioning Machine (RPM). We demonstrated for the first time, by Transmission Electron Microscopy analysis, that microgravity might compromise oocyte quality by affecting not only the localization of mitochondria and cortical granules due to a possible alteration of the cytoskeleton but also the function of mitochondria and endoplasmic reticulum since in RPM oocytes we observed a switch in the morphology of smooth endoplasmic reticulum (SER) and associated mitochondria from mitochondria-SER aggregates to mitochondria-vesicle complexes. We concluded that microgravity might negatively affect oocyte quality by interfering in vitro with the normal sequence of morphodynamic events essential for acquiring and maintaining a proper competence to fertilization in human oocytes.

Could art cycles have a detrimental effect on ovarian reserve? A retrospective case control study.

BACKGROUND: Even if it is supposed damage of repeated ART (assisted reproductive technology) cycles on oocyte pool, there is still no evidence in literature. Aim of the study is to investigate whether infertile women who undergo to several ART cycles can show a lower ovarian reserve measured by AMH (Anti-Mullerian hormone) levels. METHODS: The study includes 282 infertile women, between 18 and 42 years, and allocated into two groups: 159 women previously submitted to two or more ART cycles (group A) and 123 women never submitted naïve to-ART cycles (group B). We tested whether AMH, FSH, LH and E2 levels were significantly different between the two groups, stratifying according to age. RESULTS: Regardless to the age ranges bands, the AMH in group A was statistically significant lower than in group B with a statistical significance (P=0.047). In particular women aged over 35 previously submitted to one or more ART cycles showed lower AMH levels, than those paired with age, which had never been treated with ART. CONCLUSIONS: Despite the limitations of the study, our data demonstrate a reduced AMH levels in women aged over 35 previously submitted to two or more repeated ART-cycles compared to patients never treated before. The strength of this study is the actuality of the topic that has not been discussed before in detail.

Revealing the secret life of pre-implantation embryos by time-lapse monitoring: A review.

High implantation success following in vitro fertilization cycles are achieved via the transfer of embryos with the highest developmental competence. Multiple pregnancies as a result of the transfer of several embryos per cycle accompany with various complication. Thus, single-embryo transfer (SET) is the preferred practice in assisted reproductive technique (ART) treatment. In order to improve the pregnancy rate for SET, embryologists need reliable biomarkers to aid their selection of embryos with the highest developmental potential. Time-lapse technology is a noninvasive alternative conventional microscopic assessment. It provides uninterrupted and continues the survey of embryo development to transfer day. Today, there are four time-lapse systems that are commercially available for ART centers. In world and Iran, the first time lapse babies were born in 2010 and 2015, respectively, conceived by SET. Here, we review the use of time-lapse monitoring in the observation of embryogenesis as well as its role in SET. Although, the findings from our review support common use of time-lapse monitoring in ART centers; but, future large studies assessing this system in well-designed trials are necessary.

Successful use of "Hour 1" transfer of sperm microinjected oocytes in natural cycles of poor responder women: a prospective randomized controlled pilot study.

The aim of the study is to demonstrate the successful use of "Hour 1" transfer of sperm microinjected oocytes in natural cycles of poor responder women. Seventy women were selected from 92 consecutive poor responders undergoing intracytoplasmatic sperm injection (ICSI) in a natural cycle at our Sterility and Assisted Reproduction Unit from September 2009 to July 2013, and randomly distributed in two homogeneous groups: Group A or B. Women in Group A (35) underwent transfer within 1 h after ICSI; Group B (35, control group) underwent transfer 3 days after ICSI. In Group A, seven clinical pregnancy and one miscarriage occurred; in Group B, eight clinical pregnancies and two miscarriages were observed. Pregnancy, miscarriage, term pregnancy and overall live birth rates' difference between the two groups was not statistically significant (p > 0.05). Difference between the two groups in terms of timing, type of delivery and newborn birthweight was not observed. Neither ectopic pregnancy nor multiple pregnancies occurred. "Hour 1" uterine transfer of ICSI oocytes, still to be considered with caution because of the relatively low number of observations of the pilot study, deserves further attention on a larger scale, and might be evaluated for clinical and financial effectiveness in other clinical settings.

Term pregnancy and live birth subsequent to immediate uterine transfer of sperm microinjected oocyte in a natural cycle.

It has been reported that it is possible to achieve a pregnancy after immediate uterine transfer of oocyte and sperm, before fertilization and cleavage were known to have occurred; there is an enormous amount of work about the optimal timing of embryo transfer, with no conclusive evidence of a gold standard satisfying patient age, endometrial receptivity, hormonal levels and embryological parameters. We hereby report a case of one 35-year-old nulligravid woman with longstanding tubal factor infertility and 3 previous failed ICSI cycles, treated with ICSI and immediate transfer of the oocyte microinjected with a spermatozoon in a natural cycle. A single oocyte was retrieved, injected with a spermatozoon and transferred 40 min after injection, resulting in an uneventful pregnancy and delivery of a healthy female infant weighing 3320 g at 40 weeks' estimated gestational age. This case, certainly novel, should be interpreted with caution. Whether confirmed for efficacy and safeness in appropriate controlled clinical trials, our present observation could offer a simple, practical and cost-effective approach in ART programs in selected patients.

