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1.
Am J Hum Genet ; 108(12): 2238-2247, 2021 12 02.
Artigo em Inglês | MEDLINE | ID: mdl-34798051

RESUMO

Chromosome imbalance (aneuploidy) is the major cause of pregnancy loss and congenital disorders in humans. Analyses of small biopsies from human embryos suggest that aneuploidy commonly originates during early divisions, resulting in mosaicism. However, the developmental potential of mosaic embryos remains unclear. We followed the distribution of aneuploid chromosomes across 73 unselected preimplantation embryos and 365 biopsies, sampled from four multifocal trophectoderm (TE) samples and the inner cell mass (ICM). When mosaicism impacted fewer than 50% of cells in one TE biopsy (low-medium mosaicism), only 1% of aneuploidies affected other portions of the embryo. A double-blinded prospective non-selection trial (NCT03673592) showed equivalent live-birth rates and miscarriage rates across 484 euploid, 282 low-grade mosaic, and 131 medium-grade mosaic embryos. No instances of mosaicism or uniparental disomy were detected in the ensuing pregnancies or newborns, and obstetrical and neonatal outcomes were similar between the study groups. Thus, low-medium mosaicism in the trophectoderm mostly arises after TE and ICM differentiation, and such embryos have equivalent developmental potential as fully euploid ones.


Assuntos
Aneuploidia , Blastocisto , Desenvolvimento Embrionário/genética , Fertilização in vitro , Testes Genéticos , Mosaicismo/embriologia , Blastocisto/patologia , Método Duplo-Cego , Transferência Embrionária , Feminino , Fertilização in vitro/métodos , Humanos , Incidência , Recém-Nascido , Masculino , Gravidez , Resultado da Gravidez , Estudos Prospectivos
2.
J Assist Reprod Genet ; 40(12): 2895-2902, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-37819552

RESUMO

PURPOSE: To assess whether embryo transfer (ET) technique can influence the clinical pregnancy rate (CPR) and its correlation with the embryo transfer difficulty. DESIGN: This single center retrospective cohort analysis of fresh and frozen single blastocyst transfers performed between January 2016 and December 2021 included fresh and frozen single blastocyst transfers performed during the study timeframe. Direct technique was the only one used from January 2016 to September 2017. From September 2017 to March 2019, the choice between the two techniques was given by randomization, due to a clinical trial recruitment. From April 2019, only the afterload technique was used. Preimplantation genetic testing cycles and gamete donation procedures and cycles performed with external gametes or embryos were excluded. CPR was the primary outcome, while difficult transfer rate the secondary one. Univariate and multivariate logistic regressions were performed. RESULTS: During the period, 8,189 transfers were performed. CPR of the afterload group resulted significantly higher compared to the direct group (44.69% versus 41.65%, OR 1.13, 95% CI 1.02-1.25, p = 0.017) and the rate of difficult transfers two-thirds lower (9.06% versus 26.85%, OR 0.27, 95% CI 0.24-0.31, p < 0.001). CONCLUSION: Our study demonstrated that CPR is significantly affected by the ET technique. In particular, with the afterload protocol, both CPR and easy transfer rates increased. TRIAL REGISTRATION: http://clinicaltrials.gov registration number: NCT05364528, retrospectively registered on 3rd of May 2022.


Assuntos
Criopreservação , Transferência Embrionária , Feminino , Humanos , Gravidez , Blastocisto , Estudos de Coortes , Transferência Embrionária/métodos , Fertilização in vitro , Taxa de Gravidez , Estudos Retrospectivos , Ensaios Clínicos Controlados Aleatórios como Assunto
3.
Reprod Biomed Online ; 45(4): 661-668, 2022 10.
Artigo em Inglês | MEDLINE | ID: mdl-35907685

RESUMO

RESEARCH QUESTION: Does the embryologist performing the embryo transfer impact the cycle outcome, in terms of ongoing pregnancy rate (OPR)? DESIGN: This single-centre retrospective study analysed the results, corrected for main confounders, from 28 embryologists and 32 physicians who performed respectively 24,992 and 24,669 fresh embryo transfers (either at cleavage or blastocyst stage) during a 20-year period from January 2000 to December 2019, in a university-affiliated tertiary care assisted reproductive technology (ART) centre. Primary outcome was OPR, defined as the number of viable pregnancies that had completed at least 12 weeks of gestation on the total number of embryo transfers performed. The study also assessed whether the embryologist's experience, measured in terms of number of embryo transfers performed prior to the day of the procedure, had an impact on their performance. The secondary aim was to assess which variable, between the embryologist and physician, more significantly impacted OPR. RESULTS: The overall unadjusted OPR was 22.54%. The embryologist performing the embryo transfer was found to significantly affect the OPR (P < 0.0001), corrected for potential confounders. However, the physician factor made a slightly greater contribution to the model (likelihood ratio 21.86, P < 0.001 versus likelihood ratio 17.20, P < 0.0001). No significant association was found between the experience of the embryologist and OPR (P = 0.067). CONCLUSIONS: These results show how the 'human factor' influences the chances of a positive outcome in the final step of a high-tech procedure and underline the importance of implementing an operator quality performance programme (both for physicians and embryologists) to ensure the maintenance of benchmark results and eventually retrain underperforming operators.


