Concentration of glutathione and expression of glutathione peroxidases 1 and 4 in fresh sperm provide a forecast of the outcome of cryopreservation of human spermatozoa

Oxidative stress imbalance potentially leads to damage of the structure of the cell and macromolecules such as plasma membrane components, proteins, and DNA. The plasma membrane of the sperm cell, which has high levels of polyunsaturated fatty acids, renders it particularly sensitive to free radical–mediated attacks. The freezing and subsequent thawing of sperm is a physically stressful process carried out during routine procedures in assisted reproduction techniques, which results in a highly variable and unpredictable reduction in the number of motile sperm cells. Subsequently, oxidative status can positively or negatively affect the motility, viability, and fertilizing capacity of thawed sperm. These effects are counteracted by various oxidative defense enzymes and anti-oxidants such as glutathione peroxidase isoforms GPx1 and GPx4, glutathione reductase (GR), and cellular glutathione (reduced) (GSH). In this way, oxidative status could represent a predictive marker of sperm quality following the freeze-thaw process. This study was based on 56 human sperm samples. We observed direct positive and negative relationships between the postthaw motile sperm recovery rate and GPx1 and GPx4 expression and activity, on the one hand, and GSH concentrations, on the other. No correlation was found between this recovery rate and GR or basic semen parameters. Predictive values clearly demonstrate that, among the molecules analyzed, the most accurate diagnoses result when analyses are conducted for GPx1 and GPx1 messenger RNA expression, GPx1 and GPx4 enzymatic activity, and GSH concentration. In conclusion, a reserve of glutathione, together with GPx expression, is necessary to eliminate free radicals using GSH or a like structural protein and seems to be essential for a good postthaw recovery. These molecules can be employed as indicators of postthaw sperm quality.Meseguer Anastasio, Manuel Francisco, [email protected] ; Simon Valles, Carlos Antonio, [email protected] ; Remohi Gimenez, Jose Alejandro, [email protected]

Comparison of different sources of platelet-rich plasma as treatment option for infertility-causing endometrial pathologies

Objective. To study the effect of human plasma from different sources such as umbilical cord blood and adult blood platelet-rich plasma (PRP) on the regeneration of endometrial damage. Design. Composition analysis, in vitro approaches and a pre-clinical murine model using plasma to promote endometrial regeneration. Setting. Hospital and university laboratories. Patients/Animals. Adult plasma from 4 Atrophy patients and one fertile woman, commercial umbilical cord plasma and uterine-damaged NOD/SCID mice model were used. Intervention(s). Endometrial stromal cells from primary culture and an endometrial stem cell line were cultured in vitro and uterine-damaged NOD/SCID mice were treated with plasma samples from several origins. Main Outcome Measure(s). All plasma samples contain molecules with a high potential for regeneration (SCF, PDGFBB, THBS1, VWF). Furthermore, the highest increase in in vitro proliferation and migration rate was found when endometrial stromal cells were treated with umbilical cord plasma, adult PRP also revealed a significant increment. In the mouse model, a higher expression of Ki67 and Hoxa10 in the endometrium was detected after applying adult PRP and the proteomic analysis revealed a specific protein expression profile depending on the treatment. The damaged uterine tissue showed more pro22 regenerative markers after applying umbilical cord plasma (Stat5a, Uba3, Thy1) in comparison to the other treatments (non-activated umbilical cord plasma, activated adult PRP and not treatment). Conclusion. Human PRP possesses regeneration properties usable for endometrial pathologies. Besides that, these regenerative effects seem to be more apparent when the source of obtaining is umbilical cord blood

High incidence of chromosomal abnormalities in large-headed and multiple-tailed spermatozoa

Rodrigo Vivo, Lorena, [email protected] ; Prados Dodd, Nicolas, [email protected] ; Gil Salom, Manuel Luis, [email protected] ; Remohi Gimenez, Jose Alejandro, [email protected]

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Number needed to freeze: cumulative live birth rate after fertility preservation in women with endometriosis