Clinical complications after transvaginal oocyte retrieval in 7,098 IVF cycles.

We report the complications observed after transvaginal oocyte retrieval guided by ultrasound in 7,098 IVF cycles. The frequency of severe complications in our patients was 0.08%, of which four cases were intraperitoneal bleeding (0.06%) and two were cases of ovarian abscess (0.003%).

The possible role of hyperhomocysteinemia on IVF outcome.

PURPOSE: To evaluate the effect of the hyperhomocysteinemia on pregnancy rate, implantation rate and abortion rate after IVF. METHOD: Data from a total of 48 infertile couples with hyperhomocysteinemia were prospectively collected for this study. All patients underwent a standard down regulation protocol for ovarian stimulation. Oocytes recovery was performed at 36 h after hCG administration. Embryo transfer took place at 48 h after insemination. The patients were matched in two groups that received or did not receive therapy (group A and B respectively) to normalize homocysteine plasma level. RESULTS: Pregnancy rate, implantation rate and abortion rate varied significantly (p

The impact of the depth of embryo replacement on IVF outcome.

PURPOSE: To evaluate the effect of the depth of embryo transfer replacement on clinical pregnancy rate. METHODS: Data from a total of 104 consecutive embryo transfers performed on 104 women aged 26-37 years were prospectively collected for this study. All patients underwent a standard down regulation protocol for ovarian stimulation. Oocytes retrieval were performed at 36 h after hCG administration. Embryo transfer took place at 48 h after insemination. The patients were matched in two groups according to the distance between the tip of the catheter and the uterine fundus at transfer (group A > 10 < 15 mm and group B < or = 10 mm). The same method of loading embryos into the embryo transfer catheter was used. RESULTS: Clinical pregnancy rates varied significantly (p < or = 0.05) between the two groups: 27.7% in group A and 14% in group B. The number and quality of embryos transferred did not differ between the groups. CONCLUSIONS: The results suggest that the depth of embryo replacement may be an important variable in embryo transfer technique.

Three-dimensional structure of the zona pellucida at ovulation.

The mammalian zona pellucida (ZP) is an extracellular matrix surrounding oocytes and early embryos, which is critical for normal fertilization and preimplantation development. It is made up of three/four glycoproteins arranged in a delicate filamentous matrix. Scanning electron microscopy (SEM) studies have shown that ZP has a porous, net-like structure and/or nearly smooth and compact aspect. In this study, the fine 3-D structure of the human and mouse ZP is reviewed with the aim to integrate ultrastructural and molecular data, considering that the mouse is still used as a good model for human fertilization. By conventional SEM observations, numerous evidences support that the spongy ZP appearance well correlates with mature oocytes. When observed through more sophisticated techniques at high resolution SEM, ZP showed a delicate meshwork of thin interconnected filaments, in a regular alternating pattern of wide and tight meshes. In mature oocytes, the wide meshes correspond to "pores" of the "spongy" ZP, whereas the tight meshes correspond to the compact parts of the ZP surrounding the pores. In conclusion, the traditional "spongy" or "compact" appearance of the ZP at conventional SEM appears to be only the consequence of a prevalence of different arrangements of microfilament networks, according to the maturation stage of the oocyte, and in agreement with the modern supramolecular model of the ZP at the basis of egg-sperm recognition. Despite great differences in molecular characterization of ZP glycoproteins between human and mouse ZP, there are no differences in the 3-D organization of glycoproteic microfilaments in these species.

Urinary follicle-stimulating hormone (FSH) is more effective than recombinant FSH in older women in a controlled randomized study.

OBJECTIVE: The following study was conducted to determine which FSH, recombinant or urinary, works better in older women. DESIGN: We conducted a controlled randomized study in a single university IVF center. SETTING: University IVF center. PATIENT(S): Women (N = 257) over 39 years old undergoing IVF. INTERVENTION(S): The patients were randomized into two study groups at their first IVF cycle: 121 patients were treated with recombinant FSH, and 120 patients were treated with urinary FSH. Both groups were suppressed with a long GnRH analog protocol. MAIN OUTCOME MEASURE(S): Days of stimulation, E2 at the day of hCG, total amount of FSH administered, number of oocytes collected, amount of FSH per oocyte, and number of embryos obtained. RESULT(S): Patients treated with urinary FSH required a significantly lower total amount of FSH, and a lower amount of FSH per oocyte than women treated with recombinant FSH. The other measures evaluated did not show any statistically significant differences. CONCLUSION(S): Our study showed that urinary FSH performed better in older women than recombinant FSH when associated with the long protocol.