Associations of fertility parameters with fatty acids and DNA methylation in Mexican women undergoing in vitro fertilization.

BACKGROUND: Fatty acids (FA) likely affect human fertility at multiple levels, as deviations from physiological FA profiles are obesogenic, and FA can modify DNA methylation (DNAm). Yet, the interplay of follicular fluid (FF) and serum FA with BMI and percentage body fat (PBF) in human fertility is not completely understood. Also, associations of DNAm with fertility are largely unexplored. METHODS: Reproductive parameters ranging from retrieved oocyte number to infant birth weight, were recorded in Mexican women undergoing in vitro fertilization (n = 88). Multiple regression analysis sought BMI-adjusted and age-adjusted associations. Receiver operating characteristic analysis tested for discrimination between outcomes. RESULTS: Associations of FF and serum FA were markedly distinct. While various FF FA (C16:1, C18:0, C20:2, C20:3, arachidonic acid) were significantly and inversely associated only with retrieved oocyte number, selected serum FA were associated with a broad range of pre-fertilization and post-fertilization parameters. Associations of BMI and FF FA were complex, as arachidonic acid was inversely associated with both BMI and retrieved oocyte number, while oleic acid (OA) was directly associated with BMI and PBF. Ultrasound-assessed clinical pregnancy outcome (CP) was directly associated with serum OA but inversely with its trans isomer elaidic acid (EA) and with BMI. Compounded BMI, serum EA and OA discriminated CP well (AUC = 0.74). Whole blood DNA methylation was significantly associated with and a moderate predictor (AUC = 0.66) of percent fertilized oocytes. CONCLUSIONS: Overall FF FA pool composition rather than FA identity may impact oocyte production and cellular memory of FF FA is lost as the oocyte exits the follicular environment. The contrasting associations of BMI, FF OA and arachidonic acid suggest that the control of oocyte homeostasis by FF FA is uncoupled from BMI. Further studies are warranted to assess the potential of compounding BMI with serum EA and OA to predict CP.

An Extremely Low-Frequency Vortex Magnetic Field Modifies Protein Expression, Rearranges the Cytoskeleton, and Induces Apoptosis of a Human Neuroblastoma Cell Line.

Homogeneous extremely low-frequency electromagnetic fields (ELF-EMFs) alter biological phenomena, including the cell phenotype and proliferation rate. Heterogenous vortex magnetic fields (VMFs), a new approach of exposure to magnetic fields, induce systematic movements on charged biomolecules from target cells; however, the effect of VMFs on living systems remains uncertain. Here, we designed, constructed, and characterized an ELF-VMF-modified Rodin's coil to expose SH-SY5Y cells. Samples were analyzed by performing 2D-differential-gel electrophoresis, identified by MALDI-TOF/TOF, validated by western blotting, and characterized by confocal microscopy. A total of 106 protein spots were differentially expressed; 40 spots were downregulated and 66 were upregulated in the exposed cell proteome, compared to the control cell proteome. The identified spots are associated with cytoskeleton and cell viability proteins, and according to the protein-protein interaction network, a significant interaction among them was found. Our data revealed a decrease in cell survival associated with apoptotic cells without effects on the cell cycle, as well as evident changes in the cytoskeleton. We demonstrated that ELF-VMFs, at a specific frequency and exposure time, alter the cell proteome and structurally affect the target cells. This is the first report showing that VMF application might be a versatile system for testing different hypotheses in living systems, using appropriate exposure parameters.© 2022 Bioelectromagnetics Society.

Oocyte maturation arrest produced by TUBB8 mutations: impact of genetic disorders in infertility treatment.

Oocyte maturation defect is a challenging situation in the management of infertility, the etiology may be related to endocrine causes, protocols used in ovarian stimulation, oocyte intrinsic defects or procedures in embryology laboratory. We report three Mexican females in treatment for primary infertility with non-mature oocytes after ovary stimulation and oocyte capture in whom a genetic diagnosis of TUBB8-oocyte maturation defect was revealed by exome sequencing. Two couples achieved pregnancies though oocyte donation after establishing the genetic etiology. Our results expand the role of TUBB8-disorders in patients of non-Asian ethnicity. Oocyte maturation defects of monogenic origin are a growing group of disorders that endocrinologists and reproductive medicine specialists should be aware in order to provide referral to genetics for establish a correct and opportune diagnosis.