Research question: How does the number of oocytes used affect the cumulative live birth rate in endometriosis patients who had their oocytes vitrified for fertility preservation (FP)? Design: Retrospective observational study including data from 485 women with endometriosis who underwent FP from January 2007 to July 2018. Survival curves and Kaplan-Meier plots were used to analyse the cumulative live birth rate (CLBR) according to the number of vitrified oocytes used. Data were stratified according to age, stage of the disease and ovarian surgery prior to FP (operated vs. non-operated). Endometriosis curves were compared to plots developed using elective fertility preservation (EFP) patients as control group. Log-rank, Breslow and Tarone-Ware tests were used to compare the survival curves. Results: The CLBR increased as the number of oocytes used per patient rose, reaching 89.5% (95% CI=80.0-99.1) using 22 oocytes. Higher outcomes were observed in young women (≤35 y. vs. >35 y). In the younger group, the CLBR was 95.4% (95% CI=87.2-103.6) using ~20 oocytes vs. 79.6% (95% CI=58.1-101.1) in older women (P<0.05). No statistical differences were observed in overall calculations and according to age when the CLBR was compared between operated and non-operated women (NS). Comparable outcomes were also observed in stages I-II vs. III-IV (NS). The mean age was higher in EFP patients (37.2 ± 4.9 vs. 35.7 ± 3.7; P<0.05). The outcome was better in the endometriosis group as compared to EFP (P<0.05): a CLBR of 89.5% (80.0-99.1) vs. 59.9% (51.4-68.6) when 22 oocytes were used (P<0.05). However, the difference was milder when fewer oocytes were used in both groups. When comparisons were made between age-matching groups, no statistical differences were observed (NS). Conclusion: The probability of live birth increases as the number of oocytes used rises in patients with endometriosis, but better outcomes were observed among young women. Neither the stage of the disease nor prior surgical excision of ovarian endometrioma were related to success. No statistical differences in age matching groups were observed when comparing to EFP patients. The information provided herein may be of interest to both patients and treating physicians for counselling purposes

Stem cell paracrine actions in tissue regeneration and its potential therapeutic effect in human endometrium: a retrospective study.

Objective: Determining genetic and paracrine mechanisms behind endometrial regeneration in Asherman's Syndrome and Endometrial Atrophy (AS/EA) patients after autologous CD133+ bone marrow-derived stem cells (CD133+BMDSCs) transplantation. Design: Retrospective study using human endometrial biopsies and mouse models. Setting: Fundación-IVI, IIS-La Fe, Valencia, Spain. Samples: Endometrial biopsies collected before and after CD133+BMDSCs therapy, from 8 women with AS/EA (NCT02144987). And uterus from 5 mice, with only left horns receiving CD133+BMDSCs therapy. Methods: In human samples, hematoxylin and eosin (H&E) staining, RNA arrays, PCR validation and neutrophil elastase (NE) immunohistochemistry (IHQ). In mouse samples, PCR validation and protein immunoarrays

The Lin28/Let-7 System in Early Human Embryonic Tissue and Ectopic Pregnancy

Our objective was to determine the expression of the elements of the Lin28/Let-7 system, and related microRNAs (miRNAs), in early stages of human placentation and ectopic pregnancy, as a means to assess the potential role of this molecular hub in the pathogenesis of ectopic gestation. Seventeen patients suffering from tubal ectopic pregnancy (cases) and forty-three women with normal on-going gestation that desired voluntary termination of pregnancy (VTOP; controls) were recruited for the study. Embryonic tissues were subjected to RNA extraction and quantitative PCR analyses for LIN28B, Let-7a, miR-132, miR-145 and mir-323-3p were performed. Our results demonstrate that the expression of LIN28B mRNA was barely detectable in embryonic tissue from early stages of gestation and sharply increased thereafter to plateau between gestational weeks 7–9. In contrast, expression levels of Let-7, mir-132 and mir-145 were high in embryonic tissue from early gestations (≤6-weeks) and abruptly declined thereafter, especially for Let-7. Opposite trends were detected for mir-323-3p. Embryonic expression of LIN28B mRNA was higher in early stages (≤6-weeks) of ectopic pregnancy than in normal gestation. In contrast, Let-7a expression was significantly lower in early ectopic pregnancies, while miR-132 and miR-145 levels were not altered. Expression of mir-323-3p was also suppressed in ectopic embryonic tissue. We are the first to document reciprocal changes in the expression profiles of the gene encoding the RNA-binding protein, LIN28B, and the related miRNAs, Let-7a, mir-132 and mir-145, in early stages of human placentation. This finding suggests the potential involvement of LIN28B/Let-7 (de)regulated pathways in the pathophysiology of ectopic pregnancy in humans

