We present our first experience with human embryo cryopreservation. In our hands the best results... more We present our first experience with human embryo cryopreservation. In our hands the best results were obtained with cryopreservation of pronuclear-staged embryos using propanediol as cryoprotectant. In this group 3 embryos out of 4 continued their development after thawing. The advantages of embryo cryopreservation in IVF programs are discussed.
graduated in Biological Science from Granada University in 2002. She has co-authored important re... more graduated in Biological Science from Granada University in 2002. She has co-authored important research articles in the field of assisted reproduction. She is responsible for Assisted Reproduction Laboratory at MAR&Gen Clinic in Granada. This matched case-controlled study shows that IVF failure associated with low serum progesterone levels after embryo transfer can be successfully treated by supporting the luteal phase with GnRH agonist.
Abstract The aim of this study was to evaluate the effect of two doses of d-chiro-inositol (DCI) ... more Abstract The aim of this study was to evaluate the effect of two doses of d-chiro-inositol (DCI) in combination with Myo-inositol (MYO) on the oocyte quality (OQ) of women with polycystic ovarian syndrome (PCOS) undergoing intracytoplasmic sperm injection (ICSI). Methods: This was a controlled, randomized, double-blind, parallel group study on 172 oocytes from 11 women. The study compared the effect of two MYO-DCI formulations given over 12 weeks on OQ. Five women received 550 mg of MYO + 300 mg of DCI daily (high DCI content group), while 6 women were given a daily dose of 550 mg of MYO with the only 27.6 mg of DCI (low DCI content group). Results: According to a multivariate analysis using linear mixed effect models, high doses of DCI have a positive influence on the quality of the cytoplasm of the oocyte (β = 1.631, χ2 = 7.347, d.f. = 1, p = .00672). Zona pellucida, plasma membrane, cytoplasm, and sperm reception have also been improved with any combination of MYO/DCI by decreasing testosterone or improving insulin sensitivity, regardless of age and body mass index. Conclusion: The combination of MYO with high doses of DCI improved oocyte cytoplasm quality in women with PCOS undergoing ICSI.
Round spermatids and ICSI logical cases, the usefulness of using round spermatids remains to be p... more Round spermatids and ICSI logical cases, the usefulness of using round spermatids remains to be proven on a large scale. Therefore, these data suggest spermatid injection should be considered as an option in case of unexpected absence of spermatozoa. In the case of patients with complete block of spermiogenesis, actually, round spermatids should not be proposed to the couple as a therapy. Couples entering this treatment should be advised about the safety and the extremely low efficacy of the procedure if round spermatids are used. Moreover, the risk of genetic transmission of Y chromosome deletions and of genomic imprinting anomalities should not be overlooked and care should be taken to avoid dramatic consequences of such pathologies. References
Human spermatozoa previously washed from seminal plasma and incubated in capacitating medium were... more Human spermatozoa previously washed from seminal plasma and incubated in capacitating medium were exposed to human follicular fluid (FF) for different time periods and their movement characteristics were examined by computer-assisted digital image analysis. No differences from control samples were observed after short exposures (15 minutes and 2 hours). When the exposure was prolonged to 6 hours, spermatozoa incubated in the presence of FF displayed significant differences in some movement characteristics as compared with samples incubated without FF supplement. When only rapidly moving sperm subpopulations were considered, these effects of FF were similar to those produced by a short incubation with the solubilized cumulus oophorus matrix as described previously. After 24 hours of incubation, there was a remarkable beneficial effect of FF on motility preservation. Possible physiological and clinical implications are discussed.
Most studies published so far suggest that the administration of growth hormone (GH) during ovari... more Most studies published so far suggest that the administration of growth hormone (GH) during ovarian stimulation improves in vitro fertilization (IVF) outcomes, especially in older women, but also in some younger ones who suffered repeated IVF failures during their previous IVF attempts. However, clearly defined criteria with which to distinguish between those women who are likely to benefit from GH treatment and those who are not are still lacking. This minireview resumes the latest advances as to the indication of GH for female infertility treatment. A small case series, showing one way how to address this question, is also included.
