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Biomedical subjects

Nicolás Garrido

Publications and source records attributed to Nicolás Garrido.

At least 19 recordsLinked to original sources
PubMed2006

The effectiveness of modified sperm washes in severely oligoasthenozoospermic men infected with human immunodeficiency and hepatitis C viruses.

Men who are seropositive for human immunodeficiency virus (HIV) or hepatitis C virus now can father children safely, thanks to sperm washing and nested polymerase chain-reaction techniques, followed by intracytoplasmic sperm injection. However, in a small percentage of such patients, it is impossible to recover spermatozoa after said procedures because of their highly impaired spermatogenesis. We have established that less rigorous methods, such as repeated centrifugation, yield nested polymerase chain-reaction HIV- and hepatitis C virus-negative specimens, even in sperm samples from men with severe oligoasthenozoospermia.

Adult↗
PubMed2006

Effect of sperm glutathione peroxidases 1 and 4 on embryo asymmetry and blastocyst quality in oocyte donation cycles.

OBJECTIVE: To prospectively determine the impact of concrete components of the sperm oxidative glutathione stress system in terms of enzymatic activity and mitochondrial RNA (mRNA) expression on embryo quality and reproductive outcome. Human spermatozoa use the glutathione system to inactivate reactive oxygen metabolites, and there is a close correlation between some components of the glutathione system and male fertility. However, very few data are published regarding this system in sperm cells and its effect on fertilization ability and embryo development in human beings. DESIGN: An oocyte-donation model, used to homogenize the female factor. SETTING: University-affiliated private IVF setting. PATIENT(S): Semen samples from infertile males (n = 43) of couples undergoing oocyte-donation cycles (n = 43). INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Gene expression and activity of glutathione peroxidases (GPXs) 1 and 4, glutathione reductase, and intracellular glutathione (GSH) by fluorescent quantitative polymerase chain reaction and spectrophotometry, respectively. RESULT(S): Fertilization rate, pronuclear number, asymmetry, and pronuclear body distribution were not correlated with any sperm glutathione parameters that were considered. When day 3 embryo parameters were evaluated, only GPX4 mRNA expression in sperm cells was statistically significantly lower when asymmetric embryos were observed. Also, worst embryo development and morphology on day 5 was statistically significantly correlated with lower sperm GPX1 activity (101.07 vs. 258.8 IU/mg protein). Glutathione system analysis in fresh sperm was not statistically significantly different in patients achieving pregnancy compared with those who not, and we did not find any correlation with implantation rate. CONCLUSION(S): We have been able to correlate embryo morphology on day 3 with the sperm expression of GPX family members. The results indicate that sperm-derived mRNA may condition human embryo quality and persist even to blastocyst stage. The correlation of the sperm GPX family mRNA expression with embryo health appears quite promising for discovery of molecular causes of male infertility.

Adult↗
PubMed2006

Value of the sperm deoxyribonucleic acid fragmentation level, as measured by the sperm chromatin dispersion test, in the outcome of in vitro fertilization and intracytoplasmic sperm injection.

OBJECTIVE: To determine the prognostic value of sperm DNA fragmentation levels, as measured by the sperm chromatin dispersion (SCD) test, in predicting IVF and ICSI outcome. DESIGN: Double-blind prospective study. SETTING: University-affiliated private IVF setting. PATIENT(S): A total of 85 couples undergoing infertility treatment with IVF/ICSI. INTERVENTION(S): Analysis of DNA fragmentation by the SCD test in 170 aliquots obtained from the ejaculate and from the processed semen used for assisted reproductive technologies (ART). MAIN OUTCOME MEASURE(S): Percentage of spermatozoa with fragmented DNA was statistically correlated with embryo quality and reproductive success. RESULT(S): Fertilization rate was inversely correlated with DNA fragmentation (r = -0.245 P = .045). Higher DNA fragmentation rate gave an increased proportion of zygotes showing asynchrony between the nucleolar precursor bodies of zygote pronuclei (73.8% vs. 28.8% P < .001). In addition, the slower embryo development and worst morphology on day 6 was correlated with higher sperm DNA fragmentation (47.7% vs. 29.4% P = .044). We also observed a negative correlation between DNA fragmentation and the implantation rate (r = -0.250 P = .042). However, SCD test values were not statistically different in cycles that resulted in a pregnancy compared with those that did not (33.2 vs. 28.2 and 32.4 vs. 34.7). CONCLUSION(S): This is the first report that describes a correlation between sperm DNA integrity, as measured by the SCD test, and fertilization rate, embryo quality, and implantation rate in IVF/ICSI. The degree of DNA fragmentation was inversely correlated with fertilization rate, synchrony of the nucleolar precursor bodies' pattern in pronuclei, embryo ability to achieve blastocyst stage, and embryo morphological quality. Because SCD test values were correlated with embryo quality and blastocyst rate, the lack of correlation between sperm DNA fragmentation and pregnancy outcome in IVF might be due to embryo selection before transfer. The ability of the SCD test to predict the blastocyst rate after IVF/ICSI warrants further study.