Assuntos
Blastocisto , Transferência Embrionária , Transferência Embrionária/métodos , Feminino , Humanos , Gravidez , Taxa de Gravidez , Técnicas de Reprodução Assistida , Estudos Retrospectivos
4.
J Assist Reprod Genet ; 37(4): 875-882, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31981037

RESUMO

OBJECTIVE: To assess rates of successful testicular sperm retrieval and intracytoplasmic sperm injection (ICSI) outcome in cancer survivors affected by non-obstructive azoospermia (NOA) or retrograde ejaculation (RE)/failure of emission (FOE). METHODS: A retrospective analysis of cancer survivors who did not cryopreserve sperm prior to treatment undergoing testicular sperm extraction (TESE). Non-cancer NOA patients and neurologic RE/FOE were the control group. RESULTS: A total of 97 cancer survivors were offered TESE and 88 (91%) accepted. Sperm was retrieved and cryopreserved in 34/67 patients with NOA (50.7%) and in 21/21 patients affected by RE/FOE (100%). Sperm retrieval rates were similar in the control group (44.9% in NOA and 100% in RE/FOE). The ICSI cumulative pregnancy rate (60%) and live birth rate (40%) per couple in 30 NOA men did not differ from controls (50.0 and 46.5%, respectively; p = 0.399/0.670). The cumulative pregnancy rate (66.7%) and live birth rate (55.6%) in 18 RE/FOE men did not differ from the control group (38.9 and 33.3%, respectively; p = 0.181/0.315). The cancer type and the resulting infertility disorder (NOA or RE/FOE) were not associated with ICSI outcomes. Female partner age was inversely related to the cumulative live birth rate, being fourfold lower (11.5%) in women ≥ 40 years and 48.8% in younger women (p = 0.0037). CONCLUSIONS: The rate of successful TESE and the ICSI outcome in cancer survivors with NOA and RE/FOE is the same as non-cancer azoospermic patients. Female partner age (older than 40 years) was associated with a significant reduction in live birth rates after TESE-ICSI procedures.


Assuntos
Azoospermia/prevenção & controle , Sobreviventes de Câncer , Recuperação Espermática/normas , Espermatozoides , Adulto , Azoospermia/epidemiologia , Azoospermia/patologia , Criopreservação , Feminino , Fertilização in vitro , Humanos , Nascido Vivo , Masculino , Neoplasias/complicações , Neoplasias/patologia , Gravidez , Taxa de Gravidez , Injeções de Esperma Intracitoplásmicas
5.
J Assist Reprod Genet ; 37(5): 1195-1201, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32215826

RESUMO

PURPOSE: To report our experience on homologous intrauterine insemination (IUI) with gonadotropin controlled ovarian stimulation (COS) cycles and to examine different variables which could predict IUI success. MATERIALS AND METHODS: This is a retrospective analysis of IUIs performed between January 1997 and December 2017. A total of 7359 COS IUI's procedures (2901 couples) were reviewed. Clinical pregnancy, live birth rate and age, body mass index (BMI), smoking habit, duration of infertility, sperm characteristics before and after treatment (total motile count, morphology, and vitality), day 3 FSH, total gonadotropin dose, and number of follicles were assessed by multivariate logistic regression analysis, and data were expressed as odds ratio (OR). RESULTS: The mean female age at the time of COS was 35.10 ± 3.93 years. The most common single infertility diagnoses were unexplained infertility (53.55%), mild male factor (19.69%), and anovulation (10.95%). The total progressive motile sperm count (TPMC) was > 1 × 106/ml (mean 1.34 ± 1.08 × 106/ml). The clinical pregnancy rate was 9.38%, and the live birth rate was 7.19% per cycle. Twin pregnancies were 12.17%. Cumulative pregnancy was 21.89% and cumulative live birth rate was 17.58% per couple. Clinical pregnancy and live birth rates were significantly associated with female age [OR 0.97 (95% CI 0.95-0.99) and 0.95 (95% CI 0.93-0.97), respectively] and day 3 FSH [OR 0.91 (95% CI 0.87-0.94) e 0.90 (95% CI 0.87-0.94), respectively]. CONCLUSIONS: Clinical pregnancy rate and live birth rates after COS-IUIs were significantly influenced by female age and FSH levels. TRIAL REGISTRATION: Clinical trial registration number: NCT03836118.


Assuntos
Fertilização in vitro , Infertilidade Feminina/terapia , Idade Materna , Indução da Ovulação , Adulto , Feminino , Gonadotropinas/uso terapêutico , Humanos , Infertilidade Feminina/fisiopatologia , Inseminação Artificial , Nascido Vivo/epidemiologia , Masculino , Folículo Ovariano/crescimento & desenvolvimento , Folículo Ovariano/metabolismo , Folículo Ovariano/patologia , Gravidez , Taxa de Gravidez , Prognóstico , Espermatozoides/crescimento & desenvolvimento , Espermatozoides/patologia
6.
Minerva Pediatr ; 72(3): 182-195, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32274912

RESUMO

The first 1000 days of life represent a critical window for infants' and children's development. Overweight and insulin resistance, at the basis of non-communicable diseases (NCDs), are linked to various risk factors that begin in childhood, including children's diet. Italian data on infants' and children's dietary habits show higher intake of proteins, simple sugars, unhealthy fats and salt than recommended, while the iron intake is below requirement. We reviewed current literature analyzing observational studies, meta-analysis, systematic review and randomized clinical trials of the last 10 years (from 2009) on nutrition in developmental age, providing some few rules to abide by. Exclusive breastfeeding is recommended by World Health Organization for the first 6 months of life and it should be continued alongside the complementary feeding period until 12 months, or even afterward. Complementary feeding should not be started before the 17th week of age with energetically adequate foods, paying attention to limit protein intake and favoring iron-rich foods. Intake of simple sugars should be limited or avoided at all; it has been demonstrated that substituting sugar-sweetened beverages with water decreases body fatness development in adolescence. Quality of the ingested fats is more important than their quantity: polyunsaturated fatty acids should be preferred. Sodium intake should be limited in the first 24 months of life, as first prevention measure of arterial hypertension later in adulthood. Healthy eating habits are the first important step toward the prevention of NCDs.


