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1.
Reprod Domest Anim ; 59(3): e14548, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38459830

RESUMO

The in vivo fertilization process occurs in the presence of follicular fluid (FF). The aim of this study was to evaluate the effect of in vitro fertilization medium supplementation with 5% or 10% bovine follicular fluid (BFF) on the production of in vitro bovine embryos. FF was collected from ovarian follicles with a diameter of 8-10 mm, and cumulus-oocyte complexes (COCs) were co-incubated with sperm for 24 h in the commercial medium BotuFIV® (BotuPharma©), being distributed among the experimental groups: oocytes fertilized in a control medium; oocytes fertilized in a medium supplemented with 5% BFF; and oocytes fertilized in a medium supplemented with 10% BFF. After fertilization, the zygotes were cultured in vitro for 8 days. Embryo development was assessed through cleavage rates (day 2) and blastocyst formation rates (day 8). The relative expression of the genes OCT4, IFNT2, BAX, HSP70 and SOD2 was measured using the real-time polymerase chain reaction method. There was no difference (p > .05) among the different experimental groups in terms of cleavage rates and blastocyst formation rates. Regarding the gene expression results, only the blastocysts from oocytes fertilized with 10% BFF showed significantly lower expression of IFNT2 (p = .003) and SOD2 (p = .01) genes compared to blastocysts from oocytes fertilized in control medium alone, while there was no difference between blastocyst from oocytes fertilized in control medium and the ones from oocytes fertilized with 5% BFF. In addition to this, the blastocysts from oocytes fertilized with 5% BFF showed significantly reduced levels of expression of the heat shock protein HSP70 (p < .001) and the pro-apoptotic protein BAX (p = .015) compared to blastocysts from oocytes fertilized with control medium. This may indicate that lower supplementation of BFF to the IVF medium creates a more suitable environment for fertilization and is less stressful for the zygote.


Assuntos
Fertilização in vitro , Líquido Folicular , Feminino , Masculino , Bovinos , Animais , Proteína X Associada a bcl-2/genética , Proteína X Associada a bcl-2/metabolismo , Fertilização in vitro/veterinária , Sêmen , Oócitos , Desenvolvimento Embrionário , Blastocisto/metabolismo , Proteínas de Choque Térmico HSP70/genética , Fertilização
2.
JBRA Assist Reprod ; 28(2): 263-268, 2024 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-38381778

RESUMO

OBJECTIVE: The objective of this research is to investigate the association between the concentrations of soluble human leukocyte G antigen (sHLA-G) in the follicular fluid (FF) in infertile patients with peritoneal endometriosis submitted to in vitro fertilization. METHODS: We performed a cross-sectional study, including ninety-six women undergoing in vitro fertilization (IVF) ageing ≤ 40 years. Infertile patients were classified into two groups: with endometriosis diagnosed by laparoscopy and without endometriosis due to tubal factor. ELISA measured soluble HLA-G in the FF of a pool of punctured (more than 17mm) follicles from women with endometriosis and without endometriosis who were subjected to ovulation induction for IVF. Embryos obtained after fertilization were classified according to the graduated embryo score (GES). RESULTS: Groups were comparables in terms of age, the number of follicles, AMH, FSH and all included reproductive outcomes. There was no association between sHLA-G concentrations and the average score of the generated embryos (p>0.05). Measurement of sHLA-G in the follicle fluid in women with endometriosis and without endometriosis (tubal factor) showed no significant difference (p>0.05). We also compared sHLA-G per follicle and per embryo, which were not different between both groups (p>0.05). CONCLUSIONS: Patients with peritoneal endometriosis submitted to IVF did not demonstrate an altered sHLA-G in the follicular fluid compared to the follicular fluid sHLA-G concentration in tubal factor patients. Also, this molecule was not linked to any other reproductive outcome.


