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1.
Int J Mol Sci ; 21(4)2020 Feb 11.
Artigo em Inglês | MEDLINE | ID: mdl-32054116

RESUMO

Excessive levels of reactive nitrogen species (RNS) produce nitrosative stress. Among RNS is peroxynitrite, a highly reactive free radical generated when nitric oxide reacts with superoxide anion. Peroxynitrite effects have been mainly studied in somatic cells, and in spermatozoa the majority of studies are focused in humans. The aim of this study is to investigate the in vitro peroxynitrite effect on boar spermatozoa functions and the molecular mechanisms involved. Spermatozoa were exposed to the donor 3-morpholinosydnonimine (SIN-1) in non-capacitating or capacitating medium, motility was evaluated by CASA, functional parameters by flow cytometry and sperm protein phosphorylation by Western blotting. SIN-1 treatment, that significantly increases peroxynitrite levels in boar spermatozoa, potentiates the capacitating-stimulated phosphorylation of cAMP-dependent protein kinase 1 (PKA) substrates and GSK-3α. SIN-1 induced peroxynitrite does not decrease sperm viability, but significantly reduces sperm motility, progressive motility, velocities and motility coefficients. Concomitantly, peroxynitrite does not affect mitochondrial membrane potential, plasma membrane fluidity, or A23187-induced acrosome reaction. However, peroxynitrite significantly increases sperm lipid peroxidation in both media. In conclusion, peroxynitrite compromises boar sperm motility without affecting mitochondrial activity. Although peroxynitrite potentiates the phosphorylation of pathways leading to sperm motility, it also causes oxidative stress that might explain, at least partially, the motility impairment.

3.
Reprod Biomed Online ; 39(5): 860-867, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31564650

RESUMO

RESEARCH QUESTION: Does ovarian stimulation for oocyte vitrification affect disease-free survival and overall survival rates in women with early breast cancer? DESIGN: This cohort study included 259 patients with early breast cancer; 148 patients underwent ovarian stimulation, whereas 111 patients did not. Patients were treated between January 2008 and December 2016. To calculate the disease-free survival time and overall survival rate, the time of definitive surgery was defined as the starting point. The follow-up was conducted up to 5 years. RESULTS: Exposed and non-exposed groups were comparable in tumour, node and metastases classification, Nottingham grade, hormonal receptor status, tumour molecular phenotype, histology and pathology stage. The exposed group was younger than the non-exposed. Recurrences occurred in 9/148 women (6.1%) in the exposed group and 15/111 women (13.5%) in the non-exposed group, with no significant difference. The mean disease-free survival time was 63.9 months (95% confidence interval [CI]: 61.5-66.4) in the exposed group and 60.6 months (95% CI: 56.9-64.2) in the non-exposed, with no significant difference (log-rank [Mantel-Cox] test). Overall survival rates were comparable; 2/148 (1.4%) and 4/111 (3.6%) patients died, in exposed and non-exposed groups, respectively, during the period analysed. Mean overall survival times were 67.2 months (95% CI: 66.2-68.2) in the exposed group and 65.9 months (95% CI: 64.0-67.9) in the unexposed, with no significant difference (log-rank [Mantel-Cox] test). CONCLUSIONS: This study suggests that ovarian stimulation in patients with early-stage breast cancer is safe in the long term.

