Elevated Anti-Müllerian Hormone as a Prognostic Factor for Poor Outcomes of In Vitro Fertilization in Women with Polycystic Ovary Syndrome.

Women with polycystic ovary syndrome (PCOS) tend to have elevated anti-Müllerian hormone (AMH) levels, which appear to correlate with disease severity and pregnancy outcomes. This was a retrospective observational study designed to assess the relationship between circulating AMH levels and in vitro fertilization (IVF) outcomes. The study involved 150 women with PCOS who underwent IVF treatments. The women's IVF cycles were allocated into three subgroups according to AMH levels: 'low' (AMH < 3.7 ng/mL; n = 49), 'middle' (AMH 3.7-7.4 ng/mL; n = 94), and 'high' (AMH > 7.4 ng/mL; n = 56). All pregnancy-related outcomes (positive beta human chorionic gonadotropin (ßHCG), clinical pregnancy rate, live birth rate, and cumulative live birth rate) were greater in women's IVF cycles with 'low' AMH when compared to those with 'middle' or 'high' AMH (p < 0.05). AMH levels below 3.7 ng/mL were found to be associated with lower oocyte immaturity rate and better pregnancy outcomes, although baseline AMH was not shown to have any significant predictive power for live birth and cumulative live birth in the multivariable logistic regression analysis after adjusting for possible confounders nor in the ROC analyses. In summary, the current study lays the groundwork to validate high AMH levels as a poor prognostic factor for pregnancy outcomes after IVF in women with PCOS.

Female Reproductive Health - study of an egg donor population.

OBJECTIVE: Infertility is one of the most significant reproductive health issues addressed with medically assisted procreation. This study looked into a potential correlation between the number of mature oocytes harvested and donor biological characteristics in order to propose an anti-Müllerian hormone (AMH) cutoff level to optimize the selection of candidates for gamete donation. METHODS: The donors were healthy women included in the Public Gamete Bank between 2011 and 2021. Their results can be used as a national indicator of fertility. RESULTS: We found that women with higher AMH levels had more antral follicles and oocytes harvested. As age increased, the number of oocytes harvested decreased. The suggested AMH cutoff level for successful donation was 1.12 ng/mL. CONCLUSIONS: The analysis of the reproductive health of Public Gamete Bank donors allows the standardization of AMH cutoff values at a national level, since the same laboratory techniques were employed consistently across medical centers. The study also allowed insight into the factors that compromise donation success. If adopted, a more rigorous selection of donor candidates would increase the success rate of egg donations.

Preservation of fertility in female patients with hematologic diseases.

Recent developments of assisted reproduction techniques turned possible to avoid the infertility consequences of oncologic treatments, but fertility preservation (FP) has been somewhat neglected in women with hematologic diseases undergoing gonadotoxic treatments. For these specific cases, the current options for FP include the cryopreservation of embryos, mature oocytes and ovarian tissue, and oocyte in-vitro maturation. We intend to make patients and clinicians aware of this important and relevant issue, and provide hematologists, assisted reproduction physicians and patients, with updated tools to guide decisions for FP. The physicians of the units responsible for female FP should always be available to decide on the best-individualized FP option in strict collaboration with hematologists. With a wide range of options for FP tailored to each case, a greater level of training and information is needed among clinicians, so that patients proposed to gonadotoxic treatments can be previously advised for FP techniques in hematological conditions. ABBREVIATED ABSTRACT: Recent developments of assisted reproduction techniques turned possible to preserve the fertility of women with hematologic diseases undergoing gonadotoxic treatments. Current options for fertility preservation in women with hematologic diseases are presented. It is imperative to offer fertility preservation to all women before starting any gonadotoxic treatment and in some cases after treatment. Fertility preservation methods enable to later achieve the desired pregnancy.

Candidates selection for oocyte donation in a public gamete bank-Predictive value of the anti-Müllerian hormone.