Assuntos
Endometriose , Fertilização in vitro , Líquido Folicular , Antígenos HLA-G , Infertilidade Feminina , Humanos , Feminino , Endometriose/metabolismo , Líquido Folicular/metabolismo , Líquido Folicular/química , Adulto , Antígenos HLA-G/metabolismo , Antígenos HLA-G/análise , Estudos Transversais , Infertilidade Feminina/metabolismo , Gravidez
3.
JBRA Assist Reprod ; 2023 Sep 28.
Artigo em Inglês | MEDLINE | ID: mdl-37768816

RESUMO

OBJECTIVE: To assess whether follicular fluid (FF) from infertile women with endometriosis in advanced stages [moderate/severe (EIII/IV) without or with endometrioma (Endometrioma)] induce more oocyte damages than in early stages (minimal/mild: EI/II); and whether supplementation with L-carnitine (LC) and omega 3 (n3) can prevent these oocyte damages. METHODS: Experimental study using bovine oocytes (obtained of ovaries from slaughterhouse), and human FF (samples were obtained during oocyte recovery for ICSI). Bovine oocytes were submitted to in vitro maturation (IVM) divided into 9 groups: no FF(No-FF), with 1% FF from infertile women without endometriosis (FFC), with EI/II, EIII/IV and Endometrioma, and with (or not) LC+n3 addition. After IVM, oocytes were fluorescently labelled and visualized by confocal microscopy to analyze chromosomes and spindle. RESULTS: FF from endometriosis decreased rate of normal MII (spindle assembly and chromosome alignment) compared to No-FF (87.2%) and FFC (87.2%). FFEIII/IV (80.7%) and FFEndometrioma (69.3%) decreased total MII rate compared to No-FF (91.9%) and FFC (89.2%), and FFEndometrioma had lower total MII rate compared to other groups. LC+n3 increased MII rate in the FFEIII/IV (80.7% vs. 90.8%) and the Endometrioma (69.3% vs. 86.4%), and it prevented damages in spindle and chromosomes in MII oocytes in the FFEI/II group (62.2% vs. 84.5%) and the FFEIII/IV group (70.2% vs. 84.1%). CONCLUSIONS: FF of endometriosis damaged the meiotic spindle of bovine MII oocytes. EIII/IV led to impaired nuclear maturation; FF from women with endometrioma had further negative impact in oocyte maturation. LC+n3 completely prevented the effects of FF from women with endometriosis on oocyte.

4.
Reprod Domest Anim ; 58(12): 1662-1671, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-37743826

RESUMO

In the postpartum period, there is an increase in non-esterified fatty acids (NEFA) in both serum and follicular fluid (FF) of cattle. The increase in fatty acid concentration results in increased production of reactive oxygen species (ROS) that can compromise bovine fertility. The objectives of this study were to characterize the lipid profile found in the FF of cows experiencing induced negative energy balance (NEB) and to evaluate the effect of α-tocopherol in the prevention of oxidative stress in the serum and FF of cows. Twenty-nine beef cows were divided into groups: (1) control; (2) Fasting for 24 days; and (3) Fasting + VitE. Between D0 and D4 blood samples were taken to assess concentrations of NEFA, ROS production, total antioxidant capacity (FRAP), lipid peroxidation, and α-tocopherol (vitamin E). On D4, follicular aspiration was performed for analysis of FF from the dominant follicle. Our results demonstrate that fasting was effective in causing increased fat mobilization in animals. The increase in serum concentration of C18:1c9 was reflected in the FF of fasting cows. Serum α-tocopherol concentration was higher in the control and Fasting + VitE groups compared to the Fasting group. In FF, there was an increase of α-tocopherol in the Fasting + VitE group in comparison to Fasting cows. There was an increase in ROS production in the serum of fasting cows. ROS production in FF was higher in the Fasting compared to the Fasting + VitE group. Vitamin E has beneficial effects in reducing ROS production in the dominant follicle of cows in NEB.


Assuntos
Ácidos Graxos não Esterificados , Vitamina E , Feminino , Bovinos , Animais , Espécies Reativas de Oxigênio , Vitamina E/farmacologia , Lactação/metabolismo , alfa-Tocoferol/farmacologia
5.
Expert Rev Mol Diagn ; 23(2): 123-132, 2023 02.
Artigo em Inglês | MEDLINE | ID: mdl-36856088

