Centro ANDROGEN

A Coruña, Spain

Centro ANDROGEN

A Coruña, Spain
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Esteves S.C.,Androfert andrology and Human Reproduction Clinic | Zini A.,McGill University | Aziz N.,Liverpool Womens Hospital | Alvarez J.G.,Centro ANDROGEN | And 2 more authors.
Urology | Year: 2012

In 2010, the World Health Organization established new reference values for human semen characteristics that are markedly lower than those previously reported. Despite using controlled studies involving couples with a known time to pregnancy to establish the new limits, the reference studies are limited with regard to the population analyzed and the methods used for semen evaluation. The present review discusses concerns related to the new reference values for semen characteristics, including the effect on patient referral, diagnosis, and treatment of recognized conditions, such as varicocele, and on the indications for assisted reproductive technologies. © 2012 Elsevier Inc.


Buffone M.G.,CONICET | Calamera J.C.,Laboratorio Of Estudios En Reproduccion Ler | Brugo-Olmedo S.,Centro Medico Seremas | De Vincentiis S.,Centro Medico Seremas | And 5 more authors.
Fertility and Sterility | Year: 2012

Objective: To investigate the correlation between sperm superoxide dismutase (SOD) content and motility recovery after thawing of cryopreserved human sperm, based on the rationale that this antioxidant enzyme provides protection against reactive oxygen species-induced damage during cryopreservation. Design: Prospective study. Setting: Private infertility institute and university-based research laboratory. Patient(s): Forty-two consenting normozoospermic patients consulting for infertility. Intervention(s): The SOD content was measured in sperm from unfractionated samples and in sperm recovered from the pellet fraction obtained after discontinuous density gradient centrifugation. Main Outcome Measure(s): Sperm motility was evaluated post-thaw in the two sets of samples and motility recovery was plotted against the sperm SOD content to determine their correlation. Result(s): There was a significant positive correlation between motility recovery after thawing and SOD content in sperm from the 90% gradient pellet containing highly purified mature sperm. There was also a significant negative correlation between motility after thawing and SOD content in the unfractionated sample. Conclusion(s): The positive correlation between post-thaw motility recovery and SOD content in mature spermatozoa provides a good predictor of post-thaw motility recovery after cryopreservation. © 2012 American Society for Reproductive Medicine, Published by Elsevier Inc.


Kocer A.,French Institute of Health and Medical Research | Henry-Berger J.,French Institute of Health and Medical Research | Noblanc A.,French Institute of Health and Medical Research | Champroux A.,French Institute of Health and Medical Research | And 13 more authors.
Free Radical Biology and Medicine | Year: 2015

Normal embryo and foetal development as well as the health of the progeny are mostly dependent on gamete nuclear integrity. In the present study, in order to characterize more precisely oxidative DNA damage in mouse sperm we used two mouse models that display high levels of sperm oxidative DNA damage, a common alteration encountered both in in vivo and in vitro reproduction. Immunoprecipitation of oxidized sperm DNA coupled to deep sequencing showed that mouse chromosomes may be largely affected by oxidative alterations. We show that the vulnerability of chromosomes to oxidative attack inversely correlated with their size and was not linked to their GC richness. It was neither correlated with the chromosome content in persisting nucleosomes nor associated with methylated sequences. A strong correlation was found between oxidized sequences and sequences rich in short interspersed repeat elements (SINEs). Chromosome position in the sperm nucleus as revealed by fluorescent in situ hybridization appears to be a confounder. These data map for the first time fragile mouse sperm chromosomal regions when facing oxidative damage that may challenge the repair mechanisms of the oocyte post-fertilization. © 2015 Elsevier Inc. All rights reserved.