Assuntos
Comportamento Alimentar/fisiologia , Fenômenos Fisiológicos da Nutrição do Lactente/fisiologia , Estado Nutricional/fisiologia , Aleitamento Materno , Pré-Escolar , Dieta , Humanos , Lactente , Recém-Nascido , Resistência à Insulina , Obesidade Infantil/epidemiologia , Fatores de Risco
7.
J Assist Reprod Genet ; 36(12): 2493-2504, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31713776

RESUMO

PURPOSE: To report the effects of blastocyst stage aneuploidy testing on clinical, gestational, and neonatal outcomes for patients of advanced maternal age undergoing IVF. METHODS: This is a single-center observational-cohort study with 2 years follow-up. The study includes a total of 2538 couples undergoing 2905 egg collections (control group), 308 (PGT-A), and 106 (drop-out group, consenting for PGT-A but withdrawing due to poor embryological outcome) RESULTS: Compared with control group, PGT-A showed improved clinical outcomes (live-birth rate per transferred embryo, LBR 40.3% vs 11.0%) and reduced multiple pregnancy rate (MPR, 0% vs 11.1%) and pregnancy loss (PL, 3.6% vs 22.6%). Drop-out group showed the worst clinical outcomes suggesting that abandoning PGT-A due to poor response to ovarian stimulation is not a favorable option. Cytogenetic analysis of product of conceptions and CVS/amniocentesis showed higher aneuploid pregnancy rates for control group regardless of embryo transfer strategy (0%, 17.9%, and 19.9%, for PGT-A, control day 5 and day 3, respectively). Multivariate analysis showed no negative impact of PGT-A-related interventions on cumulative delivery rate (26.3%, 95% CI 21.5-31.6 vs 24.0%, 95% CI 22.5-25.6 for PGT-A and control, respectively) and on neonatal outcomes. CONCLUSION: PGT-A improves clinical outcomes, particularly by reducing pregnancy loss and chromosomally abnormal pregnancy for patients of advanced maternal age, with no major impact on cumulative live-birth rate (CLBR) per egg retrieval.


Assuntos
Aborto Espontâneo/diagnóstico , Aneuploidia , Idade Materna , Diagnóstico Pré-Implantação , Aborto Espontâneo/epidemiologia , Aborto Espontâneo/genética , Aborto Espontâneo/fisiopatologia , Adulto , Blastocisto/citologia , Hibridização Genômica Comparativa , Transferência Embrionária , Feminino , Fertilização in vitro/métodos , Testes Genéticos , Humanos , Gravidez , Taxa de Gravidez
8.
Hum Reprod ; 33(11): 1992-2001, 2018 11 01.
Artigo em Inglês | MEDLINE | ID: mdl-30265329

