RESUMEN
BACKGROUND: There is an association between aging ana an increased number of sperms with alterations in nuclear DNA. AIM: To study the association between age and fragmentation of sperm DNA. MATERIAL AND METHODS: Sixty two volunteers provided semen for analysis. These were separated in a group aged less than forty years and a second group aged more than forty years. Sperm DNA fragmentation was studied by TUNEL (terminal deoxynucleotidyl transferase-mediated 2'-deoxyuridine 5'-triphosphate nick end-labeling) and SCD (sperm chromatin dispersion test) assays. RESULTS: Compared with their younger counterparts, patients aged more than 40 years had a higher proportion of sperms with DNA fragmentation by TUNEL (20 ± 1.3 and 24 ± 1.9% respectively, p < 0.05) and SCD (22 ± 1.4 and 26 ± 1.6% respectively, p < 0.05). The results of both assays had a correlation coefficient of O.8. No differences between groups were observed for other seminal parameters. CONCLUSIONS: Sperm DNA fragmentation increases with age in males.
Asunto(s)
Cromatina/química , Fragmentación del ADN , Etiquetado Corte-Fin in Situ/métodos , Análisis de Semen/métodos , Espermatozoides/ultraestructura , Adolescente , Adulto , Factores de Edad , Humanos , Masculino , Persona de Mediana Edad , Recuento de Espermatozoides , Motilidad Espermática , Espermatozoides/fisiología , Adulto JovenRESUMEN
Background: There is an association between aging ana an increased number ofsperms with alterations in nuclear DNA. Aim: To study the association between age and fragmentation of sperm DNA. Material andMethods: Sixty two volunteers provided semen for analysis. These were separated in a group aged less than forty years and a second group aged more than forty years. Sperm DNA fragmentation was studied by TÚNEL (terminal deoxynucleotidyl transferase-mediated2'-deoxyuridine 5'-triphosphate nick end-labeling) and SCD (sperm chromatin dispersión test) assays. Results: Compared with theiryounger counterparts, patients aged more than 40years had a higher proportion ofsperms with DNA fragmentation by TÚNEL (20 ±1.3 and24 ± 1.9 percent respectively, p < 0.05) and SCD (22 ± 1.4 and26 ± 1.6 percent respectively, p < 0.05). The results ofboth assays had a correlation coefficientofO.8. No differences between groups were observed for other seminal parameters. Conclusions: Sperm DNA fragmentation increases with age in males.
Asunto(s)
Adolescente , Adulto , Humanos , Masculino , Persona de Mediana Edad , Adulto Joven , Cromatina/química , Fragmentación del ADN , Etiquetado Corte-Fin in Situ/métodos , Análisis de Semen/métodos , Espermatozoides/ultraestructura , Factores de Edad , Recuento de Espermatozoides , Motilidad Espermática , Espermatozoides/fisiologíaRESUMEN
OBJECTIVE: To determine whether the incidence of apoptosis in mature oocyte cumulus cells changes after insemination related to infertility. DESIGN: Prospective study. SETTING: Public hospital and university. PATIENT(S): One hundred women undergoing in vitro fertilization and embryo transfer (IVF-ET). INTERVENTION(S): Collection of cumulus cells from IVF-ET cycles with different infertility etiologies. MAIN OUTCOME MEASURE(S): Detection of apoptosis in cumulus cells; fertilization, embryo quality, and pregnancy rate. RESULT(S): The incubation of cumulus-oocyte-complexes with spermatozoa led to an increase in cumulus cell apoptosis from 34.2 +/- 3.7 to 44.5 +/- 6.3%. After insemination, cumulus cells of poor quality embryos showed a statistically higher apoptotic rate versus cumulus cells of good quality embryos (61.5 +/- 6.4 vs. 40.6 +/- 3.9%). Cumulus cells arising from oocytes with >or=50% fertilization rates after insemination showed higher apoptosis rates did cumulus cells from oocytes with <50% fertilization rates (46.0 +/- 3.7 vs. 33.8 +/- 4.0%). Patients with endometriosis presented higher apoptotic rates before insemination (77.6 +/- 9.06%). Cumulus cells obtained after aspiration showed no differences in their apoptosis rates for the following factors: age of women, aspirated oocytes, estradiol level, fertilization rate, and embryo quality or pregnancy. The apoptotic profile from pregnant women was less than (but not statistically significantly different from) profiles from nonpregnant women. CONCLUSION(S): These results suggest that the incidence of apoptosis in cumulus cells is associated with exposure to spermatozoa and the cause of infertility.