Adult↗
PubMed2006

Sperm cryopreservation in oncological patients: a 14-year follow-up study.

OBJECTIVE: Oncologic treatments can destroy spermatogenic dividing cells and cause azoospermia which could be irreversible. Sperm banking is the best option to preserve male fertility after these treatments. It is easy, inexpensive, and safe. To date, few clinical data are available about large series of cancer patients. Our objective was to determine the usefulness of these preventive sperm freezing protocols. DESIGN: Prospective study. SETTING: University-affiliated private fertility center. PATIENT(S): One hundred eighty-six cancer patients who banked sperm samples at our center before surgery or chemo- or radiotherapy treatments from 1991 to 2004. INTERVENTION(S): Conjugal status, age, type of cancer, treatment, and future use (if any) of the cryopreserved sperm samples for assisted reproduction technology (ART), and cycle results were recorded, analyzed, and compared with a control group. MAIN OUTCOME MEASURE(S): Basic sperm analysis of semen samples from cancer patients prior to freezing, after thawing, and after capacitation for ART. RESULT(S): A total of 320 semen samples were frozen before antineoplasic treatment. Six months later, 27% of the males recovered normal sperm production. From all frozen samples, 8.7% were discarded; the reasons were pregnancy achievement (55%), normal sperm production (28%), and patient death (18%). Finally, 5 IUI cycles and 30 ICSI cycles were done from frozen samples, with 1 and 15 pregnancies, respectively; results were comparable with those obtained in a control group. CONCLUSION(S): A significant number of males who cryopreserved semen samples before receiving antitumoral treatments have employed them. The results obtained showed that this is the strategy of choice, aiming to preserve fertility for the future, because the cost/benefit ratio is favorable. Patients should be counseled accordingly.

Adult↗
PubMed2005

Value of the sperm chromatin dispersion test in predicting pregnancy outcome in intrauterine insemination: a blind prospective study.

BACKGROUND: Sperm DNA integrity has been used as a new marker of sperm quality in the prediction of pregnancy. Nevertheless, no previous study has been performed by analysing the same samples that were employed in assisted reproduction. The main objective of this work was to correlate sperm chromatin dispersion (SCD), measured by the SCD test, with semen parameters and pregnancy outcome in intrauterine insemination (IUI). METHODS: A total of 100 semen samples obtained from males of couples undergoing IUI were analysed by the SCD test before and after swim-up, and the results were correlated with semen parameters and pregnancy outcome. RESULTS: SCD was negatively correlated with sperm motility in both ejaculated and processed semen. Sperm recovered by swim-up did not show a significant improvement in DNA integrity. No correlation was found between SCD and pregnancy outcome in IUI. CONCLUSIONS: DNA dispersion, as measured by the SCD test, is not correlated with pregnancy outcome in IUI.

Chromatin↗
PubMed2005

How would revealing the identity of gamete donors affect current practice?

The possibility of revealing the identity of gamete donors may change the decision of a donor to collaborate with infertility clinics and patients. Previous experiences with patients and donors, as well as different cultural viewpoints in northern and southern Europe, are presented.

Confidentiality↗
PubMed2004

Semen characteristics in human immunodeficiency virus (HIV)- and hepatitis C (HCV)-seropositive males: predictors of the success of viral removal after sperm washing.