Background: Infertility treatments with oocyte donation are becoming frequent. Recruitment of oocyte donors is a demanding and costly process and therefore of crucial importance. The selection of the oocyte donors undergoes a rigorous evaluation process of the candidates with routine measurement of the anti-Müllerian hormone (AMH) levels (ovarian reserve test). Our aim was to assess whether AMH levels could act as a good marker as tool to select the donor candidates and correlate them with the ovarian response to stimulation with a gonadotropin-releasing hormone antagonist protocol as well as to identify and validate the appropriate AMH level threshold by correlating it with the number of oocytes retrieved. Methods: A retrospective analysis of the oocyte donors' clinical records was performed. Results: The mean age of the participants was 27 years. The ovarian reserve evaluation showed a mean AMH of 5.20 ng/mL. An average number of 16 oocytes was retrieved (12 mature oocytes MII). AMH levels showed a statistically significant positive correlation with the number of total oocytes retrieved. A threshold value of AMH = 3.2 ng/mL predictive of the retrieval <12 oocytes (areas under the curve, 0.7364; 95% confidence interval: 0.529-0.944) was identified by receiver operating characteristic curve. Using this cutoff, the normal response (12 oocytes) was predicted with a sensitivity of 77% and a specificity of 60%. Conclusions: The measurement of AMH may be a determining factor in the choice of the oocyte donor candidates to maximize the response to requests from beneficiaries who require donor oocytes to perform assisted reproductive technique cycles.

All that glitters is not gold: a stereological study of human donor oocytes.

Here we report a quantitative analysis of human metaphase II (MII) oocytes from a 22-year-old oocyte donor, retrieved after ovarian-controlled hyperstimulation. Five surplus donor oocytes were processed for transmission electron microscopy (TEM), and a stereological analysis was used to quantify the distribution of organelles, using the point-counting technique with an adequate stereological grid. Comparisons between means of the relative volumes (Vv) occupied by organelles in the three oocyte regions, cortex (C), subcortex (SC) and inner cytoplasm (IC), followed the Kruskal-Wallis test and Mann-Whitney U-test with Bonferroni correction. Life cell imaging and TEM analysis confirmed donor oocyte nuclear maturity. Results showed that the most abundant organelles were smooth endoplasmic reticulum (SER) elements (26.8%) and mitochondria (5.49%). Significant differences between oocyte regions were found for lysosomes (P = 0.003), cortical vesicles (P = 0.002) and large SER vesicles (P = 0.009). These results were quantitatively compared with previous results using prophase I (GV) and metaphase I (MI) immature oocytes. In donor MII oocytes there was a normal presence of cortical vesicles, SER tubules, SER small, medium and large vesicles, lysosomes and mitochondria. However, donor MII oocytes displayed signs of cytoplasmic immaturity, namely the presence of dictyosomes, present in GV oocytes and rare in MI oocytes, of SER very large vesicles, characteristic of GV oocytes, and the rarity of SER tubular aggregates. Results therefore indicate that the criterion of nuclear maturity used for donor oocyte selection does not always correspond to cytoplasmic maturity, which can partially explain implantation failures with the use of donor oocytes.

Genetic counseling and carrier screening in candidates for gamete donation at a Portuguese center.

OBJECTIVE: Genetic counseling and carrier screening are part of the gamete donation process by healthy individuals. We aim to review the findings of genetic counseling and carrier screening of a cohort of candidates at our public gametes bank. METHODS: Thirty-four male and 64 female candidates had genetic counseling with a medical geneticist before donation. Of these, one female candidate voluntarily dropped-out. Thirty-four males and 63 females performed karyotype and screening for the more common pathogenic variants for CFTR-related cystic fibrosis and spinal muscular atrophy (SMN1) in the Portuguese population. In addition, all females also performed Fragile X expansion screening (FMR1). Thirty candidates with known or assumed African ancestry performed hemoglobinopathies screening. RESULTS: Six candidates were definitely or temporarily withheld from the donation process given their family or personal history that required further investigation. Of 97 candidates tested, 16.5% presented anomalous laboratory results (16/97): ten candidates were carriers for an autosomal recessive disorder - cystic fibrosis (5/97), sickle cell anemia (3/30), and spinal muscular atrophy (2/97). One female was an FMR1 pre-mutation carrier (1/63). One female candidate presented with triple X mosaicism: 47,XXX[2]/46,XX[50]. Two candidates presented with chromosomal instability of unknown origin. In one candidate, a mosaic for the Philadelphia chromosome was detected, revealing the diagnosis of chronic myeloid leukemia. CONCLUSIONS: From a cohort of 97 candidates, 21.7% had a family/personal history or an anomalous laboratory result that required additional genetic counseling, stressing the importance of performing pre-donation genetic counseling in this population.

Male Reproductive Health - study of a sperm donor population.