Natural-cycle in vitro fertilization (IVF) combined with in vitro maturation in infertile patients with polycystic ovarian syndrome (PCOS) requiring IVF.

OBJECTIVE: In this study, we report an experience of 59 natural-cycle IVF combined with in vitro oocyte maturation (IVF/M) cycles in patients with PCOS requiring IVF recruited based on limitations to afford a conventional IVF treatment in a 9-years period. Results of IVF/M were compared with 164 cycles of IVF in PCOS patients. MATERIAL AND METHODS: In IVF/M cycles only hCG priming was used before oocyte recovery, with in vitro maturation of immature oocytes in a commercial medium. In conventional IVF group, recombinant FSH (rFSH) and GnRH agonist/antagonist for ovarian stimulation were used. In both groups, fertilization was achieved by intracytoplasmic sperm injection (ICSI) of mature oocytes and fresh embryos transferred at day 2 or day 3. RESULTS: In all IVF/M cycles oocytes and transferable quality embryos were obtained, only in 6 IVF/M cycles mature oocytes were obtained at oocyte capture day. Clinical pregnancy rate per cycle was 39.0% vs 53.6% (p = 0.0682) and delivery rate per cycle was 30.5% vs 42.6% (p = 0.1209) in IVF/M and conventional IVF respectively. Patients with ovarian hyperstimulation syndrome (OHSS) were 0% in IVF/M vs 6.7% in conventional IVF (p = 0.0399). CONCLUSION: Our experience in a private clinic in Mexico suggests that IVF/M can be a useful initial strategy to treat PCOS patients requiring IVF with comparable delivery rates to conventional IVF and a decreased risk of ovary hyperstimulation. IVF/M may be indicated to patients with limited resources paying without insurance for their infertility treatment.

[Impact of laser-assisted hatching (quarter technique) in poor prognosis patients]. / Impacto de la eclosión asistida con láser (técnica en cuartos) en pacientes con pobre pronóstico reproductivo.

BACKGROUND: Poor implantation rates continue to be the determinant factor for results in assisted reproductive techniques; many factors are thought to be involved including embryo quality, endometrial receptivity and embryo transfer. Assisted hatching has been proposed as a technique to improve implantation rates in selected groups of patients, especially with poor prognosis. OBJECTIVE: To evaluate the impact of the laser assisted hatching performed with quarter technique in patients with poor prognosis. MATERIAL AND METHODS: Prospective, controlled and randomized clinical study. The study group included patients with poor prognosis: maternal age ≥ 38 years, basal FSH ≥ 12.0 mUl/mL, two or more previous FIV/ ICSI failures. The control group don't received neither assisted reproductive techniques. RESULTS: We registered 303 cycles: n=1 54 in study group (laser assisted hatching) and n = 149 in control group. Clinical pregnancy (40.1 vs 19.7%) and implantation (17.5 vs 8.3%) rates were significant higher in laser assisted hatching group, there were not significant differences between multiple pregnancy (13.11 vs 10%) and miscarriage (14.7 vs 17.2) rates. CONCLUSION: Laser assisted hatching with quarter technique improves pregnancy and implantation rates in poor prognosis patients.

MRP1 overexpression determines poor prognosis in prospectively treated patients with localized high-risk soft tissue sarcoma of limbs and trunk wall: an ISG/GEIS study.

Patients with localized high-risk soft tissue sarcomas (STS) of the limbs and trunk wall still have a considerable metastatic recurrence rate of more than 50%, in spite of adjuvant chemotherapy. This drug-ceiling effect of chemotherapy in sarcoma setting could be explained, at least partially, by multidrug resistance (MDR) mechanisms. The aim of this study was to ascertain whether mRNA and protein expression of ABCB1 (P-glycoprotein), ABCC1 (MRP1), and GSTA1 (glutathione S-transferase pi) was prognostic in localized high-risk STS. Immunohistochemistry and reverse transcriptase-PCR studies were performed from biopsies at the time of diagnosis. Patients of this series were prospectively enrolled into a phase III trial that compared three versus five cycles of epirubicin plus ifosfamide. The series of 102 patients found 41 events of recurrence and 37 of death with a median follow-up of 68 months. In univariate analysis, variables with a statistically significant relationship with relapse-free survival (RFS) were: MRP1 expression (5-year RFS rate of 23% in positive cases and 63% in negative cases, P = 0.029), histology (5-year RFS rate of 74% in undifferentiated pleomorphic sarcoma and 43% in synovial sarcoma, P = 0.028), and ABCC1 expression (5-year RFS rate of 33% in overexpression and 65% in downregulation, P = 0.012). Combined ABCC1/MRP1 was the only independent prognostic factor for both RFS (HR = 2.704, P = 0.005) and overall survival (HR = 2.208, P = 0.029). ABCC1/MRP1 expression shows robust prognostic relevance in patients with localized high-risk STS treated with anthracycline-based chemotherapy, which is the standard front line treatment in STS. This finding deserves attention as it points to a new targetable protein in STS.