4.
J Assist Reprod Genet ; 36(9): 1867-1875, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31392663

RESUMO

PURPOSE: The aim was to evaluate mtDNA content and its dynamics in euploid and aneuploid embryos from cleavage to blastocyst stage following consecutive biopsies. The effect of female age on mtDNA content was evaluated by comparing reproductively younger (≤ 37 years) with older (> 37 years) women. METHODS: A retrospective single-centre descriptive study was performed between August 2016 and January 2017. Forty patients, with 112 embryos, undergoing preimplantation genetic testing for aneuploidies (PGT-A) by next-generation sequencing (NGS) were included. Embryos that reached the blastocyst stage and were not selected for fresh embryo transfer were included following consecutive biopsies of a single blastomere on day 3 and trophectoderm biopsy of day 5 blastocysts. RESULTS: Cleavage-stage mtDNA was significantly lower in fast cleaving embryos (p = 0.016). Based on the concordance between day 3 and day 5 biopsies, a difference was identified in blastocyst mtDNA content between groups (p = 0.019); true euploid blastocysts presented a lower mtDNA content. No association was identified between cleavage-stage mtDNA content and ploidy status (OR 1.008 [0.981-1.036], p = 0.565) nor between blastocyst mtDNA content and ploidy outcome (OR 0.954 [0.898-1.014], p = 0.129). No difference was found when comparing mtDNA content and ploidy outcome between the two reproductive age groups (p = 0.505 (cleavage stage) and p = 0.774 (blastocyst)). CONCLUSION: Mitochondrial DNA content of cleavage-stage embryos and blastocysts is unable to predict ploidy status. Subgroup analysis based on ploidy concordance between day 3 and day 5 revealed a significantly lower mtDNA content for true euploid blastocysts. Reproductive ageing does not affect mtDNA content.

5.
Urology ; 132: 109-116, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31326545

RESUMO

OBJECTIVE: To determine whether a clinically-relevant change in the total motile sperm count (TMSC) over time exists within the subfertile population. METHODS: The first semen analysis of all men presenting to selected infertility centers in 2 countries between 2002 and 2017 were evaluated. Semen analyses were categorized into 3 clinically-relevant groups based on treatment options: TMSC >15 million (M) (Group 1), in which no insemination intervention would be required; TMSC 5-15 M (Group 2), in which intrauterine insemination would be appropriate; and TMSC of <5 M (Group 3), in which in vitro fertilization would be considered. Relationships between male age, TMSC, trend of TMSC, and TMSC group membership by year were assessed. RESULTS: A total of 119,972 first semen analyses were included. The proportion of men with normal TMSC (>15 M) was found to decline approximately 10 percentage points over the past 16 years in the analysis of combined centers (odds ratio 0.967; 95% confidence interval = 0.963-0.971; P = 2.2e-16). A reciprocal increase was distributed between both the moderate (5-15 M) and severe (<5 M) oligozoospermia groups. Additionally, TMSC declined 1.1 percentage points with each year of advancing paternal age. No difference was seen in age at presentation by year. CONCLUSION: The proportion of men with normozoospermia declined and that of men at risk of requiring fertility treatment increased over the study time period. Although several unknown factors may have influenced our data as a result of the retrospective design, a shift in treatment group membership over time may be clinically relevant.


Assuntos
Infertilidade Masculina/fisiopatologia , Contagem de Espermatozoides , Motilidade Espermática , Adulto , Humanos , Infertilidade Masculina/terapia , Masculino , Estudos Retrospectivos , Análise do Sêmen , Fatores de Tempo
6.
PLoS One ; 13(11): e0206098, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30383804

RESUMO

In recent years there is increasing evidence that elevated progesterone levels during ovarian stimulation for IVF / ICSI have a negative impact on the ART-outcome. However, different progesterone assays were used in the previous studies and different assays might produce varying results. This retrospective study evaluated the reproducibility and reliability of different progesterone assays with a special focus on progesterone levels below 1.5 ng/ml, as this range is crucial for early detection of progesterone rise during ovarian stimulation for IVF. A total of 413 blood samples were categorized in different progesterone ranges and whether they were retrieved on the day of final oocyte maturation and the results were compared regarding their reproducibility and reliability. To compare the reproducibility between the different progesterone assays, the Intraclass Correlation Coefficient (ICC) was calculated and interpretation of the ICC results was done according to Cicchetti, ranging from poor to excellent. The correlation of the assays was excellent when all samples were compared including samples retrieved on day of final oocyte maturation, however in the ranges of progesterone levels 1.0 ng/ml to < 1.5 ng/ml, 0.8 ng/ml to < 1.0 ng/ml and < 0.8 ng/ml, the ICC varied between poor and excellent. The assays "gen III" and "Architect" showed an excellent reproducibility of progesterone results throughout all ranges of progesterone levels. This analysis demonstrates, that different progesterone assays have a limited reproducibility and that the results depend on the assay used and the range of progesterone level. This fact leads to two important conclusions. Firstly the limited reproducibility might lead to substantially different treatment decisions in ovarian stimulation treatment for IVF and secondly critical interpretation of thresholds, provided by meta-analysis, is crucial despite the risk that the so far gained clinical experience might become irrelevant and has to be adjusted to the results, obtained by each assay.