RESUMO

INTRODUCTION: Polycystic ovary syndrome (PCOS) is a condition that affects approximately 13% of reproductive age women and is characterized by androgen excess, menstrual irregularity and altered ovarian morphology. PCOS presents a complex etiology and pathophysiology, which still requires a detailed investigation of biochemical signatures to identify the molecules and mechanisms that govern it. AREAS COVERED: This narrative review summarizes the main molecular alterations found in the ovarian follicular fluid, endometrium and placenta of women with PCOS, and the genotypes potentially associated with the outcome of infertility treatments in PCOS. EXPERT OPINION: PCOS is associated with multiple alterations in growth factors, sex steroid hormones, reactive oxygen species, proinflammatory cytokines and adipokines, which contribute to follicle arrest/ anovulation or suboptimal corpus luteum function, and ultimately to menstrual irregularity and hyperandrogenic symptoms. A panel of PCOS biomarkers should include, besides ovarian products, markers of adipose tissue function, insulin resistance, vascular health, and low-grade chronic inflammation. The effects of ovarian stimulation drugs on infertile women with PCOS are likely to be modified by genetic factors, but the available evidence is heterogeneous; therefore, future studies should evaluate standard treatments and pre-specified outcomes of interest to provide more conclusive answers.


Assuntos
Infertilidade Feminina , Síndrome do Ovário Policístico , Gravidez , Feminino , Humanos , Síndrome do Ovário Policístico/complicações , Síndrome do Ovário Policístico/diagnóstico , Infertilidade Feminina/diagnóstico , Infertilidade Feminina/etiologia , Adipocinas , Citocinas , Genótipo
6.
Animals (Basel) ; 13(6)2023 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-36978604

RESUMO

This study investigated the seasonal variations in mRNA expression of FSH (Fshr), LH (Lhr) receptors, melatonin (Mt1 and Mt2) receptors, melatonin-synthetizing enzymes (Asmt and Aanat) and melatonin concentration in developing follicles from mares raised in natural photoperiods. For one year, ultrasonographic follicular aspiration procedures were performed monthly, and small (<20 mm), medium (20 to 35 mm) and large (>35 mm) follicles were recovered from five mares. One day before monthly sample collections, an exploratory ultrasonography conducted to record the number and the size of all follicles larger than 15 mm. The total number of large follicles were higher during the spring/summer (8.2 ± 1.9) than during autumn/winter (3.0 ± 0.5). Compared to autumn/winter seasons, there was an increase of Fshr and Aanat mRNA expressions in small, medium and large follicles, an increase of Lhr and Asmt mRNA expressions in medium and large follicles and an increase of Mt1 and Mt2 mRNA expressions in small and large follicles during spring/summer. The melatonin levels in follicular fluid were also higher during the spring/summer seasons. The present data show that melatonin locally upregulates the mRNA expression of Mt1 and Mt2 receptors and melatonin-forming enzymes in mare developing follicles during reproductive seasons.

7.
JBRA Assist Reprod ; 26(2): 288-298, 2022 04 17.
Artigo em Inglês | MEDLINE | ID: mdl-34786903

RESUMO

OBJECTIVE: Human follicular fluid (FF) contains different cell populations including mesenchymal stem cells. Studies tried to improve their differentiation to oocyte and use them in infertility treatments. Using an antioxidant may improve the quality of these cells. The present study investigated the effects of different doses of melatonin on FF-derived cells grown to oocyte-like cells (OLC). METHODS: Cell viability (MTT assay), flow cytometry, and ICC staining were utilized to evaluate CD105 and CD34 expression; colony forming unit assay (CFU-F) capability, qRT-PCR were used to investigate ZP1, ZP2, ZP3, GDF9, and SCP3 expression. AMH, Estradiol and Progesterone levels in the supernatant were measured. Morphological characteristics of fibroblast-like cells changing to a round shape were seen specifically in the group treated with melatonin 10-7M after 2 weeks. RESULTS: There was no difference between control and treatment groups for MTT and CFU assays. ICC staining was positive for CD105 marker and negative for CD34 hematopoietic stem cell marker. qRT-PCR results indicated that ZP1, ZP2, GDF9, and SCP3 expression increased in the group treated with melatonin 10-7M in Week 2, while ZP3 decreased in this group. Progesterone and AMH were detected in differentiation medium. CONCLUSIONS: Melatonin may improve in vitro formation of OLCs.