Asunto(s)
Apoptosis/fisiología , Células del Cúmulo/citología , Oocitos/fisiología , Espermatozoides/fisiología , Adulto , Células del Cúmulo/fisiología , Transferencia de Embrión/métodos , Endometriosis/patología , Femenino , Fertilización , Humanos , Infertilidad Femenina/patología , Infertilidad Femenina/fisiopatología , Masculino , Embarazo , Inyecciones de Esperma Intracitoplasmáticas/métodos , Motilidad EspermáticaRESUMEN
The reproductive system is extremely susceptible to insults from exposure to exogenous steroids during development. Excess prenatal testosterone exposure programs neuroendocrine, ovarian, and metabolic deficits in the female, features seen in women with polycystic ovary disease. The objective of this study was to determine whether prenatal testosterone excess also disrupts the male reproductive system, using sheep as a model system. The extent of reproductive disruption was tested by assessing sperm quantity and quality as well as Leydig cell responsiveness to human chorionic gonadotropin. Males born to mothers treated with 30 mg testosterone propionate twice weekly from d 30 to 90 and with 40 mg testosterone propionate from d 90 to 120 of pregnancy (T-males) showed a significant reduction (P < 0.05) in body weight, scrotal circumference, and sperm count compared with control males. Mean straight line velocity of sperms was also lower in T-males (P < 0.05). Circulating testosterone levels in response to the human chorionic gonadotropin did not differ between groups. These findings demonstrate that exposure to excess testosterone during fetal development has a negative impact on reproductive health of the male offspring, raising concerns relative to unintended human exposure to steroidal mimics in the environment.
Asunto(s)
Efectos Tardíos de la Exposición Prenatal/fisiopatología , Recuento de Espermatozoides , Motilidad Espermática/efectos de los fármacos , Testosterona/toxicidad , Animales , Peso Corporal/efectos de los fármacos , Gonadotropina Coriónica/farmacología , Femenino , Humanos , Células Intersticiales del Testículo/citología , Células Intersticiales del Testículo/efectos de los fármacos , Masculino , Embarazo , Efectos Tardíos de la Exposición Prenatal/inducido químicamente , Maduración Sexual/efectos de los fármacos , Maduración Sexual/fisiología , Ovinos , Propionato de Testosterona/toxicidadRESUMEN
CONTEXT: An important proportion of male members of polycystic ovary syndrome (PCOS) families exhibit insulin resistance and related metabolic defects. However, the reproductive phenotypes in first-degree male relatives of PCOS women have been described less often. OBJECTIVE: The objective of the study was to evaluate the pituitary-testicular function in sons of women with PCOS during different stages of life: early infancy, childhood, and adulthood. DESIGN: Eighty sons of women with PCOS (PCOS(S)) and 56 sons of control women without hyperandrogenism (C(S)), matched for age, were studied. In all subjects, the pituitary-gonadal axis was evaluated by a GnRH agonist test (leuprolide acetate, 10 microg/kg sc). Serum anti-Müllerian hormone (AMH) and inhibin B were used as Sertoli cell markers. Serum concentrations of gonadotropins, steroid hormones, and SHBG were also determined. A semen analysis was performed. RESULTS: Basal concentrations of gonadotropins, sex steroids, and inhibin B were comparable between PCOS(s) and C(S) during early infancy, childhood, and adulthood. Similar results in stimulated gonadotropin and sex steroid concentrations were observed. However, AMH serum concentrations were higher in PCOS(s) compared with C(S) during early infancy [925.0 (457.3-1401.7) vs. 685.6 (417.9-1313.2) pmol/liter, P = 0.039] and childhood [616.3 (304.6-1136.9) vs. 416.5 (206.7-801.2) pmol/liter, P = 0.007). Sperm-count analysis was similar between both groups. CONCLUSIONS: AMH concentrations are increased in prepubertal sons of women with PCOS, suggesting that these boys may show an increased Sertoli cell number or function during infancy and childhood. However, this does not seem to have a major deleterious effect on sperm production.