The changes in acrosin immunoreactivity in human spermatozoa undergoing spontaneous or chemically... more The changes in acrosin immunoreactivity in human spermatozoa undergoing spontaneous or chemically induced acrosome reactions were studied by electron microscopic immunocytochemistry with an acrosin-specific monoclonal antibody. Migration of limited amounts of acrosin to the sperm surface was the earliest event characterizing the beginning of the acrosome reaction. The acrosome of such spermatozoa remained morphologically intact, swelled, or showed intraacrosomal vesiculation without any disruption of the plasma and acrosomal membrane integrity. Massive release of acrosin coincided with the fusion of the plasma and outer acrosomal membranes. However, even fully acrosomereacted spermatozoa always retained some acrosin on the exposed inner acrosomal membrane and in the equatorial segment of the acrosome. This residual acrosin was also detected on spermatozoa within the zona pellucida of human oocytes inseminated in vitro, while the previously released bulk of acrosin remained attached to the surface of the zona pellucida at the site of sperm entry. These findings are compatible with multiple functions of acrosin in human sperm-egg interaction, including sperm-zona pellucida binding, dispersal of acrosomal contents, and facilitation of zona pellucida penetration. Fertil Steril 50:133, 1988 Acrosin is the most studied protease of mammalian spermatozoa. During human spermatogenesis it is first expressed in round spermatids at the beginning of the cap phase of acrosomal development, 1 and, up to the time of fertilization, it is believed to be sequestered within the sperm acrosome where it exists in the zymogen form-proacrosin. 2 While there is a general consent as to the notion that the conversion of proacrosin into acrosin
Obstetrics & Gynecology International Journal, Jan 16, 2020
Menopause, natural or premature, was long been considered to be incompatible with conception with... more Menopause, natural or premature, was long been considered to be incompatible with conception with the own eggs of the women affected. Here we describe a case of a 32-year-old woman with premature menopause, characterized by amenorrea, the absence of antral follicles in the ovaries, high basal follicle-stimulating hormone (FSH) and luteinizing hormone (LH) levels and extremely low antimullerian hormone (AMH) levels. After preliminary treatment with contraceptive pill for two months, and an injection of recombinant human chorionic gonadotropin (HCG) to boost intrinsic androgen production, ovarian stimulation, using a customized protocol with a patient-tailored combination of recombinant FSH and LH, in addition to daily injections of growth hormone, resulted in the recovery of 3 metaphase II eggs after double triggering with recombinant HCG. After transfer of two embros, obtained by intracytoplasmic sperm injection, an ongoing twin clinical pregnancy was achieved. These data show that oocyte recovery, fertilization, embryo development and pregnancy can be achieved, with the use of a customized ovarian stimulation protocol, even in young women with clear signs of advanced premature menopause.
between fertile and subfertile. Semen analysis was performed according to the WHO guidelines, exc... more between fertile and subfertile. Semen analysis was performed according to the WHO guidelines, except for sperm morphology, where strict criteria were used. The fertile population was defined by a pregnancy occurring within 12 months, whereas the subfertile controls were obtained from 137 consecutive couples attending our fertility clinic for the first time. Only couples with no tubal or ovulatory factor were included in the study. Biochemical factors such as ILl -, dimeric inhibin B, transferrin, soluble transferrin receptor, xanthine oxidase, hepatocyte growth factor, Clara cell protein (CC 16), interleukin 7, and the LIF receptor were measured through sensitive ELISA methods. The a-glucosidase activity was determined using the method described by Chapdelaine et al.
International Journal of Gynecology & Obstetrics, 2021
To examine whether adding a second HCG trigger, 12.5 h after the first (36.5 h before ovarian pun... more To examine whether adding a second HCG trigger, 12.5 h after the first (36.5 h before ovarian puncture), can facilitate recovery of oocytes in women with a paucifollicular response to ovarian stimulation.
The aim of this review is to provide an overview of the literature about the perception and manag... more The aim of this review is to provide an overview of the literature about the perception and management of anxiety and pain in women undergoing an office hysteroscopic procedure. Methods We performed a systematic literature search in Embase, PubMed/MEDLINE, Cochrane Library and Web of Science for original studies written in English (registered in PROSPERO 2019-CRD42019132341), using the terms 'hysteroscopy' AND 'pain' AND 'anxiety' published up to January 2019. Only original articles (randomized, observational and retrospective studies) about management of anxiety and pain related to the hysteroscopic procedure were considered eligible. Results Our literature search produced 84 records. After exclusions, 11 studies including 2222 patients showed the following results: (a) pain experienced during hysteroscopy is negatively affected by preprocedural anxiety; (b) pharmacological interventions seem to be help in reducing pain during hysteroscopy; (c) waiting time before the procedure is a significant factor affecting patients' anxiety; (d) music during the procedure may be helpful in reducing anxiety. Conclusions The utilization of office hysteroscopy is hampered by varying levels of anxiety and pain perceived by women who are candidates for the procedure. For these reasons, it is essential to identify effective pharmacological and non-pharmacological strategies to alleviate these factors. We recommend further studies especially focusing on non-pharmacological interventions to facilitate the dissemination of good clinical practices among hysteroscopists.