Y chromosome microdeletions in Mexican males of couples with idiopathic recurrent pregnancy loss.

AIM: To analyze the presence of Y chromosome microdeletions in males of Mexican couples with idiopathic recurrent pregnancy losses (RPL). METHODS: Seventy-one males from couples with RPL and 66 fertile males as controls were studied. DNA was isolated from peripheral lymphocytes and used to run multiplex polymerase chain reactions. Regions AZFa (sY84, sY86), AZFb (sY127, sY134) and AZFc (sY254, sY255) of the Y chromosome were analyzed according to valid guidelines recommended by the European Academy of Andrology and the European Molecular Genetics Quality Network. Also, the sequence tagged sites (STSs): DYS262 (sY67), DYS220 (sY129), DYF85S1 (sY150), DYF86S1 (sY152) and DYF87S1 (sY153) were included in order to analyze STSs previously reported as deleted. A power analysis to support our simple size was performed. RESULTS: Results show an absence of Y chromosome microdeletions in males of couples with RPL and controls with an acceptable statistical power. CONCLUSION: The study did not show an association of recurrent pregnancy loss and Y chromosome microdeletions in Mexican male partners. Based on the results, the study of Y chromosome microdeletions in couples with RPL is not considered clinically relevant.

Estudio de los mecanismos de transporte iónico involucrados en la secreción en túbulos de malpighi de chipo (Rhodnius prolixus) / Study of the mechanism of ionic transport involved on the secretion of malpighian tubes of chipos (Rhodnius prolixus)

Se midió la tasa de secreción y los cambios en la concentración interna de sodio y protones de los túbulos demalpighi (UMT) en Rhodnius prolixus bajo estimulación con 5-OH-Triptamina. Para medir secreción utilizamos un sistema de doble perfusión para tener acceso por separado a las membranas basolateral y/o apical de la célula. Los flujos iónicos se midieron por microfluorometría. Se aplicaron hasta trece agentes farmacológicos: Ouabaina, Bafilomicina A1, Furosamida, Bumetanida, DIOA, Probenecina, SITS, Acetazolamida, Amilorida, DPC, BaCl2, pCMBS y DTT. Estos agentes son bloqueadores conocidos de diferentes funciones de transporte de iones, conocidos como ATPasas, co- y/o contratransportadores asi como de canales iónicos y de agua. La asunción básica es que los cambios en los movimientos de agua reflejan los cambios en los mecanismos del transporte de iones que se localizan de la siguiente manera: (i) en la membrana basolateral de la célula, los principales son un cotransportador de Na+-K+-2Cl– y el intercambiador de Cl--HCO- 3; la ATPasa de Na+- K+ y la ATPasa de Na+ y los canales de Cl- y de agua (Rp-MIP) son de importancia intermedia, mientras que los canales de K+ son menos importantes: (ii) en la membrana celular apical, la mayor importancia la tiene un cotransportador de K+-Cl-, que ha sido localizado por primera vez, una ATPasa de H+ tipo V, y un intercambiador de Na+-H+; el intercambiador urato-anión y los canales de K+ son importantes mientras que los canales de Clno lo son. En este trabajo presentamos un modelo actualizado que explica la secreción del UMT.