Assuntos
Líquido Folicular , Melatonina , Diferenciação Celular , Feminino , Líquido Folicular/metabolismo , Humanos , Melatonina/farmacologia , Oócitos , Progesterona/farmacologia
8.
Biochim Biophys Acta Mol Basis Dis ; 1868(1): 166295, 2022 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-34718118

RESUMO

Several organs, such as the heart, breasts, intestine, testes, and ovaries, have been reported to be target tissues of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. To date, no studies have demonstrated SARS-CoV-2 infection in the female reproductive system. In the present study, we investigated the effects of SARS-CoV-2 infection on ovarian function by comparing follicular fluid (FF) from control and recovered coronavirus disease 2019 (COVID-19) patients and by evaluating the influence of these FF on human endothelial and non-luteinized granulosa cell cultures. Our results showed that most FFs (91.3%) from screened post COVID-19 patients were positive for IgG antibodies against SARS-CoV-2. Additionally, patients with higher levels of IgG against SARS-CoV-2 had lower numbers of retrieved oocytes. While VEGF and IL-1ß were significantly lower in post COVID-19 FF, IL-10 did not differ from that in control FF. Moreover, in COV434 cells stimulated with FF from post COVID-19 patients, steroidogenic acute regulatory protein (StAR), estrogen-receptor ß (Erß), and vascular endothelial growth factor (VEGF) expression were significantly decreased, whereas estrogen-receptor α (ERα) and 3ß-hydroxysteroid dehydrogenase (3ß-HSD) did not change. In endothelial cells stimulated with post COVID-19 FF, we observed a decrease in cell migration without changes in protein expression of certain angiogenic factors. Both cell types showed a significantly higher γH2AX expression when exposed to post COVID-19 FF. In conclusion, our results describe for the first time that the SARS-CoV-2 infection adversely affects the follicular microenvironment, thus dysregulating ovarian function.


Assuntos
COVID-19/metabolismo , COVID-19/virologia , Interações Hospedeiro-Patógeno , Ovário/metabolismo , Técnicas de Reprodução Assistida , SARS-CoV-2 , Adulto , Anticorpos Antivirais/imunologia , Biomarcadores , COVID-19/imunologia , Células Cultivadas , Citocinas/metabolismo , Ensaio de Imunoadsorção Enzimática , Feminino , Fertilidade , Líquido Folicular/metabolismo , Células da Granulosa/metabolismo , Interações Hospedeiro-Patógeno/imunologia , Humanos , Imunoglobulina G/imunologia , Oócitos/metabolismo , Adulto Jovem
9.
Int J Mol Sci ; 22(7)2021 Mar 26.
Artigo em Inglês | MEDLINE | ID: mdl-33810351

RESUMO

Lipids are a potential reservoir of energy for initial embryonic development before activation of the embryonic genome and are involved in plasma membrane biosynthesis. Excessive lipid droplet formation is detrimental to cryotolerance and is related to alterations in mitochondrial function, which likely affects lipid metabolism. Increased lipid accumulation in in vitro produced embryos is a consequence of the stress during in vitro embryonic development process. There are several open questions concerning embryo lipid metabolism and developmental potential. Oocyte maturation and embryo development in vivo and in vitro may vary if the donors are subjected to any type of stress before follicle puncture because crucial changes in oocyte/embryonic metabolism occur in response to stress. However, little is known about lipid metabolism under additional stress (such as heat stress). Therefore, in this review, we aimed to update the information regarding the energy metabolism of oocytes and early bovine embryos exhibiting developmental competence, focusing on lipid metabolic pathways observed under in vivo, in vitro, and stress conditions.


Assuntos
Técnicas de Maturação in Vitro de Oócitos , Metabolismo dos Lipídeos , Oócitos/metabolismo , Oogênese/genética , Animais , Blastocisto/metabolismo , Bovinos , Criopreservação , Células do Cúmulo/citologia , Células do Cúmulo/metabolismo , Feminino , Temperatura Alta , Humanos , Lipídeos/química , Camundongos , Mitocôndrias/metabolismo , Oócitos/citologia , Folículo Ovariano/metabolismo , Especificidade da Espécie , Suínos
10.
Rev. bras. reprod. anim ; 45(4): 376-388, out.-dez. 2021. graf, ilus
Artigo em Inglês | VETINDEX | ID: biblio-1492684