Assuntos
Oócitos/metabolismo , Oogênese/genética , Indução da Ovulação , Progesterona/isolamento & purificação , Adulto , Feminino , Fertilização In Vitro , Humanos , Oócitos/crescimento & desenvolvimento , Gravidez , Taxa de Gravidez , Progesterona/metabolismo , Injeções de Esperma Intracitoplásmicas
7.
Reprod Biomed Online ; 37(4): 415-424, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30396454

RESUMO

RESEARCH QUESTION: Can gonadotrophin releasing hormone (GnRH) antagonist be used in egg donation recipients with ovulatory cycles for the purpose of achieving synchronization between the donor´s and recipient´s cycle? DESIGN: Prospective randomized controlled trial to compare 7-day dosage of GnRH antagonist for endometrial priming in an oocyte donation programme with a single dose of long-acting GnRH agonist. A total of 563 women were randomized in a private single centre, and 473 women underwent embryo transfer. Ongoing pregnancy rate was the primary end point. Analysis was adjusted for embryonic stage at the time of embryo transfer; data collected included days on the waiting list; number of fresh-vitrified oocytes collected; and oocyte donor´s age at the time of retrieval. RESULTS: No statistically significant differences were found between groups in per intention-to-treat analysis: adjusted OR 1.42 (CI 0.97 to 2.09); per treatment received: adjusted OR 1.43 (CI 0.97 to 2.09); per embryo transfer: adjusted OR for ongoing pregnancy rate 1.47 (CI 1.01 to 2.13), P = 0.047. CONCLUSIONS: For women with ovulatory cycles undergoing oocyte donation, the outcomes are similar between GnRH antagonist and down-regulated hormone replacement protocols.


Assuntos
Transferência Embrionária/métodos , Endométrio/efeitos dos fármacos , Hormônio Liberador de Gonadotropina/antagonistas & inibidores , Doação de Oócitos , Adulto , Desenvolvimento Embrionário , Endométrio/fisiologia , Feminino , Fertilização In Vitro , Humanos , Indução da Ovulação/métodos , Gravidez , Taxa de Gravidez
8.
J Assist Reprod Genet ; 35(11): 1973-1985, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30105539

RESUMO

Although assisted reproduction techniques involve the use of semen samples, there is little scientific methodology applied when selecting sperm. To select the most appropriate spermatozoa, first we need to define the optimal molecular characteristics. Sperm lipids may contribute to sperm function, thus our aim was to compare the lipidic profiles of sperm samples used in intracytoplasmic sperm injection cycles that ultimately led to a pregnancy with those that did not.Spermatozoa from infertile patients after intracytoplasmic sperm injection (group non-pregnant, n = 16; vs. group pregnant, n = 22) were analyzed for lipid composition using ultra-high performance liquid chromatography coupled to mass spectrometry, by means two platforms for measuring fatty acyls, bile-acids, lysoglycerophospholipids, glycerolipids, cholesteryl-esters, sphingolipids, and glycerophospholipids. Lipid levels were compared using a univariate test and multivariate analyses after logarithmic transformation.We detected 151 different lipids in the sperm samples, 10 of which were significantly increased in sperm samples from the NP group, ranging from 1.10- to 1.30-fold change. These were primarily ceramides, sphingomyelins and three glycerophospholipids, a lysophosphatidylcholine, and two plasmalogen species. Additionally, 2-Monoacylglycerophosphocholine were also found in higher levels in non-pregnant group.Our results describe the composition of sperm lipids linked to optimal sperm function, opening new possibilities for the development of male fertility diagnostic tools and culture media formulations to improve sperm quality and enhance reproductive results. Given that lipids compose the majority of the sperm plasma membrane, this information is also useful in designing new sperm selection tools that will allow for the selection of the best spermatozoa.


