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1.
Genesis ; 58(3-4): e23350, 2020 03.
Article in English | MEDLINE | ID: mdl-31815354

ABSTRACT

A previous animal study by our group found that sleep deprivation during preimplantation was associated with decreased pregnancy maintenance. Given its impact on human society, we aimed in the current study to assess whether sleep deprivation affects blastocyst gene expression and/or the implantation process. For this, pregnant mice (gestational day 0 [GD 0]) were assigned into paradoxical sleep deprivation (SD, 72 hr; multiple platform method) and, a control (CT) group. Animals were euthanized on GD 3.5 and blood, uterus (embryos) and fallopian tube were collected. Then, 89% of CT presented blastocysts in the uterus versus 25% from SD group. Compared to CT, SD presented lighter relative uterus weight, increased plasma concentrations of corticosterone and testosterone, decreased concentrations of progesterone and luteinizing hormone, but no statistical differences in plasma concentrations of 17ß-estradiol and follicle stimulating hormone. There were no differences in uterus and blastocyst gene expression related to embryo implantation and development, and no alteration in blastocysts global DNA methylation. Considering this, the decreased pregnancy maintenance after sleep deprivation seems not to be associated with implantation losses or developmental problems related to the blastocysts. It is likely that complications in morula development and/or its movement through the fallopian tubes affect the pregnancy rate, since only 25% of SD females presented a blastocyst on the GD 3.5. In fact, three out of four females without blastocysts in the uterus presented morula in the fallopian tubes due to a phase delay. Additionally, we suggest that the observed hormonal changes may play a role in this outcome.


Subject(s)
Embryo Implantation , Morula/metabolism , Reproduction , Sleep Deprivation , Uterus/physiology , Animals , Biomarkers , Blastocyst/metabolism , Body Weight , DNA Methylation , Epigenesis, Genetic , Fallopian Tubes/metabolism , Female , Fluorescent Antibody Technique , Gene Expression Regulation , Hormones/metabolism , Mice , Time Factors
2.
JBRA Assist Reprod ; 22(1): 71-74, 2018 Mar 01.
Article in English | MEDLINE | ID: mdl-29345446

ABSTRACT

Sickle cell anemia is an inherited systemic hemoglobinopathy that affects hemoglobin production in red blood cells, leading to early morbidity and mortality. It is caused by a homozygous nucleotide substitution (c.20A>T) in the ß-globin gene (HBB) that changes a glutamic acid to a valine in the protein. We present a case report of a fertile couple, both carriers of the sickle cell anemia mutation, with one affected daughter. Six cycles of assisted reproductive techniques were performed, resulting in 53 embryos in cleavage stage. Each embryo was biopsied and analyzed for pre-implantation genetic diagnosis (PGD) by fluorescent polymerase chain reaction, using polymorphic markers of the region of interest followed by capillary electrophoresis in an automated genetic analyzer. HLA Compatible and normal embryos for the mutation represented 3 (5.66%); while the carriers and compatible 6 (11.32%); therefore, embryos matching those of the affected daughter represented 9 (16.98%). A selected embryo in blastocyst stage was transferred, resulting in a healthy male newborn, who had the umbilical cord blood cells collected and stored. The affected daughter was immunosuppressed and received transplanted cells from the umbilical cord blood of her brother; the treatment was successful. Embryo selection using PGD technologies represent the most effective treatment plan for parents who want to have a healthy child, and it could cure another child already affected by inherited hemoglobinopathy.


Subject(s)
Anemia, Sickle Cell/therapy , Cord Blood Stem Cell Transplantation , Histocompatibility Testing/methods , Preimplantation Diagnosis/methods , Adult , Anemia, Sickle Cell/blood , Anemia, Sickle Cell/genetics , Blastocyst/metabolism , Blastocyst/pathology , Child , Embryo Transfer , Female , Genetic Linkage , Genetic Testing/methods , Humans , Infant, Newborn , Male , Pedigree , Pregnancy , Reproductive Techniques, Assisted , Sibling Relations
3.
Fertil Steril ; 106(2): 273-283.e6, 2016 Aug.
Article in English | MEDLINE | ID: mdl-27105718

ABSTRACT

OBJECTIVE: To study the effects of n-6 and n-3 polyunsaturated acid-rich soybean phosphatidylcholine (soy-PC) on sperm cryotolerance with regard to sperm membrane lipid profile, membrane surface integrity, and routine semen parameters. DESIGN: Experimental study. SETTING: University-affiliated tertiary hospital. PATIENT(S): A total of 20 normospermic fertile men. INTERVENTION(S): Semen samples examined for differences in semen parameters, sperm membrane lipid profile, and plasma membrane surface both before and after cryopreservation using basic freezing medium with N-tris(hydroxymethyl)-methyl-2-aminoethane sulfonic acid (TES) and tris-(hydroxymethyl)-aminomethane (TRIS) supplemented with purified soy-PC (TEST-PC) or egg yolk (TEST-Y), both alone or in association (TEST-Y-PC). MAIN OUTCOME MEASURE(S): Conventional semen parameters and membrane lipid profile by matrix-assisted laser/desorption ionization mass spectrometry (MALDI-MS). RESULT(S): Postthaw sperm cell motility, vitality, and morphology parameters were similar for soy-PC (TEST-PC) and egg yolk (TEST-Y) cryoprotectants. However, sperm exposed to TEST-Y-PC presented better kinetic parameters, which were similar to the original quality of the fresh semen. Human sperm MALDI-MS lipid profiles revealed that the relative abundance of glycerophospholipids of m/z 760.44 [PC (34:1)+H]+, 781.55 [SM (20:0) +Na]+, 784.55 [PC (36:3) +H]+, 806.64 [PC (38:6) +H]+, 807.64 [SM (22:1) +Na]+, and 809.64 [SM (22:0) +Na]+ increased in soy-PC samples (TEST-PC). Nonetheless, only one lipid (m/z 781.55, [SM (20:0) +Na]+) statistically significantly changed when sperm was cryopreserved in TEST-Y-PC. CONCLUSION(S): Sphingomyelin was defined as a prospective biomarker of soy-PC treatment, and it could be related to the positive cryoprotective effects of soy-PC in human sperm, opening new perspectives to design of a more efficient synthetic cryoprotectant medium containing purified egg yolk biomolecules combined with soy-PC.


Subject(s)
Cell Membrane/drug effects , Cold Temperature/adverse effects , Cryopreservation/methods , Cryoprotective Agents/pharmacology , Fatty Acids, Omega-3/pharmacology , Fatty Acids, Omega-6/pharmacology , Glycine max/chemistry , Membrane Lipids/metabolism , Phosphatidylcholines/pharmacology , Spermatozoa/drug effects , Cell Membrane/metabolism , Cell Membrane/ultrastructure , Cell Survival/drug effects , Cryoprotective Agents/isolation & purification , Egg Yolk/chemistry , Fatty Acids, Omega-3/isolation & purification , Fatty Acids, Omega-6/isolation & purification , Humans , Kinetics , Male , Micelles , Microscopy, Electron, Scanning , Phosphatidylcholines/isolation & purification , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Sperm Motility/drug effects , Spermatozoa/metabolism , Spermatozoa/ultrastructure , Sphingomyelins/isolation & purification , Sphingomyelins/pharmacology
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