RESUMO

O desenvolvimento de estudos genéticos e de microdispositivos biológicos tem proporcionado a ampliação do conhecimento sobre os complexos eventos que envolvem a reprodução animal. O desafio ainda é imensurável, mas a criação e surgimentos de novas perspectivas para a pesquisa básica tem-se feito presente. Neste trabalho revisamos de maneira suscinta algumas abordagens recentes, utilizadas pela pesquisa básica, sobretudo com o objetivo de lançar luz sobre o desenvolvimento folicular e oocitário. Dessa forma, essa revisão pretende fornecer uma visão geral do uso das tecnologias ômicas e sistema de microfluídica como auxiliadores na compreensão da foliculogênese. Adicionalmente serão apresentadas particularidades inerentes à fisiologia da gametogênese, que incluem ação de microorganismos e mitocôndrias, além do importante papel da comunicação intercelular através das vesículas extracelulares.


The development of genetic studies and biological microdevices has expanded knowledge about the complex events involving animal reproduction. The challenge is still immeasurable, but the creation and emergence of new perspectives for basic research have been present. This paper briefly reviews some recent approaches used in basic research, mainly to shed light on follicular and oocyte development. Thus, this review intends to provide an overview of the use of omics technologies and microfluidics systems as aids in understanding folliculogenesis. Also, it will present particulars inherent in the physiology of gametogenesis, which include microorganisms and mitochondria, in addition to the important role of intercellular communication through extracellular vesicles.


Assuntos
Animais , Bioengenharia , Fenômenos Genéticos , Folículo Ovariano/crescimento & desenvolvimento , Microbiota , Técnicas Analíticas Microfluídicas
11.
JBRA Assist Reprod ; 24(3): 245-249, 2020 07 14.
Artigo em Inglês | MEDLINE | ID: mdl-32155015

RESUMO

OBJECTIVE: Our aim was to analyze the role of thyroid hormones in follicular fluid (FF) in relation to the number of oocytes retrieved in women recruited for an assisted fertilization procedure. METHODS: Retrospective cohort study of 51 women 37.5±3.3 years, range 29-42, evaluated after a controlled ovarian stimulation protocol in a University Hospital. FF was sampled by transvaginal ultrasound-guided aspiration after ovarian hyperstimulation and we measured T3 (T3f), T4 (T4f), TSH (TSHf) and free T4 (T4ff). The oocyte maturation rate was calculated as: Number of metaphase II oocytes/Number of oocytes retrieved x 100. Statistical analysis was performed using the SPSS-19 software. RESULTS: Hormone levels in FF were: TSHf 1.3µIU/ml (0.4 - 2.7), T3f: 1.52±0.46 nmol/L, T4f 88.8±30.9nmol/L and T4ff: 15.44±2.57pmol/L. The number of oocytes recovered was dependent onT4f following the equation: Log (oocyte) = 0.379+0.042*T4f (r:0.352, p=0.012). After a logistic regression model analysis, T3f showed a tendency to be associated with the OMR: OR (95 % CI)= 0.977 (0.954 to 1.001), p=0.057. CONCLUSIONS: The correlation found between thyroid hormones and the number of oocytes retrieved suggests an interaction between thyroid and gonadal axes in relation to follicular development.


Assuntos
Líquido Folicular/metabolismo , Recuperação de Oócitos , Oócitos/metabolismo , Indução da Ovulação/métodos , Hormônios Tireóideos/metabolismo , Adulto , Feminino , Humanos
12.
J Assist Reprod Genet ; 37(1): 53-61, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31823133

RESUMO

Even the strictest laboratories and clinics are prone to the occurrence of microbial contamination. In the case of in vitro fertilization (IVF) research and practice facilities, the number of possible sources is particularly vast. In addition to ambient air, personnel, and non-sterilized materials, follicular fluid and semen from patients are a very common gateway for a diverse range of bacteria and fungi into embryo cultures. Even so, reports of contamination cases are rare, what leads many clinics to see the issue as a negligible risk. Microbiological contamination may result in the demise of the patient's embryos, leading to additional costs to both the patient and the clinics. Regardless of financial loss, emotional costs, and stress levels during IVF are highly distressing. Other worrisome consequences include DNA fragmentation, poor-quality embryos, early pregnancy loss or preterm birth, and possible long-term damages that need further investigation. In this review, we aimed to shed a light on the issue that we consider largely underestimated and to be the underlying cause of poor IVF outcomes in many cases. We also discuss the composition of the microbiome and how its interaction with the reproductive tract of IVF-seeking patients might influence their outcomes. In conclusion, we urge clinics to more rigorously identify, register, and report contamination occurrences, and highlight the role of the study of the microbiome to improve overall results and safety of assisted reproduction.