Asunto(s)
Hijos Adultos , Desarrollo Infantil/fisiología , Hijo de Padres Discapacitados , Hipófisis/fisiología , Síndrome del Ovario Poliquístico , Testículo/fisiología , Adolescente , Adulto , Niño , Preescolar , Femenino , Hormonas Esteroides Gonadales/sangre , Humanos , Lactante , Masculino , Motilidad Espermática/fisiología , Espermatogénesis/fisiologíaRESUMEN
CONTEXT: Polycystic ovary syndrome (PCOS) is a common endocrine-metabolic disorder with strong familial aggregation. It has been demonstrated that parents and brothers of PCOS women exhibit insulin resistance and related metabolic defects. However, metabolic phenotypes in sons of PCOS women have not been described. OBJECTIVE: Our objective was to assess the metabolic profiles in sons of women with PCOS during different stages of life: early infancy, childhood, and adulthood. DESIGN: Eighty sons of women with PCOS (PCOS(S)) and 56 sons of control women without hyperandrogenism (C(S)), matched for age, were studied. In early infancy, glucose and insulin were determined in the basal sample. In children and adults, a 2-h oral glucose tolerance test was performed with measurements of glucose and insulin. Adiponectin, leptin, C-reactive protein, SHBG, and serum lipids were determined in the basal sample during the three periods. RESULTS: During early infancy, PCOS(S) showed higher weight (P = 0.038) and weight sd score (P = 0.031) than C(S). During childhood, weight (P = 0.003), body mass index (BMI) (P < 0.001), BMI sd score (P < 0.001), waist circumference (P = 0.001), total cholesterol (P = 0.007), and low-density lipoprotein cholesterol (P = 0.022) were higher in PCOS(S) compared with C(S), but after adjusting for BMI, these differences were nonsignificant. During adulthood, PCOS(S) exhibited higher weight (P = 0.022), BMI (P = 0.046), and waist circumference (P = 0.028) than C(S). Fasting insulin (P = 0.030), homeostasis model assessment for insulin resistance (P = 0.034), total cholesterol (P = 0.043), low-density lipoprotein cholesterol (P = 0.034), and 2-h insulin (P = 0.006) were also significantly higher and insulin sensitivity index composite significantly lower in PCOS(S) than in C(S) (P = 0.003). After adjusting for BMI, only 2-h insulin and insulin sensitivity index composite remained significantly different. CONCLUSIONS: This study indicates that sons of PCOS women exhibit higher body weight from early infancy. In addition, insulin resistance became evident as the subjects got older, which may place them at risk for the development of type 2 diabetes and cardiovascular disease.
Asunto(s)
Resistencia a la Insulina , Síndrome del Ovario Poliquístico/genética , Adolescente , Adulto , Índice de Masa Corporal , Proteína C-Reactiva/análisis , Niño , Preescolar , LDL-Colesterol/sangre , Familia , Femenino , Humanos , Lactante , Modelos Lineales , Masculino , Obesidad/etiologíaRESUMEN
Exposure of either gametes or embryos to conditions and/or factors that generate oxidative stress has been associated with impaired early embryogenesis. The effects of reactive oxygen species (ROS) on mouse preimplantation development, depending of the ROS-concentration and time of exposition, were studied. Two-cell embryos were incubated with 5, 10, 25 and 50 microM of hydrogen peroxide (H2O2) for 30 and 60 minutes of exposition and allowed to develop for 72 h to study the quality of development. The incubation with 50 microM H2O2 for 30 or 60 minutes, strongly inhibited the 2-cell embryo development as compared to the control (p < 0.001). Twenty-five microM H2O2 produced inhibition of blastocyst formation (p < 0.001) and 10 microM H2O2 significantly decreased the percentages of expanded and hatched blastocysts, which resulted morphologically altered (p < 0.05 and p < 0.01, respectively). The higher H2O2 concentrations were able to elicit necrotic morphology in the 2-cell arrested embryos, while 10 microM H2O2 induced moderate damage with the arrested embryos partially fragmented. In conclusion, important causes for defective preimplantation development and for early embryo losses may be due to oxidative stress because early mouse embryos exposed to ROS for short times arrested at the first cellular cycle (2-cell) and/or impaired embryo differentiation and morphogenesis, being these effects ROS-concentration-dependent.