RESUMO

STUDY QUESTION: Are trophectoderm biopsy or other pre-vitrification features or laboratory practices associated with differences in blastocyst post-warming behavior (degeneration, re-expansion and live birth after single embryo transfer (SET))? SUMMARY ANSWER: Blastocyst morphology, day of full development and artificial shrinkage (either laser-assisted or biopsy-induced) are the pre-vitrification parameters/practices most strongly associated with post-warming behavior and implantation potential while there was no association with trophectoderm biopsy. WHAT IS KNOWN ALREADY: Since the introduction of vitrification, the adoption of cycle segmentation, freeze-all and SET policies, as well of trophectoderm biopsy-based aneuploidy testing (i.e. pre-implantation genetic testing for aneuploidies (PGT-A)), the number of blastocysts vitrified worldwide has increased greatly. Previous studies already defined generally high blastocyst cryo-survival rates after vitrification-warming (>95%), along with a positive correlation between blastocyst re-expansion and morphology with implantation. Additionally, artificial shrinkage has been outlined as a potentially beneficial procedure, while the association between embryo cryo-survival and trophectoderm biopsy is still unclear. STUDY DESIGN, SIZE, DURATION: Cohort study conducted at two IVF centers (1 and 2). A total of 2129 consecutive SETs using vitrified-warmed blastocysts in either non-PGT or PGT-A cycles between June 2016 and August 2017 were included. A freeze-all strategy was in place and three main pre-vitrification practices were used: (i) no biopsy and no artificial shrinkage (Clinic 1); (ii) trophectoderm biopsy and vitrification of collapsed blastocyst within 30 min (Clinics 1 and 2); and (iii) no biopsy but laser-assisted artificial shrinkage (Clinic 2). The primary outcome was the blastocyst degeneration rate. Overall, 2108 surviving blastocysts were graded at 1.5 h after warming for degeneration (absent or partial) and re-expansion (full, partial or absent) grades and post-warming morphological quality. Logistic regression analyses were conducted to assess the association of any pre-vitrification feature with blastocyst post-warming behavior. The logistic regressions conducted upon live birth after either untested or euploid SET also included the post-warming characteristics. PARTICIPANTS/MATERIALS, SETTING, METHODS: Center 1 is a private IVF facility, while center 2 is the IVF facility of a University hospital. In non-PGT cycles, ICSI with blastocyst culture up to full-expansion and vitrification were performed. At center 1 the untested blastocysts were vitrified when still expanded, while at center 2 they underwent laser-assisted artificial shrinkage. In PGT-A cycles, in both clinics, trophectoderm biopsy (which involves laser-assisted shrinkage) was done without previous zona-opening on Day 3, and vitrification was performed within 30 min whilst the blastocyst remained collapsed. A qPCR-based chromosome analysis was conducted. Only SETs were performed (euploid-SET in case of PGT-A). Any cycle-, laboratory- and embryo-based feature which could impact blastocyst post-warming behavior was included in the analyses as putative confounder. MAIN RESULTS AND THE ROLE OF CHANCE: The overall degeneration rate was 1% (N = 21/2129). The results were consistent among different vitrification/warming operators or kits used, as well as any other IVF laboratory-related parameter. Blastocyst artificial shrinkage (either laser-assisted or biopsy-induced) involved a lower risk of degeneration after warming (odds ratio (OR) [95% CI] = 0.26 [0.09-0.79]). Conversely, both poor morphological quality pre-vitrification and taking 7 days to reach full blastulation resulted in a significantly higher risk (OR [95% CI] = 11.67 [3.42-39.83] and 4.43 [1.10-20.55], respectively). Importantly, trophectoderm biopsy did not show any association with blastocyst cryo-survival/degeneration. Overall 2.5% (N = 53/2108) blastocysts failed to re-expand post-warming. The only parameters significantly associated with no blastocyst re-expansion post-warming were average (OR [95% CI] = 4.96 [2.20-11.21]) or poor (OR [95% CI] = 19.54 [8.39-45.50]) morphological quality and taking 7 days to reach full blastulation (OR [95% CI] = 3.19 [1.23-8.29]), as well as prevention of spontaneous hatching pre-vitrification (OR [95% CI] = 0.10 [0.01-0.85]). The post-warming features of the survived blastocyst (i.e. degeneration and re-expansion scores and morphological quality) showed no significant association with vitrified blastocyst competence (i.e. live birth) when corrected for pre-vitrification ones (i.e. morphological quality, day of full development and, for untested SET, maternal age at oocyte retrieval). Of note, poor-quality blastocysts pre-vitrification showed a high overall cryo-survival rate post-warming 92.8% (N = 116/125), but the live birth rates were only 2.1% (N = 1/48) and 7.3% (N = 5/68) after untested and euploid SET, respectively. LIMITATIONS, REASONS FOR CAUTION: This study is not randomized and the populations of patients undergoing either non-PGT or PGT-A cycles were different. Centers 1 and 2 adopted different pre-vitrification practices for non-biopsied blastocysts, according to their own laboratory policy. To this regard, multivariate logistic regression analyses were conducted for all outcomes under investigation. WIDER IMPLICATIONS OF THE FINDINGS: Pre-vitrification features may be used to assist selection of competent embryos, moreover, these results allay concern that trophectoderm biopsy might be associated with impaired blastocyst quality or competence after vitrification/warming. STUDY FUNDING/COMPETING INTEREST(S): None. TRIAL REGISTRATION NUMBER: None.


Assuntos
Blastocisto , Criopreservação/métodos , Técnicas de Cultura Embrionária/métodos , Implantação do Embrião , Transferência Embrionária/métodos , Vitrificação , Biópsia , Estudos de Coortes , Feminino , Humanos , Nascido Vivo , Gravidez
9.
Hum Reprod ; 33(10): 1839-1846, 2018 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-30239718

RESUMO

STUDY QUESTION: Can a second round of biopsy, vitrification and chromosomal testing provide a valid diagnosis where the first attempt fails? SUMMARY ANSWER: The risk of inconclusive chromosomal-assessment after trophectoderm biopsy was 2.5% but a further biopsy and vitrification-warming appeared not to impair the competence of euploid blastocysts. WHAT IS KNOWN ALREADY: The increasing implementation of multicell trophectoderm biopsy has significantly reduced the risk of inconclusive diagnosis after preimplantation-genetic-testing (PGT). Yet, few reports have defined the variables that influence the risk of failure or described the technical and clinical outcomes after re-biopsy. STUDY DESIGN, SIZE, DURATION: Retrospective multicenter study involving 8990 blastocyst biopsies conducted between April 2013 and September 2017 at six IVF centers but analyzed at a single genetic laboratory. A total of 206 blastocysts were successfully re-biopsied after warming and re-expansion, then re-vitrified. And 49 of these blastocysts were diagnosed euploid and used in single-embryo-transfers (SETs). Logistic regression analyses were conducted. PARTICIPANTS/MATERIALS, SETTING, METHODS: A total of 3244 PGT-for-aneuploidies (PGT-A) cycles with a freeze-all approach, vitrification and qPCR-based analysis were performed by 2687 consenting couples. DNA amplification failure (AF) or non-concurrent data resulted in inconclusive diagnoses. In case of DNA amplification, the cellularity of the biopsy was estimated according to a previously validated method. Euploid SETs were performed. Clinical pregnancy, miscarriage, live birth rates (LBR) and perinatal outcomes were monitored. MAIN RESULTS AND THE ROLE OF CHANCE: Overall, 2.5% of trophectoderm biopsies resulted in an inconclusive diagnosis (N = 228/8990). Specifically, 2% (N = 176/8990) resulted in AF and 0.5% (N = 52/8990) in non-concurrent results. The only parameters significantly associated with inconclusive diagnoses were the IVF center and the embryo age (days) at biopsy. Among samples with successful amplification, the number of cells in the biopsy and the day of biopsy were critical to limit non-concurrent results. In total, 213 blastocysts with an inconclusive diagnosis were warmed for re-analysis and the survival rate was 96.7% (N = 206/213). The euploidy rate in blastocysts biopsied twice was 51.9% (N = 107/206) and the euploid embryos were re-vitrified. Overall, 49 euploid embryos were warmed for replacement and all survived. The LBR after SET was 38.8% (N = 19/49). No minor/major obstetrical/perinatal complication was reported. LIMITATIONS, REASONS FOR CAUTION: A single aneuploidy-testing method was adopted in this retrospective analysis. A more powered report of the clinical and obstetrical/perinatal outcomes after re-biopsied and re-vitrified blastocysts euploid SET requires a larger sample size. WIDER IMPLICATIONS OF THE FINDINGS: It is important to re-biopsy and re-vitrify undiagnosed blastocysts since healthy live births can result from them. STUDY FUNDING/COMPETING INTEREST(S): None. TRIAL REGISTRATION NUMBER: None.