Assuntos
Bactérias/isolamento & purificação , Infecções Bacterianas/economia , Infecções Bacterianas/epidemiologia , Fertilização in vitro/economia , Fertilização in vitro/normas , Técnicas de Reprodução Assistida/economia , Infecções Bacterianas/microbiologia , Feminino , Humanos , Gravidez , Técnicas de Reprodução Assistida/normas
13.
Artigo em Inglês | MEDLINE | ID: mdl-31379738

RESUMO

Patients with poor ovarian response (POR) to exogenous gonadotropins stimulation for assisted reproductive technology (ART) have decreased circulating androgens during spontaneous cycles. The Patient-Oriented Strategies Encompassing Individualized Oocyte Number (POSEIDON) is a 4-tier stratification of women with POR to controlled ovarian stimulation (COH) based on age and biomarkers of ovarian reserve has been proposed to maximize the clinical management of this group for ART. The aim of the present study was to characterize the levels of androgens during COH in follicular fluid (FF) and serum in POSEIDON subgroups and compared them with women of normal ovarian response. Sixty nine consecutive patients undergoing ART were included and testosterone, androstenedione, dehydroepiandrosterone sulfate (DHEA-S), estradiol, sex hormone-binding globulin (SHBG), and insulin-like growth factor 1 (IGF-1) were measured in serum and FF collected at the time of oocyte pick-up. The number of retrieved oocytes was registered for each patient for their allocation to the respective POSEIDON subgroup. The control group comprised 19 women and the POSEIDON group 1 (age < 35, normal ovarian reserve biomarkers) n = 14, group 2 (age ≥ 35, normal ovarian reserve biomarkers) n = 8, group 3 (age < 35, poor ovarian reserve biomarkers) n = 6 and group 4 (age ≥ 35, poor ovarian reserve biomarkers) n = 22. Serum levels of total testosterone, androstenedione and DHEA-S were not different in group 1 vs. control but significantly decreased in group 3 vs. control. DHEA-S in FF was also significantly decreased in group 3 vs. control. In addition, serum testosterone was decreased in groups 2 and 4 vs. control; and serum androstenedione and estradiol were reduced in group 4 vs. control. No differences were observed for estradiol, SHBG and IGF-1 in FF. Finally, a high correlation between serum and FF DHEA-S was observed when data from samples of all groups were pooled. Group 1 did not show hypoandrogenemia however group 3 had low levels of all measured androgens in serum and DHEA-S in FF. Such differences might help to better characterize and/or improve the clinical management of women with POR according to their respective POSEIDON stratification.

14.
Zygote ; 27(5): 285-298, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31412964

RESUMO

Ovulation is considered an inflammatory, cytokine-mediated event. Cytokines, which are recognized as growth factors with immunoregulatory properties, are involved in many cellular processes at the ovarian level. In this sense, cytokines affect fertility and are involved in the development of different ovarian disorders such as bovine cystic ovarian disease (COD). Because it has been previously demonstrated that ovarian cells represent both sources and targets of cytokines, the aim of this study was to examine the expression of several cytokines, including IL-1ß, IL-1RA, IL-1RI, IL-1RII, IL-4 and IL-8, in ovarian follicular structures from cows with spontaneous COD. The protein expression of these cytokines was evaluated by immunohistochemistry. Additionally, IL-1ß, IL-4 and IL-8 concentrations in follicular fluid (FF) and serum were determined by enzyme-linked immunosorbent assay (ELISA). In granulosa and theca cells, IL-1RI, IL-1RII, IL-1RA and IL-4 expression levels were higher in cystic follicles than in the control dominant follicles. The serum and FF concentrations of IL-1ß and IL-4 showed no differences between groups, whereas IL-8 concentration was detected only in FF of cysts from cows with COD. The FF and serum concentrations of IL-1ß and IL-8 showed no significant differences, whereas IL-4 concentration was higher in FF than in serum in both the control and COD groups. These results evidenced an altered expression of cytokines in ovaries of cows with COD that could contribute to the pathogenesis of this disease.