Assuntos
Biomarcadores/sangue , Infertilidade Masculina/sangue , Lipídeos/sangue , Técnicas de Reprodução Assistida , Injeções de Esperma Intracitoplásmicas/métodos , Espermatozoides/metabolismo , Estudos de Casos e Controles , Feminino , Humanos , Infertilidade Masculina/diagnóstico , Infertilidade Masculina/terapia , Masculino , Indução da Ovulação , Gravidez , Resultado da Gravidez , Estudos Prospectivos
9.
Asian J Androl ; 20(5): 498-504, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29873314

RESUMO

Angiotensin-converting enzyme functions in the male reproductive system, but the extent of its function in reproduction is not fully understood. The primary objective of this work was to investigate the relationship between the testicular isoform of angiotensin-converting enzyme present in human spermatozoa and semen parameters, human embryo quality, and assisted reproduction success. A total of 81 semen samples and 635 embryos from couples undergoing oocyte donation cycles at the IVI Bilbao Clinic were analyzed. Semen parameters, embryos quality, and blastocyst development were examined according to the World Health Organization standards and the Spanish Association of Reproduction Biology Studies criteria. The percentage of testicular angiotensin-converting enzyme-positive spermatozoa and the number of molecules per spermatozoon were analyzed by flow cytometry. Both parameters were inversely correlated with human sperm motility. Higher percentages of testicular angiotensin-converting enzyme-positive spermatozoa together with fewer enzyme molecules per spermatozoon were positively correlated with better embryo quality and development. Our results suggest that embryos with a higher implantation potential come from semen samples with higher percentages of testicular angiotensin-converting enzyme-positive cells and fewer enzyme molecules per spermatozoon. Based on these findings, we propose that testicular angiotensin-converting enzyme could be used to aid embryologists in selecting better semen samples for obtaining high-quality blastocysts during in vitro fertilization procedures.


Assuntos
Desenvolvimento Embrionário/fisiologia , Peptidil Dipeptidase A/metabolismo , Motilidade Espermática/fisiologia , Espermatozoides/enzimologia , Testículo/enzimologia , Adulto , Implantação do Embrião/fisiologia , Transferência Embrionária , Fertilidade/fisiologia , Fertilização In Vitro , Humanos , Masculino , Pessoa de Meia-Idade
10.
J Turk Ger Gynecol Assoc ; 19(1): 48-52, 2018 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-29503262

RESUMO

Human papilloma virus (HPV) is one of the most common viral sexually-transmitted diseases worldwide. The prevalence of HPV is higher in infertile males when compared with fertile men and ranges between 10 and 35.7% in men affected by unexplained infertility. HPV can bind to spermatozoa and can potentially be transferred to fertilized oocytes. Viral detection in blastocysts and trophoblastic cells is associated with impaired embryo development and poor pregnancy outcomes. Nevertheless, attempts to eliminate HPV-DNA from sperm samples through routine washing techniques have failed. In assisted reproduction technologies (ART), intracytoplasmic sperm injection involves no natural selection of the sperm cell, which means that these procedures have a plausible risk of injecting sperm containing HPV. The possible detrimental effects of HPV on ART in couples with infected male partners are summarized in this review.

11.
Artigo em Inglês | MEDLINE | ID: mdl-29497400

RESUMO

Ovarian stimulation in a gonadotropin-releasing hormone (GnRH) antagonist protocol with the use of GnRH agonist for final oocyte maturation is the state-of-the-art treatment in patients with an expected or known high response to avoid or at least reduce significantly the risk for development of ovarian hyperstimulation syndrome (OHSS). Due to a shortened LH surge after administration of GnRH agonist in most patients, the luteal phase will be characterized by luteolysis and luteal phase insufficiency. Maintaining a sufficient luteal phase is crucial for achievement of a pregnancy; however, the optimal approach is still under debate. Administration of human chorionic gonadotropin (hCG) within 72 h rescues the corpora lutea function; however, the so far often used 1,500 IU still bear the risk for development of OHSS. The recently introduced concept of "luteal coasting" individualizes the luteal phase support by monitoring the progesterone concentrations and administering a rescue dosage of hCG when progesterone concentrations drop significantly. This retrospective proof-of-concept study explored the correlation between hCG dosages ranging from 375 up to 1,500 IU and the progesterone levels in the early and mid-luteal phases as well as the likelihood of pregnancy, both early and ongoing. The chance of pregnancy is highest with progesterone level ≥13 ng/ml at 48 h postoocyte retrieval. Among the small sample size of 52 women studied, it appears that appropriate progesterone levels can be achieved with hCG dosages as low as 375 IU. This may well optimize the chance of pregnancy while reducing the risk of OHSS associated with higher doses of hCG supplementation in the luteal phase.