Assuntos
Aneuploidia , Transferência Embrionária/métodos , Doenças Genéticas Inatas/diagnóstico , Diagnóstico Pré-Implantação/métodos , Adulto , Blastocisto , Criopreservação/métodos , Feminino , Testes Genéticos/métodos , Humanos , Infertilidade , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase em Tempo Real , Estudos Retrospectivos , Transferência de Embrião Único
10.
J Assist Reprod Genet ; 35(3): 457-465, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29168022

RESUMO

PURPOSE: Is there a difference in implantation and pregnancy rates between embryos transferred electively at cleavage or blastocyst stage in infertile women ≤ 38 years with at least four zygotes on day 1 post retrieval? METHODS: A randomized clinical trial was conducted in a single tertiary care hospital with a sample size of 194 patients in each arm for a total population of 388 women. Patients less than 39 years of age with more than three fertilized oocytes and less than four previous assisted reproductive technology (ART) attempts were inclusion criteria. RESULTS: The two groups were similar for age, years of infertility, indication to treatment, basal antimüllerian hormone and FSH, number of previous ART cycles, primary or secondary infertility, type of induction protocol, days of stimulation, total gonadotrophin dose, and estradiol (E2) and progesterone (P) levels at trigger. No statistically significant differences were found in terms of number of retrieved oocytes, inseminated oocytes, fertilization rate, canceled transfers (7.73% in blastocyst and 3.61% in cleavage stage group), and cycles with frozen embryos and/or oocytes. Although a higher number of fertilized oocytes were in the blastocyst stage group (6.18 ± 1.46 vs 5.89 ± 1.54, p = 0.052), a statistically greater number of embryos/randomized cycle were transferred at cleavage stage (1.93 ± 0.371) compared with the number of transferred blastocysts (1.80 ± 0.56), probably due to the number of embryos not reaching blastocyst stage (3.09%). The implantation rate (28.37 vs 25.67%), pregnancy rate per cycle (36.06 vs38.66%), transfer (39.66 vs 40.11%), spontaneous abortions (19.72% vs 12.00%), delivery rate per cycle (27.84 vs 32.99%), and transfer (30.17 vs 34.22%) were not significantly different between the blastocyst and cleavage stage groups. The twin delivery rate was higher in the blastocyst stage group, although not significant (42.59 vs 28.12%). The mean numbers of frozen blastocyst (2.30 ± 1.40 vs 2.02 ± 1.00) and frozen oocytes (7.09 ± 3.55vs 6.79 ± 3.26) were not significantly different between the two groups. CONCLUSIONS: Fresh blastocyst-stage transfer versus cleavage-stage transfer did not show any significant difference in terms of implantation and pregnancy rate in this selected group of patients. A high twin delivery rate in both groups (35.59%) was registered, and although not significant, they were higher in the blastocyst transfer group (42.59 vs 28.12%). Our conclusion supports considering single embryo transfer (SET) policy, even in cleavage stage in patients younger than 39 years with at least four zygotes. TRIAL REGISTRATION: ClinicalTrials.gov registration number NCT02639000.


Assuntos
Blastocisto/fisiologia , Transferência Embrionária/métodos , Adulto , Blastocisto/citologia , Implantação do Embrião , Transferência Embrionária/efeitos adversos , Feminino , Fertilização in vitro , Humanos , Recém-Nascido , Infertilidade Feminina , Masculino , Recuperação de Oócitos , Gravidez , Resultado da Gravidez , Taxa de Gravidez , Gravidez Múltipla , Transferência de Embrião Único , Resultado do Tratamento
11.
J Assist Reprod Genet ; 35(11): 2049-2056, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30097765

RESUMO

PURPOSE: To analyse the impact of female characteristics on assisted reproductive technology outcome among male haematological cancer survivors. METHODS: A retrospective analysis of 93 haematological cancer survivors attending our tertiary referral fertility centre between June 1998 and June 2017 for achieving fatherhood with assisted reproductive technology treatments. RESULTS: A progressive increase in the median female age was observed during the study period (32.2 years until the year 2007 and 36.9 years from the year 2012). Fifty-five out of 93 patients were treated with intracytoplasmic sperm injection (ICSI) (113 ovarian stimulations, 108 ICSI procedures). Cryopreserved ejaculated sperm was used in 28 couples, fresh sperm in 19, and thawed testicular sperm in 8 couples. Mean female age at ovarian stimulation was 37.0 ± 4.7 years. Twenty-six pregnancies resulted in a full-term birth (23% per started ovarian stimulation; 43.6% per couple) and 33 children were born. No significant differences were observed according to source of sperm (fresh, frozen, testicular) and multivariate analysis confirmed that maternal age was the only variable inversely related to the cumulative delivery rate, being five times lower (15.7%) when the female partner was ≥ 40 years (OR = 0.22, 95% CI 0.06-0.77) vs. 58.3% with younger women (p = 0.0037). CONCLUSIONS: Delayed childbearing and female ageing affect ICSI outcome in couples where the male is a survivor of haematological cancer. This topic should be discussed when counselling male cancer patients about fertility preservation.


