RESUMO
OBJECTIVE: To investigate the metabolomic signature of trisomy 21 preimplantation human embryos by a noninvasive approach using mass spectrometry- (MS-) and nuclear magnetic resonance spectroscopy- (NMR-) based metabolic profiling platforms. DESIGN: A total of 171 spent media samples were collected from day 3 embryos and comparatively analyzed by MS analysis (chromosomally normal embryos, n = 15; trisomy 21 embryos, n = 15) and a matched control media group (without embryo, n = 14) and by NMR spectroscopy (normal embryos, n = 39; trisomy 21 embryos, n = 35; monosomy 21 embryos, n = 24) and a matched control media group (without embryo, n = 29). SETTING: IVF clinic/preimplantation genetic diagnosis (PGD) unit facilities. PATIENT(S): One hundred seventy-one spent media samples obtained from human IVF embryos from patients included in our PGD program. INTERVENTION(S): Metabolomic profiling of embryo spent media using liquid chromatography/gas chromatography coupled with MS and NMR. MAIN OUTCOME MEASURE(S): Comparative identification of the metabolites present in the spent media from normal versus trisomy/monosomy 21 day 3 embryos. RESULT(S): Two metabolites, caproate and androsterone sulphate, and two unknown compounds were differentially expressed between normal and trisomy 21 day 3 embryos. Furthermore, the NMR results indicate that there could be a correlation between the differences found between trisomy 21/monosomy 21 and the normal embryos in a spectral region compatible with isoleucine. CONCLUSION(S): This study suggests that the use of differential metabolomic markers found in spent media from preimplantation embryos could be a feasible method for the detection of aneuploidies before ET.
Assuntos
Blastocisto/metabolismo , Síndrome de Down/diagnóstico , Fertilização in vitro/efeitos adversos , Metabolômica , Diagnóstico Pré-Implantação/métodos , Androsterona/análogos & derivados , Androsterona/metabolismo , Aneuploidia , Biomarcadores/metabolismo , Caproatos/metabolismo , Estudos de Casos e Controles , Cromatografia Líquida , Meios de Cultura/metabolismo , Síndrome de Down/genética , Síndrome de Down/metabolismo , Técnicas de Cultura Embrionária , Estudos de Viabilidade , Feminino , Cromatografia Gasosa-Espectrometria de Massas , Humanos , Hibridização in Situ Fluorescente , Espectroscopia de Ressonância Magnética , Masculino , Valor Preditivo dos Testes , Gravidez , Análise para Determinação do SexoRESUMO
A prospective study was performed to assess the impact of sperm DNA fragmentation on the outcome of IVF with own or donated oocytes. The study population included 178 couples (62 cycles of IVF, 116 of intracytoplasmic sperm injection (ICSI)) with own (n=77) and donor (n=101) oocytes. DNA fragmentation was evaluated by TdT (terminal deoxynucleotidyl transferase)-mediated dUDP nick-end labelling assay. Correlation between DNA damage to oocyte fertilization, embryo quality and clinical pregnancy, implantation and miscarriage rates was evaluated. DNA fragmentation was not related to fertilization rates in either IVF (r=0.08) or ICSI (r=-0.04) cycles. DNA fragmentation was similar in patients with <50% embryo utilization rate compared with ≥50%, in cancelled and in embryo transfer cycles and in miscarriages and in successful deliveries. Moreover, DNA fragmentation was similar in pregnant and non-pregnant women as well as in IVF with own or donor oocytes. In the multivariable analysis, the odds ratio of DNA after controlling by age was 1.0. Using a 36% sperm fragmentation threshold, results did not vary. It is concluded that DNA damage was not related to outcomes of IVF or ICSI with own or donor oocytes.
Assuntos
Fragmentação do DNA , Fertilização in vitro , Resultado da Gravidez/genética , Espermatozoides , Feminino , Fertilização , Humanos , Masculino , Análise Multivariada , Doação de Oócitos , Oócitos , Gravidez , Taxa de GravidezRESUMO
Rats were treated with 0, 8, 16 and 24 mg/kg of lead acetate (LA) (i.p.) for 35 days with or without Maca. Maca was co-administrated orally from day 18 to day 35. The lengths of stages of the seminiferous epithelium were assessed by transillumination. Also, sex organ weights, testicular and epididymal sperm count, sperm motility, daily sperm production, sperm transit rate and serum testosterone levels were measured. Lead acetate treatment resulted in a dose-response reduction of lengths of stages VIII and IX-XI, and serum testosterone levels. However, rats treated with 8 and 16 mg/kg but not 24 mg/kg of lead acetate showed a low number of testicular spermatids, low daily sperm production (DSP) and low epididymal sperm count. Administration of Maca to rats treated with lead acetate resulted in higher lengths of stages VIII and IX-XI with respect to lead acetate-treated rats. Moreover, treatment with Maca to lead acetate-treated rats resulted in lengths of stages VIII and IX-XI similar to the control group. Maca administration also reduced the deleterious effect on DSP caused by lead acetate treatment. Maca prevented LA-induced spermatogenic disruption in rats and it may become in a potential treatment of male infertility associated with lead exposure.