Manejo quirúrgico de la endometriosis vésico-ureteral y sus complicaciones

La endometriosis ureteral es una infrecuente localización de endometriosis profunda, que puede condicionar una grave disminución de la función renal de forma silenciosa. Se presenta el caso de una paciente con fibrosis peritoneal secundaria a endometriosis profunda, cuya inespecífica sintomatologia conllevó un retraso diagnóstico, permitiendo el desarrollo de hidronefrosis. Es necesario descartar la presencia de endometriosis profunda en mujeres en edad fértil con hidronefrosis de etiología desconocida

False positive rate of an arrayCGH platform for single-cell preimplantation genetic screening and subsequent clinical application on day-3

In this work, false positive rate of an arrayCGH platform for its use in day-3 single-blastomere analysis was calculated. For this purpose, 38 embryos diagnosed as abnormal on day-3 by FISH were re-biopsied on day-4. Single-cell day-4 arrayCGH diagnosis was then performed. A successful amplification was obtained in 97.4 % (37/38) of the day-4 cells analysed by arrayCGH. Day-3 FISH and day-4 arrayCGH diagnosis were concordant in 35/37 cases. The two discordant embryos were spread and all the cells from each embryo were re-analysed by FISH on day 5. The same error rate (2.7 %) for day-3 FISH and day-4 arrayCGH was obtained when comparing day-5 FISH re-analysis. After this pre-clinical phase, the platform was used for day-3 arrayCGH clinical application in 320 patients (1,760 embryos). Day-3 amplification rate was 98.6 %. An optimal reproductive outcome was obtained when applying arrayCGH to a clinical program: clinical pregnancy rate per cycle of 38.4 % and 60.3 % per transference were obtained, with an implantation rate of 53.5 %. Overall miscarriage rate was 10.6 %. Additionally, day-5 FISH re-analysis was performed in 42 of the embryos from the clinical phase, obtaining a concordance rate of 97.6 % with day-3 arrayCGH.Electronic supplementary materialThe online version of this article (doi:10.1007/s10815-012-9918-4) contains supplementary material, which is available to authorized users

Removing of Direct Red 83:1 using α- and HP-α-CDs polymerized with epichlorohydrin: Kinetic and equilibrium studies

The adsorption properties of two cyclodextrin-epichlorohydrin polymers to remove Direct Red 83:1 textile dye from wastewater solutions was evaluated. Adsorption studies were performed in a batch reactor, and the efficiency of the polymers under different parametric values such as contact time, adsorbent dosage, initial dye concentration, pH of initial solution and temperature was followed by spectrophotometry. Both Langmuir and Freundlich equations were used to fit the equilibrium isotherms and the kinetic data were analyzed by the pseudo-first-order and pseudo-second-order models. Adsorption process follows a pseudo-second-order kinetic and was influenced by intraparticle diffusion model, dye concentration and contact time, reaching equilibrium within the first 30 min. Experimental data were better fitted to Freundlich isotherm model than to Langmuir and Temkin isotherms. Since about 93% of dye removal was achieved under optimized conditions, the α-cyclodextrin-epichlorohydrin polymer seems to be a useful device for removing Direct Red 83:1 from aqueous solutions and industrial effluents.Ciencias Ambientale