OBJECTIVE: Every individual has the right to a safe and satisfactory sex and reproductive life. Therefore, several countries have made efforts to make Reproductive Health resources available to their populations. However, few results have been published regarding the policies implemented in Portugal. This study looked into the reproductive health status of the Portuguese male population. METHODS: Sperm donor candidates from the Portuguese Public Gamete Bank registered between 2011 and 2018 were included in the study. Spermogram findings were evaluated with respect to sociodemographic and risk factors. RESULTS: This is the first study performed in this population. We found that sperm quality has decreased throughout the last nine years, and that spermatozoa progressive motility is inversely correlated with the body mass index. An association between drug use and decreased sperm pH was also found. CONCLUSIONS: Changes in sperm quality have important consequences in male fertility. Most of the identified causes of decreased sperm quality are modifiable factors and should therefore be addressed since an early age.

Influence of Women´s Residence Region on Assisted Reproduction Treatments - Experience of a Tertiary Center in Northern Portugal.

OBJECTIVE: Data on the possible influence of women´s region of residence, within the same country, on the outcomes of medically assisted reproduction cycles are scarce. This study aims to assess the impact of the women's region of residence on the results of in-vitro fertilization cycles. METHODS: We evaluated in-vitro fertilization cycles between 2010 and 2017, performed in a northern Portugal assisted reproduction center. We defined two groups: Douro Litoral (group 1; n=783), and Trás-os-Montes and Alto Douro (group 2; n=178). We analyzed demographics and cycle-related variables, and we calculated the rates for embryo transfer cycles. We used the Mann-Whitney and Chi-square tests and p<0.05 was considered statistically significant. RESULTS: We included 961 cycles. The region of residence had no effect on the following variables: women´s age; body mass index; or duration of infertility (p>0.05). Group 2 had a statistical significant lower number of previous cycles than group 1 (1.3±0.5 and 1.5±0.7; p=0.005). In the sub-analysis of cycles with embryo transfer (n=781), group 1 obtained had rates of normal fertilization (62.5% vs. 57.5%; p=0.04), miscarriage rate (30.0 vs. 10.9%; p=0.007), and lower implantation rates compared to group 2 (33.3% vs. 50.0%; p<0.001). CONCLUSIONS: Women from the region of Trás-os-Montes e Alto Douro had a lower number of previous cycles, compared to those from the Douro Litoral, despite the absence of statistical significant differences in terms of age or infertility duration. These findings reinforce the need to contemplate the sociodemographic and socioeconomic variables in this context.

Stereological study of organelle distribution in human oocytes at metaphase I.

We have previously presented a stereological analysis of organelle distribution in human prophase I oocytes. In the present study, using a similar stereological approach, we quantified the distribution of organelles in human metaphase I (MI) oocytes also retrieved after ovarian stimulation. Five MI oocytes were processed for transmission electron microscopy and a classical manual stereological technique based on point-counting with an adequate stereological grid was used. Kruskal-Wallis and Mann-Whitney U-tests with Bonferroni correction were used to compare the means of relative volumes (Vv) occupied by organelles. In all oocyte regions, the most abundant organelles were mitochondria and smooth endoplasmic reticulum (SER) elements. No significant differences were observed in Vv of mitochondria, dictyosomes, lysosomes, or SER small and medium vesicles, tubular aggregates and tubules. Significant differences were observed in other organelle distributions: cortical vesicles presented a higher Vv (P = 0.004) in the cortex than in the subcortex (0.96% vs 0.1%) or inner cytoplasm (0.96% vs 0.1%), vesicles with dense granular contents had a higher Vv (P = 0.005) in the cortex than in the subcortex (0.1% vs 0%), and SER large vesicles exhibited a higher Vv (P = 0.011) in the inner cytoplasm than in the subcortex (0.2% vs 0%). Future stereological analysis of metaphase II oocytes and a combined quantitative data of mature and immature oocytes, will enable a better understanding of oocyte organelle distribution during in vivo maturation. Combined with molecular approaches, this may help improve stimulation protocols and in vitro maturation methods.

Structural and molecular analysis of the cancer prostate cell line PC3: Oocyte zona pellucida glycoproteins.

The human oocyte zona pellucida (ZP) is made of four glycoproteins, ZP1-ZP4. Recently, the prostate adenocarcinoma and prostate cancer PC3 cell-line were shown to express the human oocyte ZP3 glycoprotein, which was evaluated in a single report subject to patent. To further clarify whether oocyte zona pellucida glycoproteins are expressed in prostate cancer tissue and PC3-cells, in this report we evaluated protein expression of the four ZP glycoproteins in normal prostate tissue, prostate adenocarcinoma tissue and PC3-cells, and performed quantitative mRNA expression of the four ZP glycoproteins in the PC3 cell-line. Furthermore, as PC3-cells have not yet been studied in detail regarding their ultrastructural characteristics, in the present report we bring forward the detailed ultrastructure of PC3-cells. PC3-cells were divided into pavement and aggregated cells. We observed new ultrastructural features in pavement and aggregated cells, with the later exhibiting two different cell types. In prostate carcinoma tissue and PC3-cells we found protein expression of the four oocyte glycoproteins, ZP1, ZP2, ZP3 and ZP4. In addition, mRNA expression studies revealed expression of ZP1, ZP3 and ZP4 glycoproteins, but not of ZP2. Interestingly, the ZP1 mRNA product exhibited intron retention.