Evidence has accumulated suggesting multiple roles of acrosin in fertilization, including its par... more Evidence has accumulated suggesting multiple roles of acrosin in fertilization, including its participation in early steps of gamete recognition and binding. However, the implication of acrosin in many of these processes is not compatible with its presumptive sequestration within the sperm acrosome until a late phase of the acrosome reaction. In an earlier study (J. Tesarik, J. Drahorad, J. Peknicova, 1988, Fertil. Steril. 50, 133-141), we reported the binding of an anti-acrosin monoclonal antibody (M0-AKR.1) to the plasma membrane overlying the acrosome of human spermatozoa starting the acrosome reaction. In this study, we characterized further this antibody with regard to its reactivity with different forms of acrosin and found that it recognizes specifically an active form of this enzyme and does not react with its proenzyme form. MO-AKR.l was thus used as a probe for in situ analysis of acrosin activation during the acrosome reaction. When suspensions of living spermatozoa were ...
Current Opinion in Obstetrics & Gynecology, 2020
Purpose of review The aim of this systematic review is to summarize the current evidence regardin... more Purpose of review The aim of this systematic review is to summarize the current evidence regarding the effectiveness of hysterectomy and hysteroscopic endometrial resection in improving quality of life (QoL), sexual function and psychological wellbeing of women abnormal uterine bleeding. Recent findings We performed a systematic literature search in PubMed/MEDLINE and Embase for original studies written in English (registered in PROSPERO 2019 CRD42019133632), using the terms ‘endometrial ablation’, ‘endometrial destruction’, ‘endometrial resection’, ‘hysterectomy’, ‘menorrhagia’, ‘dysfunctional uterine bleeding’, ‘quality of life’, ‘sexuality’ published up to April 2019. Our literature search produced 159 records. After exclusions, nine studies were included showing the following results: both types of treatment significantly improve QoL and psychological wellbeing; hysterectomy is associated with higher rates of satisfaction; hysterectomy is not associated with a significant deteri...
We present our first experience with human embryo cryopreservation. In our hands the best results... more We present our first experience with human embryo cryopreservation. In our hands the best results were obtained with cryopreservation of pronuclear-staged embryos using propanediol as cryoprotectant. In this group 3 embryos out of 4 continued their development after thawing. The advantages of embryo cryopreservation in IVF programs are discussed.
graduated in Biological Science from Granada University in 2002. She has co-authored important re... more graduated in Biological Science from Granada University in 2002. She has co-authored important research articles in the field of assisted reproduction. She is responsible for Assisted Reproduction Laboratory at MAR&Gen Clinic in Granada. This matched case-controlled study shows that IVF failure associated with low serum progesterone levels after embryo transfer can be successfully treated by supporting the luteal phase with GnRH agonist.
Abstract The aim of this study was to evaluate the effect of two doses of d-chiro-inositol (DCI) ... more Abstract The aim of this study was to evaluate the effect of two doses of d-chiro-inositol (DCI) in combination with Myo-inositol (MYO) on the oocyte quality (OQ) of women with polycystic ovarian syndrome (PCOS) undergoing intracytoplasmic sperm injection (ICSI). Methods: This was a controlled, randomized, double-blind, parallel group study on 172 oocytes from 11 women. The study compared the effect of two MYO-DCI formulations given over 12 weeks on OQ. Five women received 550 mg of MYO + 300 mg of DCI daily (high DCI content group), while 6 women were given a daily dose of 550 mg of MYO with the only 27.6 mg of DCI (low DCI content group). Results: According to a multivariate analysis using linear mixed effect models, high doses of DCI have a positive influence on the quality of the cytoplasm of the oocyte (β = 1.631, χ2 = 7.347, d.f. = 1, p = .00672). Zona pellucida, plasma membrane, cytoplasm, and sperm reception have also been improved with any combination of MYO/DCI by decreasing testosterone or improving insulin sensitivity, regardless of age and body mass index. Conclusion: The combination of MYO with high doses of DCI improved oocyte cytoplasm quality in women with PCOS undergoing ICSI.