12.
Asian J Androl ; 20(1): 1-8, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-28440264

RESUMO

Spermatozoa retrieved from the testis of men with high levels of sperm DNA fragmentation (SDF) in the neat semen tend to have better DNA quality. Given the negative impact of SDF on the outcomes of Assisted Reproductive Technology (ART), an increased interest has emerged about the use of testicular sperm for intracytoplasmic sperm injection (Testi-ICSI). In this article, we used a SWOT (strengths, weaknesses, opportunities, and threats) analysis to summarize the advantages and drawbacks of this intervention. The rationale of Testi-ICSI is bypass posttesticular DNA fragmentation caused by oxidative stress during sperm transit through the epididymis. Hence, oocyte fertilization by genomically intact testicular spermatozoa may be optimized, thus increasing the chances of creating a normal embryonic genome and the likelihood of achieving a live birth, as recently demonstrated in men with high SDF. However, there is still limited evidence as regards the clinical efficacy of Testi-ICSI, thus creating opportunities for further confirmatory clinical research as well as investigation of Testi-ICSI in clinical scenarios other than high SDF. Furthermore, Testi-ICSI can be compared to other laboratory preparation methods for deselecting sperm with damaged DNA. At present, the available literature supports the use of testicular sperm when performing ICSI in infertile couples whose male partners have posttesticular SDF. Due to inherent risks of sperm retrieval, Testi-ICSI should be offered when less invasive treatments for alleviating DNA damage have failed. A call for continuous monitoring is nonetheless required concerning the health of generated offspring and the potential complications of sperm retrieval.


Assuntos
Fragmentação do DNA , Injeções de Esperma Intracitoplásmicas/métodos , Espermatozoides , Testículo/citologia , Adulto , Feminino , Humanos , Infertilidade Masculina , Masculino , Gravidez , Resultado da Gravidez , Recuperação Espermática
14.
Epigenomics ; 9(10): 1299-1315, 2017 10.
Artigo em Inglês | MEDLINE | ID: mdl-28877596

RESUMO

AIM: To characterize the sperm methylome in semen samples from 19 donors with proven fertility. MATERIALS & METHODS: Bisulfite-converted sperm DNA was hybridized on the HumanMethylation450 Infinium BeadChip platform. CpG fluorescence intensities were extracted and converted to ß-values. RESULTS: The sperm methylome is highly homogeneous and hypomethylated. Genes with hypomethylated promoters are ontologically associated to biological functions related to spermatogenesis and embryogenesis. Sex chromosomes are the most hypomethylated chromosomes, supporting data that indicated their essential role in spermatogenesis. A total of 94 genes are resistant to demethylation, being strong candidates for transgenerational inheritance. CONCLUSION: Spermatozoa carry a homogeneous methylation profile that is a footprint of past events (spermatogenesis), is designed to facilitate future events (embryogenesis) and has a possible influence in the adult life (transgenerational effects).


Assuntos
Metilação de DNA , Epigênese Genética , Espermatozoides/metabolismo , Adulto , Cromossomos Humanos X/genética , Cromossomos Humanos Y/genética , Ilhas de CpG , Desenvolvimento Embrionário/genética , Humanos , Masculino , Regiões Promotoras Genéticas , Espermatogênese/genética , Espermatozoides/citologia
15.
Fertil Steril ; 108(3): 456-467.e1, 2017 09.
Artigo em Inglês | MEDLINE | ID: mdl-28865546