Asunto(s)
Blastocisto/citología , Blastocisto/efectos de los fármacos , Desarrollo Embrionario/efectos de los fármacos , Peróxido de Hidrógeno/toxicidad , Especies Reactivas de Oxígeno/toxicidad , Animales , Blastocisto/metabolismo , Fase de Segmentación del Huevo/efectos de los fármacos , Transferencia de Embrión , Femenino , Peróxido de Hidrógeno/administración & dosificación , Masculino , Ratones , Estrés OxidativoRESUMEN
BACKGROUND: Cryptorchidism and oligozoospermia are clinical conditions closely associated with impaired fertility. Oxidative stress and related sperm DNA damage have been identified as significant causes of male infertility. AIM: To determine the extent of sperm nuclear DNA damage in patients affected with idiopathic oligozoospermia or undescended testes and to examine its relationship with oxidative stress. PATIENTS AND METHODS: We studied 20 patients with idiopathic oligozoospermia and 18 with undescended testes (who previously underwent orchiopexy) and 25 normozoospermic healthy controls. All subjects underwent semen analysis. Sperm DNA damage was evaluated by the sperm chromatin structure assay/flow cytometry (SCSA-FCM) and by the dUTP-biotin nick end labeling (TUNEL) assay. Levels of reactive oxygen species (ROS) and total antioxidant capacity (TAC) were assessed by a chemiluminescence assay. RESULTS: DFI (percentage of sperm with denatured DNA) values and percentage of TUNEL positive cells were significantly greater in patients with oligozoospermia (DFI: 28.8+/-5.6; TUNEL+: 26.9+/-3.0) or cryptorchidism (DFI: 26.4+/-10.1; TUNEL+: 29.1+/-3.9), compared with controls (DFI: 7.1+/-0.9; TUNEL+: 14.2+/-1.2). Similarly, both groups of patients had significantly higher (p<0.01) levels of ROS. TAC levels did not differ between control and patient groups, suggesting that the DNA damage occurs before spermiation. CONCLUSIONS: Sperm DNA damage is significantly increased in men with idiopathic oligozoospermia and in cryptorchid subjects. The finding of increased ROS levels may indicate that seminal oxidative stress may be involved in the pathogenesis of sperm DNA damage in these patients.
Asunto(s)
Cromatina/genética , Daño del ADN , Infertilidad Masculina/genética , Estrés Oxidativo , Espermatozoides , Adulto , Estudios de Casos y Controles , Criptorquidismo/complicaciones , Criptorquidismo/genética , Fragmentación del ADN , Citometría de Flujo , Humanos , Etiquetado Corte-Fin in Situ , Infertilidad Masculina/fisiopatología , Masculino , Persona de Mediana Edad , Oligospermia/complicaciones , Oligospermia/genética , Especies Reactivas de Oxígeno/análisis , Índice de Severidad de la Enfermedad , Estadísticas no ParamétricasRESUMEN
OBJECTIVE: To investigate the expression of Fas protein on the surface of ejaculated spermatozoa of normozoospermic and nonnormozoospermic men. DESIGN: Prospective study. SETTING: University infertility clinic. PATIENT(S): Twenty-three volunteer normozoospermic men (controls) and 43 men undergoing infertility evaluation (cases). INTERVENTION(S): Analysis of ejaculated spermatozoa by indirect immunofluorescence of Fas protein by flow cytometry. MAIN OUTCOME MEASURE(S): Comparison of flow cytometric analysis of autofluorescence, control tests (secondary antibody and isotype control), and experimental tests (anti-Fas monoclonal antibody) in the spermatozoa of ejaculated samples. RESULT(S): No expression of Fas protein was found on the surface of ejaculated spermatozoa of controls and cases. CONCLUSION(S): The Fas molecules are not present in substantial amounts in the ejaculated spermatozoa of normozoospermic and nonnormozoospermic men. Therefore, our results do not support the "abortive apoptosis" theory.