Cabrillana M.E.,National University of Cuyo | Cabrillana M.E.,University of Aconcagua | Uribe P.,University of the Frontier | Villegas J.V.,University of the Frontier | And 5 more authors.
Systems Biology in Reproductive Medicine | Year: 2016

Peroxynitrite is a highly reactive nitrogen species and when it is generated at high levels it causes nitrosative stress, an important cause of impaired sperm function. High levels of peroxynitrite have been shown to correlate with decreased semen quality in infertile men. Thiol groups in sperm are mainly found in enzymes, antioxidant molecules, and structural proteins in the axoneme. Peroxynitrite primarily reacts with thiol groups of cysteine-containing proteins. Although it is well known that peroxynitrite oxidizes sulfhydryl groups in sperm, the subcellular localization of this oxidation remains unknown. The main objective of this study was to establish the subcellular localization of peroxynitrite-induced nitrosative stress in thiol groups and its relation to sperm motility in human spermatozoa. For this purpose, spermatozoa from healthy donors were exposed in vitro to 3-morpholinosydnonimine (SIN-1), a compound which generates peroxynitrite. In order to detect peroxynitrite and reduced thiol groups, the fluorescent probes, dihydrorhodamine 123 and monobromobimane (mBBr), were used respectively. Sperm viability was analyzed by propidium iodide staining. Peroxynitrite generation and thiol redox state were monitored by confocal microscopy whereas sperm viability was evaluated by flow cytometry. Sperm motility was analyzed by CASA using the ISAS® system. The results showed that exposure of human spermatozoa to peroxynitrite results in increased thiol oxidation which is mainly localized in the sperm head and principal piece regions. Thiol oxidation was associated with motility loss. The high susceptibility of thiol groups to peroxynitrite-induced oxidation could explain, at least in part, the negative effect of reactive nitrogen species on sperm motility. Abbreviations: DHR: dihydrorhodamine 123; mBBr: monobromobimane ONOO−: peroxynitrite RNS: reactive nitrogen species RFI: relative fluorescence intensity SIN-1: 3-morpholinosydnonimine CASA: Computer-Aided Sperm Analysis PARP: poli ADP ribose polimerasa VCL: curvilinear velocity VSL: straight-line velocity VAP: average path velocity PRDXs: peroxiredoxins ODF: outer dense fiber ODF1: outer dense fiber 1 PI: propidium iodide DMSO: dimethyl sulfoxide SD: standard deviation ANOVA: analysis of variance © 2016 Taylor & Francis


Cheuqueman C.,University of the Frontier | Loren P.,University of the Frontier | Arias M.,University of the Frontier | Risopatron J.,University of the Frontier | And 4 more authors.
Theriogenology | Year: 2015

Stress tolerance can be induced in embryos by oocyte exposure to hydrostatic pressure, osmotic agents, heat shock, or reactive oxygen species. This study assessed the effects of exposing bovine oocytes to a nitric oxide (NO) donor, sodium nitroprusside (SNP), on subsequent invitro embryo production, embryo quality and the expression of genes involved in NO production (iNOS, eNOS, and nNOS), stress tolerance (HSP70 and HSP90), oxidative stress (HIF1A and PRDX5), and apoptosis (BCL2A1). Invitro mature oocytes were incubated with SNP (control, 10-6M, 10-5M, and 10-4M) for 1hour before invitro fertilization, and cultured until Day 7. Cleavage and blastocyst rates were recorded. Next, embryo quality (ratio of inner cell mass to total cell number) and relative gene expression of iNOS, eNOS, nNOS, HSP70, HSP90, HIF1A, PRDX5, and BCL2A1 were determined in expanded blastocysts. Cleavage rates were significantly lower for 10-4M SNP compared with the control and 10-5M SNP treatments (77±7.1%, 82±8.4%, and 84.9±4.1%, respectively). Total blastocyst rates were lower in the 10-4M SNP group relative to the control group (26.2±4.9% and 34.1±7.8%, respectively). Embryo quality was similar among the groups. However, our relative gene expression analysis revealed the downregulation of endothelial oxide nitric synthase messenger RNA in expanded blastocysts in all the treatment groups compared with the control treatment. These results suggest that the short-term exposure of mature bovine oocytes to a NO donor does not induce their stress tolerance and has no beneficial effect on the invitro embryo production of bovine embryos. © 2015 Elsevier Inc.