We have measured fluid secretion rate and intracelular concentration of sodium and protons, in Rhodnius prolixusUMT stimulated to secrete with 5-OH-tryptamine. We used double perfusions in order to have access separately to the basolateral and to the apical cell membranes. The ionic fluxes has been measured using microfluorimetry. 13 pharmacological agents were applied: Ouabain, Bafilomycin A1, Furosemide, Bumetanide, DIOA, Probenecid, SITS, Acetazolamide, Amiloride, DPC, BaCl2, pCMBS and DTT. These agents are known to block different ion transport functions, namely ATPases, co- and/or counter- transporters and ionic and water channels. The basic assumption is that water movement changes reflect changes on ion transport mechanisms which we localize as follows: (i) At the basolateral cell membrane: a Na+ - K+ - 2Cl- cotransport; a Cl- - HCO3 - exchange; the Na+ - K+ - ATPase, Cl- channels and Rp-MIP water channels are fundamental; K+ channels play a lesser role. (ii) At the apical cell membrane, a K+ - Cl- cotransport that is located for the first time; a V - H+ - ATPase; an Na+– H+ exchange; an urate - anion exchange and K+ channels are important, while Cl- channels are not. The present experiments allow us to build a tentative model for the function of the UMT cell, which includes a paracellular pathway for fluid flow. In this work we propose a new actualized model to explain the UMT secretion.

[Y chromosome micro-deletion identification in infertile males]. / Identificación de microdeleciones en el cromosoma Y en pacientes con infertilidad masculina.

UNLABELLED: Identifying the genetic causes of male infertility is very important, considering they account for 30-50% of reproductive problems among couples. Genetic abnormalities, among which Y chromosome microdeletions are found, are commonly detected in patients with non-obstructive azoospermia (0-4.3%). Most of these patients are eligible for intracytoplasmic sperm injection (ICSI) and this genetic defect can be inherited by male children. OBJECTIVE: Determining the prevalence of microdeletions in the Y chromosome in a group of Mexican patients presenting azoospermia and oligospermia, under treatment in the Infertility Clinics. MATERIALS AND METHODS: This study included 52 infertile men (cases): 36 with non-obstructive azoospermia, and 16 with oligospermia; and 50 men (controls) whose fertility had been validated. The genomic DNA of each individual was obtained from his EDTA and heparin treated blood, and the corresponding karyotype determined. The karyotype was analyzed using G banding techniques. Eighteen markers (STS) corresponding to the chromosome Y long arm (AZFa, b, c, and d zones) were amplified in each DNA sample in all cases--azoospermic, oligospermic and controls--using the PCR method. RESULTS: No chromosomal alterations were detected in the patients, and no Y chromosome microdeletions were detected in control cases. Five azoospermic patients (13.9%) presented microdeletions corresponding to the AZFb, c, and d zones, while no microdeletions were found in oligospermic patients. The frequency of microdeletions found in this study is very similar to that reported for other populations. CONCLUSIONS: This research not only reports the frequency of microdeletions in the Y chromosome in our population, but also contributes to the integration of a DNA bank for patients with idiopathic male infertility, which will be of great use in the search for the causes of this affection.

Nitric oxide induces resensitization of P2Y nucleotide receptors in cultured rat mesangial cells.