[Successful pregnancies outcomes with the use of in vitro fertilization after Essure® unilateral hydrossalpinx occlusion]. / Gravidezes bem sucedidas com recurso a fertilização in vitro após oclusão de hidrossalpinge unilateral com Essure®

INTRODUCTION: Since two decades we witnessed the publication of several studies devoted to the study of the influence of the presence of hydrosalpinx on the results of embryo transfer techniques. The aim of this study is to present the results of treatment and pregnancy outcomes in women with a history of infertility associated with unilateral hydrosalpinx, visible on vaginal ultrasound, which were subjected to unilateral occlusion with Essure® and subsequent treatment with in vitro fertilization. MATERIAL AND METHODS: We performed a prospective analysis of a sample of 6 women, with a history of infertility and unilateral hydrosalpinx, between April 2010 to May 2013. In all cases we proceeded to unilateral hysteroscopic placement of the Essure® microinsert, prior to performing a cycle for in vitro fertilization. RESULTS: Of the 6 patients undergoing in vitro fertilization, 4 became pregnant (66.7%). Of these, two were uneventful pregnancies until delivery and 2 are still under surveillance, without complications. Of the patients who did not become pregnant after in vitro fertilization (n = 2, 33.3%), 1 conceived spontaneously during the subsequent monitoring. DISCUSSION: The advancement of hysteroscopy in the treatment of hydrosalpinx using the Essure® microinsert placement is a valid alternative to the laparoscopic approach. CONCLUSION: This study suggests the effectiveness of unilateral tubal occlusion caused by Essure® microinsert in improving outcomes of in vitro fertilization treatment in cases of infertility associated with unilateral hydrosalpinx, visible in the vaginal ultrasound.

Introdução: Desde há duas décadas que assistimos à publicação de vários estudos dedicados à avaliação da influência da hidrossalpinge nos resultados de técnicas de transferência de embriões. O objectivo deste trabalho é apresentar os resultados do tratamento e vigilância gestacional em mulheres com história de infertilidade - associada a hidrossalpinge unilateral, visível no exame ecográfico transvaginal - submetidas a oclusão unilateral com Essure® e tratamento subsequente com fertilização in vitro. Material e Métodos: Análise prospectiva de uma amostra constituída por seis mulheres, com antecedentes de infertilidade e hidrossalpinge unilateral, entre Abril de 2010 e Maio de 2013. Em todos os casos procedemos a exclusão de hidrossalpinge recorrendo à colocação unilateral de um dispositivo Essure® por via histeroscópica antes da realização de um ciclo para fertilização in vitro. Resultados: Das seis doentes submetidas a fertilização in vitro, quatro engravidaram (66,7%). Destas, registaram-se duas gravidezes de termo sem intercorrências até ao parto e duas gravidezes encontram-se em vigilância, sem intercorrências conhecidas até à data. Das doentes que não engravidaram após fertilização in vitro (n = 2, 33,3%), uma engravidou espontaneamente durante a vigilância subsequente. Discussão: O avanço da histeroscopia no tratamento de hidrossalpinge, recorrendo à implantação do dispositivo Essure®, representa uma alternativa válida à abordagem laparoscópica. Conclusão: Este estudo sugere que a oclusão tubar unilateral, induzida pelo dispositivo Essure®, melhora os resultados do tratamento com fertilização in vitro em casos de infertilidade associada a hidrossalpinge unilateral, visível no exame ecográfico transvaginal.

Pregnancy after hydrosalpinx treatment with Essure.

We present a case of a 29-year-old woman, with a history of ectopic pregnancy, which required left salpingectomy, and with a tortuous and impermeable right fallopian tube, compatible with hydrosalpinx. As hydrosalpinx itself can compromise a future pregnancy, treatment with Essure was proposed before passing to medically assisted procreation techniques. Five months after placement of Essure in the right fallopian tube, an in vitro fertilisation cycle was successfully completed and the woman had a singleton pregnancy and vaginal delivery without intercurrences.