Round spermatids and ICSI logical cases, the usefulness of using round spermatids remains to be p... more Round spermatids and ICSI logical cases, the usefulness of using round spermatids remains to be proven on a large scale. Therefore, these data suggest spermatid injection should be considered as an option in case of unexpected absence of spermatozoa. In the case of patients with complete block of spermiogenesis, actually, round spermatids should not be proposed to the couple as a therapy. Couples entering this treatment should be advised about the safety and the extremely low efficacy of the procedure if round spermatids are used. Moreover, the risk of genetic transmission of Y chromosome deletions and of genomic imprinting anomalities should not be overlooked and care should be taken to avoid dramatic consequences of such pathologies. References
Human spermatozoa previously washed from seminal plasma and incubated in capacitating medium were... more Human spermatozoa previously washed from seminal plasma and incubated in capacitating medium were exposed to human follicular fluid (FF) for different time periods and their movement characteristics were examined by computer-assisted digital image analysis. No differences from control samples were observed after short exposures (15 minutes and 2 hours). When the exposure was prolonged to 6 hours, spermatozoa incubated in the presence of FF displayed significant differences in some movement characteristics as compared with samples incubated without FF supplement. When only rapidly moving sperm subpopulations were considered, these effects of FF were similar to those produced by a short incubation with the solubilized cumulus oophorus matrix as described previously. After 24 hours of incubation, there was a remarkable beneficial effect of FF on motility preservation. Possible physiological and clinical implications are discussed.
Most studies published so far suggest that the administration of growth hormone (GH) during ovari... more Most studies published so far suggest that the administration of growth hormone (GH) during ovarian stimulation improves in vitro fertilization (IVF) outcomes, especially in older women, but also in some younger ones who suffered repeated IVF failures during their previous IVF attempts. However, clearly defined criteria with which to distinguish between those women who are likely to benefit from GH treatment and those who are not are still lacking. This minireview resumes the latest advances as to the indication of GH for female infertility treatment. A small case series, showing one way how to address this question, is also included.
The changes in acrosin immunoreactivity in human spermatozoa undergoing spontaneous or chemically... more The changes in acrosin immunoreactivity in human spermatozoa undergoing spontaneous or chemically induced acrosome reactions were studied by electron microscopic immunocytochemistry with an acrosin-specific monoclonal antibody. Migration of limited amounts of acrosin to the sperm surface was the earliest event characterizing the beginning of the acrosome reaction. The acrosome of such spermatozoa remained morphologically intact, swelled, or showed intraacrosomal vesiculation without any disruption of the plasma and acrosomal membrane integrity. Massive release of acrosin coincided with the fusion of the plasma and outer acrosomal membranes. However, even fully acrosomereacted spermatozoa always retained some acrosin on the exposed inner acrosomal membrane and in the equatorial segment of the acrosome. This residual acrosin was also detected on spermatozoa within the zona pellucida of human oocytes inseminated in vitro, while the previously released bulk of acrosin remained attached to the surface of the zona pellucida at the site of sperm entry. These findings are compatible with multiple functions of acrosin in human sperm-egg interaction, including sperm-zona pellucida binding, dispersal of acrosomal contents, and facilitation of zona pellucida penetration. Fertil Steril 50:133, 1988 Acrosin is the most studied protease of mammalian spermatozoa. During human spermatogenesis it is first expressed in round spermatids at the beginning of the cap phase of acrosomal development, 1 and, up to the time of fertilization, it is believed to be sequestered within the sperm acrosome where it exists in the zymogen form-proacrosin. 2 While there is a general consent as to the notion that the conversion of proacrosin into acrosin
Obstetrics & Gynecology International Journal, Jan 16, 2020
Menopause, natural or premature, was long been considered to be incompatible with conception with... more Menopause, natural or premature, was long been considered to be incompatible with conception with the own eggs of the women affected. Here we describe a case of a 32-year-old woman with premature menopause, characterized by amenorrea, the absence of antral follicles in the ovaries, high basal follicle-stimulating hormone (FSH) and luteinizing hormone (LH) levels and extremely low antimullerian hormone (AMH) levels. After preliminary treatment with contraceptive pill for two months, and an injection of recombinant human chorionic gonadotropin (HCG) to boost intrinsic androgen production, ovarian stimulation, using a customized protocol with a patient-tailored combination of recombinant FSH and LH, in addition to daily injections of growth hormone, resulted in the recovery of 3 metaphase II eggs after double triggering with recombinant HCG. After transfer of two embros, obtained by intracytoplasmic sperm injection, an ongoing twin clinical pregnancy was achieved. These data show that oocyte recovery, fertilization, embryo development and pregnancy can be achieved, with the use of a customized ovarian stimulation protocol, even in young women with clear signs of advanced premature menopause.