Cheuqueman C.,University of the Frontier | Arias M.E.,University of the Frontier | Risopatron J.,University of the Frontier | Felmer R.,University of the Frontier | And 3 more authors.
Andrologia | Year: 2015

Gamete co-incubation generates high free radical levels surrounding growing zygotes which may impair subsequent embryo viability. Melatonin eliminates a wide variety of free radicals; hence, we tried to improve in vitro embryo production by adding melatonin to in vitro fertilisation (IVF) media in high (Exp. 1) and low concentrations (Exp. 2), and we evaluated its effect on bull sperm function during IVF co-incubation time (Exp. 3). In Experiment 1, we supplemented IVF media culture with 0.01, 0.1 and 1 mmol of melatonin, along with a no melatonin control group. In Experiment 2, melatonin levels were reduced to 10, 100 and 1000 nmol, with a no melatonin control group. In Experiment 3, spermatozoa were incubated in IVF media with melatonin (as Exp. 2) and functional parameters were analysed at 0, 4 and 18 h. In Experiment 1, only 1 mmol melatonin showed lesser blastocyst rates than control (C: 23.2 ± 6.7% versus 1 mmol: 2.0 ± 1.7%). In Experiment 2, no statistical differences were found in cleavage percentage, blastocyst percentage and total cell count for any melatonin treatment. In Experiment 3, sperm samples with 1000 nmol melatonin had a significantly higher wobbler (WOB) coefficient, a lower percentage of intact acrosomes, a lower percentage of viable spermatozoa with ROS, greater DNA fragmentation and higher DNA oxidation than controls. Total fluorescence intensity for ROS at 10 nmol melatonin was significantly greater than controls (P < 0.05). IVF media with 1 mmol melatonin is deleterious for embryo development, and in lower concentrations, it modulated sperm functionality, but had no effects on embryo production. © 2014 Blackwell Verlag GmbH.


PubMed | University of the Frontier, Autonomous University of Barcelona and Centro ANDROGEN
Type: | Journal: Andrologia | Year: 2016

Retrospective analysis of monthly embryo production from December 2011 to May 2015 and its correlation with meteorological data in our geographic zone was made. We had observed that in certain time of the year, in vitro blastocyst production decreases. Accordingly, was examined the association between blastocyst production and climatological parameters. Cleavage rates correlate positively with blastocyst rates (p<.05). Significant differences in cleavage rates between autumn and summer (79.8%; 71.5%), and between winter and autumn (71.8%; 79.8%), were found. Blastocyst production had lower efficiency in June (912%) and July (4.95.7%), which coincides with winter season. In contrast, higher embryo production was obtained in February (22.29.7%), March (22.914%) and September (25.26.6%), which coincides with autumn and spring season. Similarly, embryo production correlates with meteorological parameters: blastocyst production positively correlates with sunshine hours, maximum temperature and average temperature. Similarly, blastocyst production inversely correlates with total precipitation and days >1mm precipitation (p<.05). There is a significant decrease in bovine in vitro embryo production efficiency during winter season in our warm-summer Mediterranean climate zone. It remains to be investigated the direct effect of environmental factors on oocyte quality and its impact on in vitro production efficiency.


PubMed | University of the Frontier, Autonomous University of Barcelona and Centro ANDROGEN
Type: Journal Article | Journal: Theriogenology | Year: 2015