Assuntos
Envelhecimento , Neoplasias Hematológicas/complicações , Infertilidade Masculina/etiologia , Idade Materna , Comportamento Reprodutivo , Técnicas de Reprodução Assistida/efeitos adversos , Sobreviventes , Adulto , Criança , Feminino , Humanos , Masculino , Estudos Retrospectivos
12.
J Assist Reprod Genet ; 34(2): 225-238, 2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-27924460

RESUMO

PURPOSE: The etiology of maternal aging, a common cause of female factor infertility and a rate-limiting step in vitro fertilization (IVF) success, remains still unclear. Proteomic changes responsible for the impaired successful pregnancy outcome after IVF with aged blastocysts have not been yet evaluated. The objective of this prospective study was to employ proteomic techniques and bioinformatic tools to enlight differences at the protein level in blastocoel fluid of aged and younger woman. METHODS: Protein composition of human blastocoel fluid isolated by micromanipulation from 46 blastocysts of women aged <37 years (group A) and 29 of women aged ≥37 years (group B) have been identified by a shotgun proteomic approach based on high-resolution nano-liquid chromatography electrospray-ionization-tandem mass spectrometry (nLC-ESI-MS/MS) using label free for the relative quantification of their expression levels. RESULTS: The proteomic analysis leads to the identification and quantification of 148 proteins; 132 and 116 proteins were identified in groups A and B, respectively. Interestingly, the identified proteins are mainly involved in processes aimed at fine tuning embryo implantation and development. Among the 100 proteins commonly expressed in both groups, 17 proteins are upregulated and 44 downregulated in group B compared to group A. Overall, the analysis identified 33 proteins, which were increased or present only in B while 76 were decreased in B or present only in A. CONCLUSIONS: Data revealed that maternal aging mainly affects blastocyst survival and implantation through unbalancing the equilibrium of the ubiquitin system known to play a crucial role in fine-tuning several aspects required to ensure successful pregnancy outcome.


Assuntos
Transferência Embrionária , Fertilização in vitro , Biossíntese de Proteínas/genética , Proteômica , Adulto , Fatores Etários , Blastocisto/fisiologia , Sobrevivência Celular , Implantação do Embrião/fisiologia , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Humanos , Idade Materna , Gravidez , Resultado da Gravidez , Espectrometria de Massas em Tandem
13.
Reprod Biol Endocrinol ; 14(1): 55, 2016 Sep 06.
Artigo em Inglês | MEDLINE | ID: mdl-27600956

RESUMO

BACKGROUND: Preimplantation genetic screening (PGS) provides an opportunity to eliminate a potential implantation failure due to aneuploidy in infertile couples. Some studies clearly show that twins following single embryo transfer (SET) can be the result of a concurrent natural conception and an incidence as high as 1 in 5 twins has been reported. In our case PGS was performed on trophectoderm (TE) biopsies by quantitative polymerase chain reaction (qPCR). The product of conception (POC) was cytogenetically investigated after selection of the placental villi by means of the direct method. Molecular cytogenetic characterization of the POC was performed by fluorescence in situ hybridization (FISH) and array-comparative genomic hybridization (a-CGH) analyses. To investigate the possibility of a spontaneous conception, a panel of 40 single nucleotide polymorphisms (SNPs) was used to compare genetic similarity between the DNA of the POC and the DNA leftover of the TE biopsy. FINDINGS: We describe a 36-year old infertile woman undergoing PGS who had a spontaneous abortion after a single euploid embryo transfer on a spontaneous cycle. The POC showed a 45,X karyotype confirmed by FISH and a-CGH. DNA fingerprinting demonstrated a genetic similarity of 75 % between the DNA of the POC and TE biopsy, consistent with a sibling status. All supernumerary euploid embryos were also tested showing a non-self relationship with the POC, excluding a mix-up event at the time of fetal embryo transfer. CONCLUSIONS: DNA fingerprinting of the transferred blastocyst and POC, confirmed the occurrence of a spontaneous conception. This case challenges the assumption that a pregnancy after assisted reproductive technology (ART) is always a result of ART, and strengthens the importance to avoid intercourses during PGS and natural transfer cycles. Moreover, cytogenetic analysis of the POCs is strongly recommended along with fingerprinting children born after PGS to see what the concordance is between the embryo transferred and the resultant child.


Assuntos
Aneuploidia , Impressões Digitais de DNA/métodos , Transferência Embrionária/métodos , Fertilização/genética , Testes Genéticos/métodos , Diagnóstico Pré-Implantação/métodos , Aborto Espontâneo/diagnóstico , Aborto Espontâneo/genética , Aborto Espontâneo/terapia , Adulto , Feminino , Humanos , Infertilidade Feminina/diagnóstico , Infertilidade Feminina/genética , Infertilidade Feminina/terapia , Gravidez
14.
Gynecol Endocrinol ; 32(3): 210-2, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26608547

RESUMO

The role of oxidative stress (OS) in female reproduction is an understudied area for investigation. The aim of the study was to assess reactive oxygen species (ROS) levels both in follicular fluid (FF) and in serum samples of women undergoing IVF and to relate them to clinical outcomes. A total of 56 women were enrolled. From each patient a sample of serum and FF were collected at oocyte retrieval day and analyzed for OS, by measuring a class of reactive oxygen metabolites (ROMs) (dROMs test) and for biological anti-oxidant potential levels (BAP test). The data analyzed in serum and in FF were related to pregnancy outcomes. There was no significant relationship between d-ROMs and BAP levels in serum or FF GROUP A and B and the clinical parameters of prognosis of women: age, number of oocytes retrieved number of oocytes methaphase II, FSH, fertilization rate and cleavage rate, number of embryos transferred and embryo score. There was a positive association between systemic levels of OS measured in serum GROUP A and pregnancy outcomes (p = 0.007) suggesting that the evaluation of OS in FF needs more investigation about markers in follicular microenvironment in order to predict IVF success.