RESUMO

OBJECTIVE: To compare sperm DNA fragmentation (SDF) levels between testicular and ejaculated sperm and to evaluate outcomes of intracytoplasmic sperm injection (ICSI) with the use of testicular (Testi-ICSI) versus ejaculated (Ejac-ICSI) sperm in nonazoospermic men with high SDF. DESIGN: Systematic review and meta-analysis. SETTING: Not applicable. PATIENT(S): Normo- and oligozoospermic men with high levels of SDF in semen subjected to Testi-ICSI or Ejac-ICSI. INTERVENTION(S): Summary mean difference (MD) and odds ratio (OR) were calculated with the use of an inverse variance model and fixed- or random-effects models, respectively. MAIN OUTCOME MEASURE(S): Primary outcomes were SDF levels, clinical pregnancy rates (CPRs), and live birth rates (LBRs). Secondary outcomes were fertilization and miscarriage rates. RESULT(S): Five studies involving 143 patients provided paired SDF rates for testicular and ejaculated sperm, revealing lower SDF in testicular sperm (MD -24.58%). Four studies involving 507 cycles and 3,840 oocytes reported clinical outcomes of Testi-ICSI and Ejac-ICSI. Fertilization rates were not different between sperm sources, but a trend to lower rates was observed with Testi-ICSI. CPRs were higher for Testi-ICSI than for Ejac-ICSI, as were LBRs, whereas miscarriage rates were reduced with Testi-ICSI. CONCLUSION(S): Testicular sperm have lower levels of SDF than ejaculated sperm, with Testi-ICSI for high post-testicular SDF men improving reproductive outcomes compared with Ejac-ICSI. Infertile couples may benefit from Testi-ICSI if male partners have confirmed high SDF in the ejaculate.


Assuntos
Ejaculação , Infertilidade/epidemiologia , Infertilidade/terapia , Resultado da Gravidez/epidemiologia , Injeções de Esperma Intracitoplásmicas/estatística & dados numéricos , Recuperação Espermática/estatística & dados numéricos , Testículo/metabolismo , Adulto , Fragmentação do DNA , Feminino , Humanos , Masculino , Gravidez , Injeções de Esperma Intracitoplásmicas/métodos , Resultado do Tratamento , Adulto Jovem
16.
PLoS One ; 12(5): e0176600, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28459828

RESUMO

Final oocyte maturation using GnRH-agonist trigger in a GnRH-antagonist protocol is increasingly common, as ovarian hyperstimulation syndrome is almost completely avoided. However, this approach might lead to reduced pregnancy rates due to severe luteolysis. This proof of concept study evaluated the extend of luteolysis by measuring progesterone levels 48 hours after oocyte retrieval in 51 patients, who received GnRH-agonist trigger for final oocyte maturation in a GnRH-antagonist protocol due to the risk of ovarian hyperstimulation syndrome. It was shown, that luteolysis after GnRHa-trigger differs greatly among patients, with progesterone levels ranging from 13.0 ng/ml to ≥ 60.0 ng/ml, 48 hours after oocyte retrieval. Significant positive correlations could be demonstrated between progesterone levels and the number of ovarian stimulation and suppression days (p = 0.006 and p = 0.002 respectively), the total amount of medication used for ovarian suppression (p = 0.015), the level of progesterone on the day of final oocyte maturation (p = 0.008) and the number of retrieved oocytes (p = 0.019). Therefore it was concluded, that luteolysis after GnRH-agonist trigger is patient-specific and also luteal phase support requires individualization. Longer stimulation duration as well as a higher level of progesterone on the day of final oocyte maturation and more retrieved oocytes will result in higher levels of progesterone 48 hours after oocyte retrieval.


Assuntos
Fármacos para a Fertilidade Feminina/farmacologia , Hormônio Liberador de Gonadotropina/agonistas , Luteólise/efeitos dos fármacos , Oócitos/efeitos dos fármacos , Progesterona/metabolismo , Adulto , Feminino , Fármacos para a Fertilidade Feminina/uso terapêutico , Antagonistas de Hormônios/farmacologia , Antagonistas de Hormônios/uso terapêutico , Humanos , Infertilidade Feminina/metabolismo , Infertilidade Feminina/terapia , Fase Luteal/efeitos dos fármacos , Fase Luteal/metabolismo , Luteólise/fisiologia , Recuperação de Oócitos , Oócitos/fisiologia , Oogênese/efeitos dos fármacos , Oogênese/fisiologia , Indução da Ovulação , Adulto Jovem
17.
J Assist Reprod Genet ; 34(2): 201-207, 2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-27882439