BACKGROUND: Human immunodeficiency virus (HIV)/hepatitis C virus (HCV)-seropositive males can now father children safely, avoiding transmission risks to the mother and the children using sperm washing and nested PCR (nPCR) techniques. Nevertheless, we still lack enough data to determine the reasons why approximately 10% of the performed sperm washes remain positive, thus forcing the repetition of the treatment. Semen quality in infected males is also essential to these procedures. We aimed to determine the predictive value of the semen parameters, sperm washing procedure and the infection status for the post-wash viral positivity, as well as the correlation between the semen and the disease features. METHODS: Semen characteristics were evaluated in 136 samples provided from 125 males. We also included a control group of 125 males matched by age and length of sexual abstinence. At the time of semen retrieval, 70 of them were infected with HIV (45 also with HCV, 64.3%), and 55 of them with HCV alone. nPCR for viral detection was performed in each sample. RESULTS: Thirteen out of 136 (9.5%) of the samples were positive for one or more viral detections (HIV RNA, HIV DNA and HCV RNA, when needed). From a total of 240 nPCR viral analyses, 16 were positive (6.6%). None of the seminal parameters were adequate to predict post-wash results, nor was a positive result dependent on the volumes used in the semen wash. A positive correlation was found between post-wash progressive motility and CD4 blood levels, as well as a negative correlation between progressive motility and time of evolution of the disease in HIV-infected males. CONCLUSIONS: Semen analysis, according to the World Health Organization criteria, of HIV- and HCV-affected patients showed no differences from that of non-infected males. Moreover, low CD4 blood levels, and a long evolution of the disease do not negatively affect sperm motility.

Anti-Retroviral Agents↗
PubMed2004

Report of the results of a 2 year programme of sperm wash and ICSI treatment for human immunodeficiency virus and hepatitis C virus serodiscordant couples.

BACKGROUND: Human immunodeficiency virus (HIV) and hepatitis C virus (HCV) serodiscordant couples with the male infected can be helped to have children minimizing the transmission risk. Our aim was to evaluate the results of our assisted reproduction programme for these couples and to develop adequate strategies for their management. METHODS: Members of serodiscordant couples: (i) HIV positive males attending our centre for sperm wash and assisted reproduction and (ii) HCV positive males needing assisted reproduction for infertility provided 134 semen samples for sperm wash. Before ICSI treatment, semen was confirmed to be negative for viral presence by reverse transcription and nested PCR after the sperm wash. RESULTS: Sperm washes were effective in 90% of the samples. Regardless of the type of infection, no differences were found in semen quality, embryos obtained and pregnancy rates (40-48% per cycle). To date, 41 pregnancies and 23 newborns were obtained. Fertilization rates were lower in HCV than in HIV serodiscordant couples (59.3+/-5.3% versus 72.0+/-8.1%), probably because they were infertile couples for whom we recommended sperm wash and PCR. No seroconversion was detected in the patients' follow-up. CONCLUSIONS: To date, sperm wash, nested PCR and ICSI is a safe and effective procedure that avoids HIV and HCV transmission with reasonable pregnancy rates, and is cost-effective.

Adult↗
PubMed2004

Relationship between standard semen parameters, calcium, cholesterol contents, and mitochondrial activity in ejaculated spermatozoa from fertile and infertile males.

PURPOSE: To correlate levels of cholesterol (CH), calcium (Ca2+), and mitochondrial activity (MA) with the standard semen parameters and to compare them between fertile and infertile men. METHODS: We studied 151 semen samples from infertile (n = 60) or fertile (n = 91) males. Basic sperm parameters were analyzed. Ca2+ and CH concentrations on seminal plasma were determined by enzymoimmunoanalysis. Intracellular Ca2+ and CH concentrations in the sperm plasma membrane and mitochondrial activity by fluorometry. RESULTS: There was a significant positive correlation between sperm membrane CH and sperm morphology. Intracellular Ca2+ was lower in infertile patients compared to fertile. No differences were found regarding Ca2+ and CH concentrations in seminal plasma. MA is directly and strongly related with sperm motility. CONCLUSIONS: Intracellular concentrations of Ca2+ and the proportion of CH in the sperm membrane are two important markers of the sperm quality due to its direct relationship with sperm morphology and fertility potential.

Calcium↗
PubMed2004

Role of cholesterol, calcium, and mitochondrial activity in the susceptibility for cryodamage after a cycle of freezing and thawing.