between fertile and subfertile. Semen analysis was performed according to the WHO guidelines, exc... more between fertile and subfertile. Semen analysis was performed according to the WHO guidelines, except for sperm morphology, where strict criteria were used. The fertile population was defined by a pregnancy occurring within 12 months, whereas the subfertile controls were obtained from 137 consecutive couples attending our fertility clinic for the first time. Only couples with no tubal or ovulatory factor were included in the study. Biochemical factors such as ILl -, dimeric inhibin B, transferrin, soluble transferrin receptor, xanthine oxidase, hepatocyte growth factor, Clara cell protein (CC 16), interleukin 7, and the LIF receptor were measured through sensitive ELISA methods. The a-glucosidase activity was determined using the method described by Chapdelaine et al.
International Journal of Gynecology & Obstetrics, 2021
To examine whether adding a second HCG trigger, 12.5 h after the first (36.5 h before ovarian pun... more To examine whether adding a second HCG trigger, 12.5 h after the first (36.5 h before ovarian puncture), can facilitate recovery of oocytes in women with a paucifollicular response to ovarian stimulation.
The aim of this review is to provide an overview of the literature about the perception and manag... more The aim of this review is to provide an overview of the literature about the perception and management of anxiety and pain in women undergoing an office hysteroscopic procedure. Methods We performed a systematic literature search in Embase, PubMed/MEDLINE, Cochrane Library and Web of Science for original studies written in English (registered in PROSPERO 2019-CRD42019132341), using the terms 'hysteroscopy' AND 'pain' AND 'anxiety' published up to January 2019. Only original articles (randomized, observational and retrospective studies) about management of anxiety and pain related to the hysteroscopic procedure were considered eligible. Results Our literature search produced 84 records. After exclusions, 11 studies including 2222 patients showed the following results: (a) pain experienced during hysteroscopy is negatively affected by preprocedural anxiety; (b) pharmacological interventions seem to be help in reducing pain during hysteroscopy; (c) waiting time before the procedure is a significant factor affecting patients' anxiety; (d) music during the procedure may be helpful in reducing anxiety. Conclusions The utilization of office hysteroscopy is hampered by varying levels of anxiety and pain perceived by women who are candidates for the procedure. For these reasons, it is essential to identify effective pharmacological and non-pharmacological strategies to alleviate these factors. We recommend further studies especially focusing on non-pharmacological interventions to facilitate the dissemination of good clinical practices among hysteroscopists.
Evidence has accumulated suggesting multiple roles of acrosin in fertilization, including its par... more Evidence has accumulated suggesting multiple roles of acrosin in fertilization, including its participation in early steps of gamete recognition and binding. However, the implication of acrosin in many of these processes is not compatible with its presumptive sequestration within the sperm acrosome until a late phase of the acrosome reaction. In an earlier study (J. Tesarik, J. Drahorad, J. Peknicova, 1988, Fertil. Steril. 50, 133-141), we reported the binding of an anti-acrosin monoclonal antibody (M0-AKR.1) to the plasma membrane overlying the acrosome of human spermatozoa starting the acrosome reaction. In this study, we characterized further this antibody with regard to its reactivity with different forms of acrosin and found that it recognizes specifically an active form of this enzyme and does not react with its proenzyme form. MO-AKR.l was thus used as a probe for in situ analysis of acrosin activation during the acrosome reaction. When suspensions of living spermatozoa were ...
Current Opinion in Obstetrics & Gynecology, 2020
Purpose of review The aim of this systematic review is to summarize the current evidence regardin... more Purpose of review The aim of this systematic review is to summarize the current evidence regarding the effectiveness of hysterectomy and hysteroscopic endometrial resection in improving quality of life (QoL), sexual function and psychological wellbeing of women abnormal uterine bleeding. Recent findings We performed a systematic literature search in PubMed/MEDLINE and Embase for original studies written in English (registered in PROSPERO 2019 CRD42019133632), using the terms ‘endometrial ablation’, ‘endometrial destruction’, ‘endometrial resection’, ‘hysterectomy’, ‘menorrhagia’, ‘dysfunctional uterine bleeding’, ‘quality of life’, ‘sexuality’ published up to April 2019. Our literature search produced 159 records. After exclusions, nine studies were included showing the following results: both types of treatment significantly improve QoL and psychological wellbeing; hysterectomy is associated with higher rates of satisfaction; hysterectomy is not associated with a significant deteri...
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Papers by Jan Tesarik