RESUMO

PURPOSE: The purpose of this study is to assess outcomes after magnetic-activated cell sorting (MACS) technology on obstetric and perinatal outcomes compared with those achieved after swim up from randomized controlled trial. METHODS: This is a two-arm, unicentric, prospective, randomized, and triple-blinded trial and has a total of 237 infertile couples, between October 2010 and January 2013. A total of 65 and 66 newborns from MACS and control group, respectively, were described. RESULTS: MACS had no clinically relevant adverse effects on obstetric and perinatal outcomes. No differences were found for obstetric problems including premature rupture of membranes 6.1% (CI95% 0-12.8) vs. 5.9% (CI95% 0-12.4), 1st trimester bleeding 28.6% (CI95% 15.9-41.2) vs. 23.5% (CI95% 11.9-35.1), invasive procedures as amniocentesis 2.0% (CI95% 0-5.9) vs. 3.9% (CI95% 0-9.2), diabetes 14.3% (CI95% 4.5-24.1) vs. 9.8% (CI95% 1.6-17.9), anemia 6.1% (CI95% 0-12.8) vs. 5.9%(CI95% 0-12.4), 2nd and 3rd trimesters 10.2% (CI95% 1.7-18.7) vs. 5.9% (CI95% 0-12.4), urinary tract infection 8.2% (CI95% 0.5-15.9) vs. 3.9% (CI95% 0-9.2), pregnancy-induced hypertension 6.1% (CI95% 0-12.8) vs. 15.7% (CI95% 5.7-25.7), birth weight (g) 2684.10 (CI95% 2499.48-2868.72) vs. 2676.12 (CI95% 2499.02-2852.21), neonatal height (cm) 48.3 (CI95% 47.1-49.4) vs. 46.5 (CI95% 44.6-48.4), and gestational cholestasis 0%(CI95% 0-0) vs. 3.9% (CI95% 0-9.2), respectively, in MACS group compared with control group. CONCLUSIONS: Our data suggest that MACS technology does not increase or decrease Powered by Editorial Manager® and ProduXion Manager® from Aries Systems Corporation adverse obstetric and perinatal outcomes in children conceived when this technology was performed, being the largest randomized control trial with live birth reported results with MACS.


Assuntos
Fertilização In Vitro , Infertilidade/patologia , Complicações na Gravidez/patologia , Espermatozoides/crescimento & desenvolvimento , Adulto , Peso ao Nascer , Separação Celular/métodos , Colestase Intra-Hepática/patologia , Feminino , Citometria de Fluxo/métodos , Humanos , Hipertensão Induzida pela Gravidez/patologia , Recém-Nascido , Masculino , Gravidez , Resultado da Gravidez , Nascimento Prematuro
18.
Reprod Biomed Online ; 33(6): 709-719, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-27692602

RESUMO

The influence of aberrant sperm DNA methylation on the reproductive capacity of couples has been postulated as a cause of infertility. This study compared the DNA methylation of spermatozoa of 19 fertile donors and 42 infertile patients using the Illumina 450K array. Clustering analysis of methylation data arranged fertile and infertile patients into two groups. Bivariate clustering analysis identified a differential distribution of samples according to the characteristics of seminogram and age, suggesting a possible link between these parameters and specific methylation profiles. The study identified 696 differentially methylated cytosine-guanine dinucleotides (CpG) associated with 501 genes between fertile donors and infertile patients. Ontological enrichment analysis revealed 13 processes related to spermatogenesis. Data filtering identified a set of 17 differentially methylated genes, some of which had functions relating to spermatogenesis. A significant association was identified between RPS6KA2 hypermethylation and advanced age (P = 0.016); APCS hypermethylation and oligozoospermia (P = 0.041); JAM3/NCAPD3 hypermethylation and numerical chromosome sperm anomalies (P = 0.048); and ANK2 hypermethylation and lower pregnancy rate (P = 0.040). This description of a set of differentially methylated genes provides a framework for further investigation into the influence of such variation in male fertility in larger patient cohorts.