Assuntos
Líquido Folicular/química , Estresse Oxidativo , Espécies Reativas de Oxigênio/sangue , Técnicas de Reprodução Assistida , Adulto , Feminino , Humanos , Gravidez
15.
J Assist Reprod Genet ; 33(4): 461-6, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-26781264

RESUMO

PURPOSE: This study aims to compare implantation, pregnancy, and delivery rates in frozen transfer cycles with blastocysts that were vitrified either with artificial shrinking (AS group) or without (NAS group). METHODS: Retrospective comparative study of artificial shrinking of blastocysts prior to vitrification and frozen embryo transfer cycles in infertile patients undergoing frozen embryo transfer (FET) was done at the Humanitas Fertility Center between October 2009 and December 2013. Main outcome measure(s) were implantation (IR), pregnancy (PR), and delivery rates (DR) between the two groups. RESULTS: A total of 1028 consecutive warming blastocyst transfer cycles were considered. In 580 cycles (total of 822 blastocysts), artificial shrinking was performed prior to vitrification (AS group), while in the remaining 448 cycles (total of 625 blastocysts), the artificial shrinking was not performed (NAS group). There were no differences in patient age (36.4 ± 3.7 vs. 36.3 ± 3.9) and number of embryos transferred (1.41 ± 0.49 vs. 1.38 ± 0.50) between groups. The IR, PR, and DR in the AS group were significantly higher (p < 0.05) than in the NAS group (29.9 vs. 23.0 %, 36.3 vs. 27.9 %, and 26.5 vs. 18.1 %, respectively). CONCLUSIONS: Performing AS of blastocysts prior to vitrification appears to improve implantation, pregnancy, and delivery rates probably related to a decreased risk of ultrastructural cryodamages, plausible when cryopreserving expanded blastocysts.


Assuntos
Blastocisto/fisiologia , Técnicas de Cultura Embrionária/métodos , Técnicas de Reprodução Assistida , Vitrificação , Adulto , Criopreservação , Implantação do Embrião/fisiologia , Transferência Embrionária , Feminino , Humanos , Gravidez , Resultado da Gravidez , Taxa de Gravidez
16.
J Assist Reprod Genet ; 33(12): 1559-1570, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-27586998

RESUMO

PURPOSE: Our aim was to evaluate the ultrastructure of human metaphase II oocytes subjected to slow freezing and fixed after thawing at different intervals during post-thaw rehydration. METHODS: Samples were studied by light and transmission electron microscopy. RESULTS: We found that vacuolization was present in all cryopreserved oocytes, reaching a maximum in the intermediate stage of rehydration. Mitochondria-smooth endoplasmic reticulum (M-SER) aggregates decreased following thawing, particularly in the first and intermediate stages of rehydration, whereas mitochondria-vesicle (MV) complexes augmented in the same stages. At the end of rehydration, vacuoles and MV complexes both diminished and M-SER aggregates increased again. Cortical granules (CGs) were scarce in all cryopreserved oocytes, gradually diminishing as rehydration progressed. CONCLUSIONS: This study also shows that such a membrane remodeling is mainly represented by a dynamic process of transition between M-SER aggregates and MV complexes, both able of transforming into each other. Vacuoles and CG membranes may take part in the membrane recycling mechanism.


Assuntos
Membrana Celular/ultraestrutura , Criopreservação , Retículo Endoplasmático Liso/ultraestrutura , Oócitos/ultraestrutura , Feminino , Congelamento/efeitos adversos , Humanos , Metáfase , Microscopia Eletrônica de Transmissão , Mitocôndrias/ultraestrutura , Vacúolos/ultraestrutura
17.
Gynecol Endocrinol ; 30(1): 48-52, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-24147853

RESUMO

OBJECTIVE: To determine whether intracytoplasmic sperm injection (ICSI) outcome with testicular sperm is superior to that of ejaculated sperm in men with incomplete necrozoospermia, defined as sperm viability ≥5 to ≤45%. STUDY DESIGN: Retrospective study at a Reproductive Medicine Center of a tertiary referral hospital. A total of 231 couples with male infertility due to incomplete necrozoospermia underwent 342 ICSI cycles with freshly ejaculated sperm (ICSI-ejaculated) and 182 cycles with testicular sperm (ICSI-TESE). RESULTS: Overall 1624 MII oocytes were injected in the ICSI-ejaculated group with a fertilisation rate of 60.8%, while in ICSI- TESE cycles the fertilisation rate was 59.6% in 874 MII oocytes. The cleavage rate was higher in ICSI-ejaculated cycles than in ICSI-TESE cycles (96.3% versus 92.9%; p = 0.004). However, the pregnancy and implantation rates per cycle were significantly higher in the ICSI-TESE group (67/182, 36.8% versus 68/342, 19.9% (p = 0.0001); and 23.7% versus 12.7% (p = 0.0001), respectively). The miscarriage rate was similar (ICSI-ejaculated 26.5% versus ICSI-TESE 17.9%, p = 0.301). Live birth rate per cycle in the ICSI-ejaculated group was significantly lower than in the ICSI-TESE (13.7% versus 28.6%, p = 0.0001). CONCLUSIONS: In cases of persistent necrozoospermia, testicular sperm should be favoured over ejaculated sperm. These data call for more research to understand the pathophysiology of refractory necrozoospermia.