Stress tolerance can be induced in embryos by oocyte exposure to hydrostatic pressure, osmotic agents, heat shock, or reactive oxygen species. This study assessed the effects of exposing bovine oocytes to a nitric oxide (NO) donor, sodium nitroprusside (SNP), on subsequent in vitro embryo production, embryo quality and the expression of genes involved in NO production (iNOS, eNOS, and nNOS), stress tolerance (HSP70 and HSP90), oxidative stress (HIF1A and PRDX5), and apoptosis (BCL2A1). In vitro mature oocytes were incubated with SNP (control, 10(-6) M, 10(-5) M, and 10(-4) M) for 1 hour before in vitro fertilization, and cultured until Day 7. Cleavage and blastocyst rates were recorded. Next, embryo quality (ratio of inner cell mass to total cell number) and relative gene expression of iNOS, eNOS, nNOS, HSP70, HSP90, HIF1A, PRDX5, and BCL2A1 were determined in expanded blastocysts. Cleavage rates were significantly lower for 10(-4) M SNP compared with the control and 10(-5) M SNP treatments (77 7.1%, 82 8.4%, and 84.9 4.1%, respectively). Total blastocyst rates were lower in the 10(-4) M SNP group relative to the control group (26.2 4.9% and 34.1 7.8%, respectively). Embryo quality was similar among the groups. However, our relative gene expression analysis revealed the downregulation of endothelial oxide nitric synthase messenger RNA in expanded blastocysts in all the treatment groups compared with the control treatment. These results suggest that the short-term exposure of mature bovine oocytes to a NO donor does not induce their stress tolerance and has no beneficial effect on the in vitro embryo production of bovine embryos.


Alvarez J.G.,Centro Androgen | Alvarez J.G.,Harvard University
Revista Internacional de Andrologia | Year: 2011

Introduction and objectives: Previous studies have shown that the membrane and desoxiribonucleic acid (DNA) damage observed in ejaculated spermatozoa is mostly produced post-testicularly, during passage of sperm through the epididymis. This damage is induced, for the most part, by free radicals and amplified by proinflammatory factors. The objective of this study was to evaluate the efficacy and safety of docosahexaenoic acid (DHA), a longchain polyunsaturated fatty acid with anti-inflammatory and antioxidant properties, in the treatment of male infertility associated to sperm membrane and DNA damage. Material and methods: A total of 80 patients diagnosed of necrozoospermia, measured as sperm viability, or of an abnormal sperm DNA fragmentation index (DFI), measured by the TUNEL test, were included in the study. In the necrozoospermia group, as well as in the abnormal DNA fragmentation (DFI >20%) group, 20 patients were included in the experimental group treated with 200mg of DHA derived from the microalgae Schizochytrium sp. (GestaDha ®) (patent by Martek Bioscience Corporation under registered trademark life's DHA™) every 12 hours, and 20 patients were included in the control group not treated. The duration of the treatment was of 8 weeks and semen samples were obtained at 0, 1, 2, 4 y 8 weeks of treatment for the analysis of sperm viability and the DFI. Results: The data showed that GestaDha ® significantly decreased both the rate of necrozoospermia and the DFI after the first week of treatment compared to the control group. Differences between the experimental and control groups were statistically significant for both parameters (p< 0.01). No adverse effects were observed in any of the patients treated with GestaDha ®. Conclusions: This study shows, for the first time, the efficacy and safety of GestaDha ® in the treatment of male infertility associated to sperm membrane and DNA damage. © 2011 Sociedad Española de Andrología.


Noblanc A.,French Institute of Health and Medical Research | Damon-Soubeyrand C.,French Institute of Health and Medical Research | Karrich B.,French Institute of Health and Medical Research | Henry-Berger J.,French Institute of Health and Medical Research | And 11 more authors.
Free Radical Biology and Medicine | Year: 2013

Gamete DNA integrity is one key parameter conditioning reproductive success as well as the quality of life for the offspring. In particular, damage to the male nucleus can have profound negative effects on the outcome of fertilization. Because of the absence of repair activity of the quiescent mature spermatozoa it is easily subjected to nuclear damage, of which oxidative damage is by far the most prominent. In relation to the organization of the mammalian sperm nucleus we show here that one can correlate the nuclear regions of lower compaction with areas preferentially showing oxidative damage. More precisely, we show that oxidative DNA damage targets primarily histone-rich and nuclear matrix-attached domains located in the peripheral and basal regions of the mouse sperm nucleus. These particular sperm DNA domains were recently shown to be enriched in genes of paramount importance in postfertilization DNA replication events and in the onset of the embryonic developmental program. We propose that monitoring of sperm DNA oxidation using the type of assay presented here should be considered in clinical practice when one wants to estimate the integrity of the paternal nucleus along with more classical assays that essentially analyze DNA fragmentation and nucleus compaction. © 2013 Elsevier Inc.

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