Receptor desensitization of G protein-coupled receptors (GPCRs), which occurs during short-term (seconds to minutes) exposure of cells to agonists, is mediated by phosphorylation and receptor endocytosis. Recycling of the receptors is a requisite for resensitization of the response. The mechanisms that attenuate signaling by GPCRs are of considerable importance to regulation of intercellular signaling and maintenance of their ability to respond to agonists over time. This study evaluates the effect of nitric oxide (NO) on P2Y nucleotide receptor resensitization in cultured rat glomerular mesangial cells. The NO production in cultured mesangial cells was measured by using confocal microscopy and the fluorescence NO indicator 4,5-diaminofluorescein diacetate (DAF-2 DA). L-arginine increased and Nomega-nitro-L-arginine methyl ester (L-NAME) decreased NO production significantly (P < 0.05). Calcium responses to ATP were measured with fura-2 and imaging techniques. Repeated stimulation with ATP results in receptor desensitization that is characterized by lower calcium peak amplitude. Desensitization was induced by challenging mesangial cells with four consecutive 2-min pulses of ATP (0.1 mM) separated by 4.5-min control perfusions. Intracellular calcium concentration ([Ca2+]i) increase evoked by second, third, and fourth ATP challenges were about 40%, 26%, and 18% of the first one. The NO precursor, L-arginine (10 mM), and the NO donors, spermine-NONOate (500 microM) and sodium nitroprusside (SNP) (1 mM), were added before and during a fourth ATP challenge. Spermine-NONOate and L-arginine induced a recovery of the [Ca2+]i response to the fourth ATP challenge (P < 0.01 and 0.05, respectively). The NO synthase inhibitor, L-NAME (5 mM), applied along with ATP, was shown to enhance desensitization. 1H-(1,2,4)oxadiazolo(4,3-alpha)quinoxalin-1-one (ODQ, 30 microM), an inhibitor of guanylate cyclase, was used along with L-arginine, SNP, or spermine-NONOate. There was no significant difference with or without ODQ. Neither ODQ nor 8-Br-cGMP, an analog of cGMP, at different concentrations showed effects on ATP-stimulated [Ca2+]i. There was no elevation of [Ca2+]i when the cells were challenged by different concentrations (1 microM, 100 microM, 1 mM, 20 mM, and 30 mM) of caffeine, caffeine plus ATP (0.1 mM), and 4-chloro-3-ethylphenol (100 microM, 500 microM, and 1 mM), a new agonist of ryanodine receptors. The results indicate that NO can increase the P2Y receptor resensitization in rat glomerular mesangial cells by acting through a cGMP-independent pathway. No evidence was found for the existence of ryanodine-sensitive intracellular calcium stores in rat mesangial cells.

Efecto de la reducción de peso en la condición clínica y hormonal de mujeres anovulatorias obesas / Neight reduction effect in clinical and hormonal condition of obese anovulatory women

La obesidad se asocia a una gran variedad de trastornos endocrinos hipotálamo-hipófisio-ováricos que pueden llevar a una anovulación persistente. La reducción de peso puede mejorar el perfil hormonal y por lo tanto recuperar la función ovulatoria. El objetivo del presente estudio fue valorar el efecto de la reducción de peso en la condición clíonica y hormonal de mujeres anovulatorias obesas en el Instituto de Medicina Reproductiva del Bajío en el Hospital Aranda de la Parra de la ciudad de León, Guanajuato. Se analizaron un total de 30 pacientes entre 18 y 35 años de edad con obesidad, anovulación crónica y sin patología tiroidea. Previo y posterior a un tratamiento de reducción de peso, con pérdida de al menos 5 por ciento de peso inicial, se analizaron hormona luteinizante (LH), hormona folículo estimulante (FSH), estradiol, prolactina, testosterona, dihidroepiandrostendiona-sulfato (DEA-S), curva de tolerancia oral a la glucosa y progesterona en día 21, peso, IMC, relación cintura/cadera y porcentaje de grasa por suma de pliegues. La pérdida media de peso fue de 9.5 ñ 4.3 kg. lo que representa una pérdida de peso de 10.96 por ciento con respecto al inicial. Se presentó ovulación espontánea en 26 pacientes (86.6 por ciento). Hubo una reducción significativa en los niveles basales de LH, estradiol, testoterona, DHEA-S, así como un aumento en los niveles de progesterona. De 12 pacientes con curva de tolerancia a la glucosa alterada, nueve (75 por ciento) observaron mejoría la final del tratamiento. Los resultados obtenidos demuestran que la disminución de peso y de porcentaje corporal de grasa puede mejorar el perfil hormonal y la función ovulatoria de pacientes anovulatorias obesas por lo que estas mujeres deberían someterse a un tratamiento de reducción de peso antes de comenzar con inductores de ovulación

Embarazo gemelar mediante la técnica de donación de ovocitos en una mujer con síndrome de Turner / Twin pregnancy by oocyte donation technique in a woman with Turner's syndrome

Es extremadamente raro que ocurra un embarazo espontáneo en pacientes con síndrome de Turner, y en caso de presentarse este correrá un gran riesgo de pérdidas fetales y malformaciones congénitas en el recién nacido, especialmente del cromosoma sexual. La donación de ovocitos es actualmente la técnica de elección para solucionar los deseos reproductivos de estas mujeres. Se presenta el caso de una mujer con síndrome de Turner, cariotipo 45, XO, a la cual se le realiza donación de ovocitos y transferencia intrauterina de embriones, logrando un embarazo gemelar, el cual transcurrió sin complicaciones, se efectuó una cesárea obteniendo dos productos completamente sanos