Assuntos
Líquido Folicular/metabolismo , Interleucinas/metabolismo , Cistos Ovarianos/metabolismo , Cistos Ovarianos/patologia , Animais , Estudos de Casos e Controles , Bovinos , Doenças dos Bovinos , Feminino , Proteína Antagonista do Receptor de Interleucina 1/sangue , Proteína Antagonista do Receptor de Interleucina 1/metabolismo , Interleucina-1beta/metabolismo , Interleucina-4/sangue , Interleucina-4/metabolismo , Interleucina-8/sangue , Interleucina-8/metabolismo , Cistos Ovarianos/veterinária , Folículo Ovariano/metabolismo , Folículo Ovariano/patologia , Receptores Tipo I de Interleucina-1/metabolismo , Receptores Tipo II de Interleucina-1/metabolismo
15.
JBRA Assist Reprod ; 23(4): 367-391, 2019 10 14.
Artigo em Inglês | MEDLINE | ID: mdl-31173497

RESUMO

OBJECTIVE: The follicular fluid (FF) of women with polycystic ovary syndrome (PCOS) seems to exhibit a profile different from that of fertile women, which may be related to folliculogenesis disruption in PCOS patients. The aim of this study was to evaluate the differentially expressed proteins in the FF of women with PCOS compared to oocyte donors (ODs). METHODS: This screening study included thirteen (13) women who underwent in vitro fertilization (IVF) cycles: seven (7) ODs and six (6) PCOS patients. The patients underwent standard ovarian stimulation, and the FF was analysed using ion trap and time-of-flight liquid chromatography-mass spectrometry (LCMS-IT-TOF). RESULTS: The FF of the patients was matched to 229 proteins, with 61 proteins exclusive to the PCOS group, 123 proteins exclusive to the ODs, and 45 proteins found in both groups. We highlight fetuin-A and vitamin D ligand protein, which were exclusively expressed in the PCOS group; Complement C3 overexpressed in the PCOS group; and 26S protease only expressed in the OD group. The canonical pathways LXR/RXR activation, FXR/RXR activation, prothrombin activation are directly related to the disrupted metabolism and increased inflammatory status found in PCOS patients. CONCLUSIONS: The findings of the differentially expressed proteins and matched pathways are associated with folliculogenesis, indicating it relevance to oocyte quality.


Assuntos
Fertilização in vitro , Líquido Folicular/metabolismo , Oócitos/metabolismo , Síndrome do Ovário Policístico/metabolismo , Adulto , Feminino , Humanos , Doação de Oócitos , Indução da Ovulação , Estudos Prospectivos , Proteômica , Doadores de Tecidos , Adulto Jovem
16.
Mol Cell Endocrinol ; 492: 110444, 2019 07 15.
Artigo em Inglês | MEDLINE | ID: mdl-31075302

RESUMO

Studies in mice have shown that C-type natriuretic peptide (CNP) is produced by granulosa cells and contributes to ovarian follicle growth and oocyte meiotic arrest until the preovulatory LH surge. In humans, the relationship between intraovarian CNP levels and oocyte meiotic resumption is unknown. The aim of this study was to investigate whether CNP and its receptor NPR2 are expressed in human ovarian follicles and if their levels change according to the meiotic phase of oocytes. We collected follicular fluid (FF) and luteinized granulosa cells (LGC) from follicle pools (n = 47), and FF, LGC and cumulus cells (CC) from individual follicles (n = 96) during oocyte pickup for in vitro fertilization. There was a positive linear correlation between CNP levels in FF pools and basal antral follicle counting (rs = 0.458; p = 0.002), number of preovulatory follicles >16 mm (rs = 0.361; p = 0.016) and number of oocytes retrieved (rs = 0,378; p = 0.011) and a negative correlation between CNP levels in FF pools and the percentage of mature (MII) oocytes retrieved (rs = -0.39; p = 0.033). FF CNP levels in follicles containing MII oocytes were significantly lower than in follicles containing immature (MI) oocytes (median = 0.44 vs. 0.57 ng/mL, p < 0.05). Accordingly, the CNP precursor gene NPPC was 50% less expressed in LGC from follicles containing MII oocytes than in follicles containing MI oocytes (p < 0.01). In addition, NPR2 mRNA was down-regulated in CC surrounding MII oocytes (60% reduction, p < 0.01). CNP signaling is downregulated in human ovarian follicles containing mature oocytes. Further studies should clarify whether CNP signaling is essential to keep oocyte meiotic arrest in humans.