OBJECTIVE: To correlate the levels of two concrete sperm markers, cholesterol and Ca(+2), together with mitochondrial activity on raw semen samples with the post-thaw recovery of spermatozoa with progressive motility on human sperm samples as the first step to improve sperm cryostorage protocols. DESIGN: Controlled prospective research project. SETTING: Private and university-affiliated setting. PATIENT(S): Semen samples from 122 males attending our center for infertility (n = 47) or semen donation (n = 75) were studied. INTERVENTION(S): The mean basic semen parameters of the 122 semen samples studied before and after the freezing and thawing process. MAIN OUTCOME MEASURE(S): We determined Ca(+2) and cholesterol concentrations on seminal plasma by enzymoimmunoanalysis techniques, intracellular Ca(+2) concentrations, cholesterol concentrations in the sperm plasma membrane and mitochondrial activity by fluorometry. RESULT(S): Cholesterol concentration in seminal plasma and cholesterol contents in the sperm membrane and mitochondrial activity were studied. No correlations were initially found to be of statistical significance. Regarding seminal plasma and intracellular sperm Ca(+2) concentrations, a statistically significant negative correlation was found (P=.036 and P=.016). CONCLUSION(S): Higher cholesterol contents do not appear to protect sperm against cryodamage. Conversely, Ca(+2) equilibrium appears to be essential for a good post-thaw recovery. Mitochondrial activity is not reflecting the possibilities of sperm survival and is probably not a good indicator of the sperm metabolism.

Calcium↗
PubMed2004

Relationship among standard semen parameters, glutathione peroxidase/glutathione reductase activity, and mRNA expression and reduced glutathione content in ejaculated spermatozoa from fertile and infertile men.

OBJECTIVE: To determine the expression and enzymatic activity of glutathione peroxidase (GPX)-1, GPX-4, and glutathione reductase together with glutathione (GSH) concentrations in spermatozoa from fertile and infertile men. DESIGN: Prospective study. SETTING: University-affiliated private center. PATIENT(S): Fifty-four infertile men undergoing assisted reproduction techniques and 55 fertile sperm donors with pregnancies and newborns by artificial insemination. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Analysis of gene expression by fluorescent quantitative polymerase chain reaction and an analysis of enzymatic activity and GSH concentration by controlled biochemical reactions and spectrophotometry. RESULT(S): GPX-4 activity but not mRNA expression is directly related to sperm morphology (strict criteria) and is more compromised with a low percentage of normal sperm. These differences are also demonstrated when fertile and infertile men were compared. In addition, intracellular GSH concentrations are lower when sperm morphology is severely impaired, but no differences were found between fertile and infertile men. CONCLUSION(S): Intracellular sperm GSH system components GPX-4 and GSH are altered in infertile men, and these alterations seem to be linked to sperm morphology.

Case-Control Studies↗
PubMed2004

Testicular sperm extraction (TESE) and intracytoplasmic sperm injection (ICSI) in hypogonadotropic hypogonadism with persistent azoospermia after hormonal therapy.

PURPOSE: We aimed to retrieve testicular sperm to be employed on intracytoplasmic sperm injection (ICSI) cycles on a male affected of hypogonadotropic hypogonadism (HH) that remained azoospermic after long-time hormonal treatment. METHODS: Design. We initially performed hormonal therapy using gonadotropins to achieve spermatogenesis. After several semen analyses, we weighed the possibility of looking for testicular spermatozoa for ICSI. Setting. A private university-affiliated setting. Patient. A 30-years-old man diagnosed 10 years ago to suffer from idiopathic, prepubertal HH. Interventions. Gonadotrophin treatment was initiated with hCG and follicle stimulating hormone (FSH). Testicular sperm extraction was carried out when repeated spermiograms were negative. Motile testicular spermatozoa were cryopreserved and were subsequently employed for ICSI. Multiple follicular development was stimulated with gonadotropins after a downregulation with gonadotropin-releasing hormone (GnRH) antagonists in the woman. Main Outcome Measures. Seminal analyses were performed after 3, 6, and 12 months of treatment and serum FSH, luteinizing hormone (LH) and testosterone levels were also measured. RESULTS: Seminal analysis showed always azoospermia. Serum FSH was 2.9 mIU/mL, serum LH >1 mIU/mL and serum testosterone 7.9 ng/mL (12 months after treatment). Nine oocytes were collected by ultrasound-guided transvaginal route and eight of them were microinjected with motile, frozen-thawed testicular spermatozoa. Four oocytes were fertilized. Three embryos were transferred without pregnancy. CONCLUSIONS: The case report here presented shows that the currently available assisted reproduction techniques may be of value in patients with HH not responding to conventional hormonal treatments.

Adult↗
PubMed2004

Pro-oxidative and anti-oxidative imbalance in human semen and its relation with male fertility.