Assuntos
Infertilidade Masculina/terapia , Injeções de Esperma Intracitoplásmicas , Recuperação Espermática , Espermatozoides/citologia , Adulto , Sobrevivência Celular , Ejaculação , Feminino , Humanos , Masculino , Gravidez , Taxa de Gravidez , Estudos Retrospectivos , Espermatozoides/fisiologia , Resultado do Tratamento , Adulto Jovem
18.
Front Endocrinol (Lausanne) ; 15: 1466675, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39449747

RESUMO

Background: The management of Non-Obstructive (NOA) Azoospermia or Obstructive Azoospermia (OA) patients relies on testicular sperm extraction (TESE) followed by intracytoplasmic sperm injection (ICSI). In NOA patients the sperm recovery is successful in only 50% of cases and therefore the ability to predict those patients with a high probability of achieving a successful sperm retrieval would be a great value in counselling the patient and his partner. Several studies tried to suggest predictors of a positive TESE (e.g. FSH concentration), but most concluded that diagnostic testicular biopsy (histology) is best. Methods: This is a retrospective analysis of 526 TESE patients. After the extraction of the testis, the resulting sample was immediately given to the embryologist, who examined the tubules for sperm cryopreservation. During the same procedure, a different specimen was destined to the histological analysis. The comparison between the two methodological approaches was carried out through a score. Results: Concordance between TESE and testicular histology outcomes was found in 70,7% of patients; discordance was found in 29,3% of patients. Among the discordance outcomes, in approximately 95% we found at least 1 sperm in the TESE retrieval, while the histology report did not find any spermatozoa or found not enough compared to our evaluation; in only 5% of cases we did not find any spermatozoa or found not enough compared to what was detected in the testicular histology. Conclusion: Based on our experience, to increase diagnostic accuracy, a larger biopsy should be sent to the histopathology laboratory; another option may be to use TESE cell suspension (the same embryologists employ for cryopreservation) for cytological evaluation of spermatogenesis.


Assuntos
Azoospermia , Recuperação Espermática , Testículo , Masculino , Humanos , Testículo/patologia , Estudos Retrospectivos , Azoospermia/patologia , Azoospermia/diagnóstico , Adulto , Injeções de Esperma Intracitoplásmicas/métodos , Criopreservação , Biópsia/métodos , Espermatozoides/patologia
19.
Eur J Obstet Gynecol Reprod Biol ; 302: 232-237, 2024 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-39321724

RESUMO

OBJECTIVE: To study if social and emotional, communicative, cognitive, motor development of children born from cryopreserved oocytes are comparable to general population. DESIGN: Survey study. EXPOSURE: This is a single-center population study, focusing on all children born from cycles of in vitro fertilization of cryopreserved oocytes at Humanitas Fertility Center from January 1st,2003, until December 31st, 2021. MAIN OUTCOME MEASURES: General information about birth history were collected from all parents. Then, screening for developmental delay was performed by administration of a questionnaire, edited according to milestones set for age by the Center for Disease Control and Prevention. Primary outcomes included the percentage of children, for each age range, that reached related milestones. In children younger than 6 years of age, milestones were considered to be reached by each age-based population subgroup studied when ≥ 75 % of included children expressed the tested competences. RESULTS: A total of 355 live births fulfilled the inclusion and exclusion criteria and were included in the study. Results showed that each age-based population subgroup, younger than 6 years of age, scored ≥ 75 % in all the milestones included in the questionnaire. In preadolescents and adolescents, it was noted a decrease in unstructured in-person socializing. Indeed, milestones that scored the lowest values, based on parents' judgement, included tasks concerning peer-to-peer interaction (72.31%) and sexuality and sentimental relationships (63.08%). CONCLUSIONS: Results indicated that the achievement of emotional, communicative, cognitive, and motor milestones in infants, toddlers, and preschoolers was comparable to the general pediatric population. Similarly, emotional wellbeing and social engagement in school-aged children, preadolescents, and adolescents were in line with their peers.


Assuntos
Desenvolvimento Infantil , Criopreservação , Humanos , Feminino , Criança , Pré-Escolar , Masculino , Adolescente , Cognição/fisiologia , Lactente , Emoções , Oócitos , Fertilização in vitro/psicologia , Adulto , Comunicação , Destreza Motora/fisiologia
20.
Front Endocrinol (Lausanne) ; 14: 1150951, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37867514

RESUMO

To date, infertility affects 10% to 15% of couples worldwide. A male factor is estimated to account for up to 50% of cases. Oral supplementation with antioxidants could be helpful to improve sperm quality by reducing oxidative damage. At the same time, there is a growing interest in the literature on the use of testicular sperm in patients with high DNA fragmentation index (DFI). This narrative review aims to evaluate the effectiveness of supplementation of oral antioxidants in infertile men with high DFI compared to testicular sperm retrieval. The current evidence is non-conclusive because of serious risk of bias due to small sample sizes and statistical methods. Further large well-designed randomised placebo-controlled trials are still required to clarify the exact role of these to different therapeutic approaches.


Assuntos
Antioxidantes , Infertilidade Masculina , Humanos , Masculino , Antioxidantes/uso terapêutico , Fragmentação do DNA , Infertilidade Masculina/tratamento farmacológico , Infertilidade Masculina/etiologia , Sêmen , Espermatozoides , Fertilidade
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