Asunto(s)
Citometría de Flujo , Infertilidad Masculina/metabolismo , Espermatozoides/química , Donantes de Tejidos , Receptor fas/análisis , Anticuerpos Monoclonales , Estudios de Casos y Controles , Eyaculación , Fluorescencia , Técnica del Anticuerpo Fluorescente Indirecta , Humanos , MasculinoRESUMEN
Embryo development depends on maternal and embryonic factors that may regulate genetic programs in early development. Effects of growth factors on proliferation, differentiation and morphogenesis along embryogenesis have been documented. However, studies have not established the role of growth factors in the preimplantational period. The purpose of this study was to investigate the possible effects of growth factors and embryo density on mouse preimplantation development in vitro. Two-and eight-cell CF-1 embryos were cultured individually or in groups of ten in HTF medium, alone or with EGF, TGF-beta1 and IGF-I. Cleavage rate varied greatly with growth factors and increased significantly when eight-cell embryos were cultured in groups. On the other hand, when two-cell embryos were cultured in group, the cleavage rate was slower than that obtained when embryos were individually cultured. The differentiation rate increased significantly in two-cell embryos cultured in groups (p<0.05). EGF, TGF-beta1 and IGF-I increased differentiation rates significantly in two-cell embryos individually cultured for 68 hours. The combination of EGF and TGF-beta1 increased the differentiation rates significantly. The other combinations were not effective in modifying this parameter. Hatching rates increased in embryos cultured in groups (p<0.05). TGF-beta1 decreased this parameter significantly in two-or eight-cell embryos cultured in groups (p<0.05). The data described in this report suggest that preimplantational mouse embryos produce some factor or factors that enhance its development, specially the differentiation and hatching rates. However, a functional role for polypeptide growth factors during preimplantational development has to be determined. (AU)
Asunto(s)
Técnicas In Vitro , Ratones , Animales , Desarrollo Fetal/efectos de los fármacos , Semillas , Blastocisto , Estructuras Embrionarias/crecimiento & desarrollo , Sustancias de Crecimiento/farmacología , Ratones EndogámicosRESUMEN
Embryo development depends on maternal and embryonic factors that may regulate genetic programs in early development. Effects of growth factors on proliferation, differentiation and morphogenesis along embryogenesis have been documented. However, studies have not established the role of growth factors in the preimplantational period. The purpose of this study was to investigate the possible effects of growth factors and embryo density on mouse preimplantation development in vitro. Two-and eight-cell CF-1 embryos were cultured individually or in groups of ten in HTF medium, alone or with EGF, TGF-beta1 and IGF-I. Cleavage rate varied greatly with growth factors and increased significantly when eight-cell embryos were cultured in groups. On the other hand, when two-cell embryos were cultured in group, the cleavage rate was slower than that obtained when embryos were individually cultured. The differentiation rate increased significantly in two-cell embryos cultured in groups (p<0.05). EGF, TGF-beta1 and IGF-I increased differentiation rates significantly in two-cell embryos individually cultured for 68 hours. The combination of EGF and TGF-beta1 increased the differentiation rates significantly. The other combinations were not effective in modifying this parameter. Hatching rates increased in embryos cultured in groups (p<0.05). TGF-beta1 decreased this parameter significantly in two-or eight-cell embryos cultured in groups (p<0.05). The data described in this report suggest that preimplantational mouse embryos produce some factor or factors that enhance its development, specially the differentiation and hatching rates. However, a functional role for polypeptide growth factors during preimplantational development has to be determined.