Oxidative stress is a common condition suffered by biological systems in aerobic conditions. Human semen also has its own molecular guard against the free radicals created by normal respiratory process or from immune reactions. The equilibrium of the creation and scavenging of free radicals is mandatory in the spermatozoa to fertilize and initiate a full-term pregnancy. The paper is a systematic review of publications that evaluate oxidative stress in semen. The Cochrane Library, Medline (1966-2003), Embase (1988-2003), SciSearch (1981-2003) and the conference papers were searched. When sperm samples from fertile and infertile males were analyzed, some of the mechanisms that determine the oxidative stress level were found to be impaired while others were unaltered. In conclusion, the literature as a whole provides contradictory findings and it is necessary to carry out more work to identify all the enzymatic and non-enzymatic systems involved in oxidative stress in the ejaculate, in order to develop new diagnostic systems and therapeutic strategies for combating detrimental free radical imbalance in the semen.

Antioxidants↗
PubMed2003

How does endometriosis affect infertility?

Prospective and retrospective clinical trials suggest a decreased oocyte and embryo quality in women with endometriosis. Based on these observations, the authors described an altered intrafollicular milieu in endometriosis, which explains the bad quality oocytes and the resulting embryos with lower capacity to implant. Whether these changes affect the oocytes or are the consequence of genomic alterations manifested by biochemical and chromosomal differences in healthy women is an unresolved issue. If the effects of endometriosis on follicular development are nongenomic in origin, modulation of the process of folliculogenesis may be sufficient to treat the disease and cure infertility associated with endometriosis. A genomic defect needs specific genetic therapy, which currently is not available.

Endometriosis↗
PubMed2003

Uterine and ovarian function in endometriosis.

Although numerous studies have attempted to elucidate the implication of endometriosis for infertility, this matter still remains unexplained. Diverse clinical approaches based on assisted reproduction techniques have produced conflicting results on this issue. Nevertheless, impairment of implantation and pregnancy rates seems to affect women who suffer from endometriosis. Whether this effect is due to alteration of the oocyte/embryo or the endometrium still remains to be elucidated, although actual data allude to deficiencies in both. Infertility due to poor quality embryos derived from impaired oocytes obtained from malfunctioning ovaries have been described by some groups. Endometrial defects have been argued by others. Adding more confusion to this topic, the influence of peritoneal fluid in women with endometriosis and/or mixed causes also needs to be considered. The aim of this review is to describe the current situation in the endometriosis-related infertility literature.

Ascitic Fluid↗
PubMed2002

Comparison of polymerase chain reaction-dependent methods for determining the presence of human immunodeficiency virus and hepatitis C virus in washed sperm.

OBJECTIVE(S): To evaluate the effectiveness of sperm washes by double capacitation in patients with human immunodeficiency virus (HIV) and hepatitis C virus (HCV) and to compare between polymerase chain reaction (PCR)-dependent methods to determine viral presence after the procedure. DESIGN: Controlled prospective clinical trial. SETTING: Private and university-affiliated setting. PATIENT(S): Thirty-four HIV-positive men attending our center for assisted reproduction with their partners. INTERVENTION(S): Forty-one semen samples from seropositive males were obtained and washed through density gradients and swim-up and analyzed for the presence of HIV and HCV with either nested or one-round PCR. Qualitative and quantitative detection of DNA and RNA of HIV and RNA of HCV. RESULT(S): Of all the semen samples that were analyzed after washing, seven out of 41 samples tested positive for one or both viruses by nested PCR. The confirmation of these results with the currently used commercial methods of quantitative PCR (one-round PCR) resulted in all negatives. CONCLUSION(S): Sperm wash with nested PCR is the appropriate method to use in the assisted reproduction techniques that are offered to serodiscordant couples. The detection limits exhibited by one-round PCR do not offer a sufficient guarantee that transmission of all viral particles will be avoided since both viruses can be present in amounts lower than the detection threshold of this technique.

Adult↗
PubMed2002

Gender selection: ethical, scientific, legal, and practical issues.

PURPOSE: This work offers an overview of the technology available and assisted reproductive technology (ART) practitioner attitudes to human reproductive sex preselection in either sperm with the use of flow cytometry (FC) or in embryos by means of preimplantational genetic diagnose (PGD) together with an analysis of Spanish legal environment. METHODS: A review of the legal, ethical and technical literature of the methods to select the sex in the offspring is performed. RESULTS: Sex selection in humans has different utilities to be employed depending on each country's law. Moreover, different ethical concerns are raised depending on the type of sex selection, in sperm or embryos. Both methodologies to pre-select the sex are trustworthy, with a high predictive power in the determination of the sex, but nevertheless, PGD is better than FC selection in this aspect. CONCLUSIONS: After a careful analysis of ethical, legal, and scientific features of gender selection, FC combined with PGD appears to be the most acceptable way to select the sex of the progeny whatever the circumstances are.

Female↗