Assuntos
Metilação de DNA , Infertilidade Masculina/genética , Espermatogênese/genética , Espermatozoides/metabolismo , Adulto , Análise por Conglomerados , Ilhas de CpG , Feminino , Fertilidade/genética , Humanos , Masculino , Oligospermia/genética , Gravidez , Taxa de Gravidez , Regiões Promotoras Genéticas , Reprodução , Adulto Jovem
19.
J Proteome Res ; 15(11): 4101-4115, 2016 11 04.
Artigo em Inglês | MEDLINE | ID: mdl-27581094

RESUMO

The current catalogue of the human proteome is not yet complete, as experimental proteomics evidence is still elusive for a group of proteins known as the missing proteins. The Human Proteome Project (HPP) has been successfully using technology and bioinformatic resources to improve the characterization of such challenging proteins. In this manuscript, we propose a pipeline starting with the mining of the PRIDE database to select a group of data sets potentially enriched in missing proteins that are subsequently analyzed for protein identification with a method based on the statistical analysis of proteotypic peptides. Spermatozoa and the HEK293 cell line were found to be a promising source of missing proteins and clearly merit further attention in future studies. After the analysis of the selected samples, we found 342 PSMs, suggesting the presence of 97 missing proteins in human spermatozoa or the HEK293 cell line, while only 36 missing proteins were potentially detected in the retina, frontal cortex, aorta thoracica, or placenta. The functional analysis of the missing proteins detected confirmed their tissue specificity, and the validation of a selected set of peptides using targeted proteomics (SRM/MRM assays) further supports the utility of the proposed pipeline. As illustrative examples, DNAH3 and TEPP in spermatozoa, and UNCX and ATAD3C in HEK293 cells were some of the more robust and remarkable identifications in this study. We provide evidence indicating the relevance to carefully analyze the ever-increasing MS/MS data available from PRIDE and other repositories as sources for missing proteins detection in specific biological matrices as revealed for HEK293 cells.


Assuntos
Biologia Computacional/métodos , Bases de Dados de Proteínas , Proteoma/análise , Aorta/química , Feminino , Lobo Frontal/química , Células HEK293 , Humanos , Masculino , Placenta/química , Gravidez , Proteômica/métodos , Retina/química , Espermatozoides/química , Espectrometria de Massas em Tandem
20.
Fertil Steril ; 105(3): 608-616, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26616438

RESUMO

OBJECTIVE: To investigate the presence of angiotensin II type 2 receptor (AT2R) in human spermatozoa and its implication in sperm fertility status. DESIGN: We carried out expression assays for AT2R by reverse transcription-polymerase chain reaction, Western blot, immunofluorescence, and flow cytometry techniques in human sperm cells. Percentage of AT2R-positive sperm cells was analyzed by flow cytometry. SETTING: Assisted reproduction unit and academic research laboratory. PATIENT(S): Ninety-seven human semen samples from the Clínica IVI Bilbao. INTERVENTION(S): All samples were examined and classified according to World Health Organization guidelines. Spermatozoa were isolated from semen on discontinuous colloidal silica gradient (45%-90%) and swim-up techniques. MAIN OUTCOME MEASURE(S): Presence and location of the AT2R in spermatozoa and percentage of AT2R-positive sperm cells measured by flow cytometry. RESULT(S): We demonstrated the existence of AT2R and its transcript in human sperm by Western blot and reverse transcription-polymerase chain reaction. Immunofluorescence studies showed that AT2R is mainly located at the equatorial segment of the sperm head. The AT2R levels were associated with sperm motility parameters. Particularly, we found a significant positive correlation between AT2R and spermatozoa with progressive motility grade and a significant negative correlation with immotile spermatozoa, both in fresh semen samples and in prepared sperm cells. Regarding pathologic studies, the levels of AT2R measured by flow cytometry were lower in spermatozoa of asthenozoospermic men than in normozoospermic controls. CONCLUSION(S): Angiotensin II type 2 receptor is present in human semen and may be involved in the control of sperm motility. In-depth understanding of the proteins involved in sperm motility can help to elucidate the role of these proteins in male infertility as well as to establish new biomarkers for male infertility.


Assuntos
Astenozoospermia/metabolismo , Receptor Tipo 2 de Angiotensina/metabolismo , Motilidade Espermática , Espermatozoides/metabolismo , Astenozoospermia/genética , Astenozoospermia/patologia , Astenozoospermia/fisiopatologia , Biomarcadores/metabolismo , Western Blotting , Estudos de Casos e Controles , Fertilidade , Citometria de Fluxo , Imunofluorescência , Humanos , Masculino , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Receptor Tipo 2 de Angiotensina/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais , Espermatozoides/patologia
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