Assuntos
Regulação para Baixo , Peptídeo Natriurético Tipo C/genética , Peptídeo Natriurético Tipo C/metabolismo , Oócitos/fisiologia , Receptores do Fator Natriurético Atrial/genética , Receptores do Fator Natriurético Atrial/metabolismo , Adulto , Estudos Transversais , Células do Cúmulo/metabolismo , Feminino , Líquido Folicular/metabolismo , Células da Granulosa/metabolismo , Humanos , Meiose , Folículo Ovariano/metabolismo , Estudos Prospectivos , Transdução de Sinais
17.
J Assist Reprod Genet ; 36(5): 1003-1011, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31011990

RESUMO

PURPOSE: The present study aimed to provide a non-invasive approach to studying mechanisms responsible for oocyte development. METHODS: To this end, follicular fluid (FF) from 62 patients undergoing in vitro fertilization (IVF) cycles was split into two groups depending on the pregnancy outcome: pregnant (n = 28) and non-pregnant (n = 34) groups. Data were acquired by the MALDI-TOF mass spectrometry. Principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) were applied to the data set. A ROC curve, to predict success rate, was constructed, and the lipids were attributed. RESULTS: Six ions were differentially represented in FF of pregnant and non-pregnant patients, with an area under the curve of 0.962. Phosphatidic acid, phosphatidylglycerol, and triacylglycerol were hyper-represented in the pregnant group, while glucosylceramide was hyper-represented in the non-pregnant group. Enriched functions related to these lipids are steroidogenesis, cellular response, signal transduction, cell cycle, and activation of protein kinase C for the pregnant group and apoptosis inhibition for the non-pregnant group. CONCLUSION: Human FF fingerprinting can both improve the understanding concerning mechanisms responsible for oocyte development and its effect on embryo implantation potential and assist in the management of IVF cycles.


Assuntos
Biomarcadores/análise , Implantação do Embrião , Fertilização in vitro/métodos , Líquido Folicular/metabolismo , Lipídeos/análise , Oócitos/metabolismo , Resultado da Gravidez , Adulto , Feminino , Humanos , Oócitos/citologia , Oogênese , Indução da Ovulação , Valor Preditivo dos Testes , Gravidez
18.
JBRA Assist Reprod ; 23(2): 172-174, 2019 04 30.
Artigo em Inglês | MEDLINE | ID: mdl-30614664

RESUMO

We describe a case of a 37-year-old female, indicated for in vitro fertilisation. She developed skin rash on her trunk and limbs, during the treatment. RT-PCR results were positive in her blood and negative in her husband's blood and semen. Oocyte aspiration was performed, retrieving 7 oocytes, follicular fluid, and cumulus cells. RT-PCR results for the follicular fluid and cumulus cells were negative for ZIKV, and positive for only 2 oocytes. This is the first report in the literature analysing ZIKV in the follicular fluid, cumulus cells, and oocytes, and will contribute to the understanding of ZIKV infection and transmission.


Assuntos
Oócitos/virologia , Folículo Ovariano/virologia , Indução da Ovulação , Infecção por Zika virus , Zika virus/genética , Adulto , Feminino , Humanos , Masculino , RNA Viral/genética , Reação em Cadeia da Polimerase em Tempo Real , Sêmen/virologia , Infecção por Zika virus/diagnóstico , Infecção por Zika virus/transmissão , Infecção por Zika virus/virologia
19.
JBRA Assist Reprod, v. 23, n. 4, p. 367-391, oct. 2019
Artigo em Inglês | Sec. Est. Saúde SP, SESSP-IBPROD, Sec. Est. Saúde SP | ID: bud-2904

RESUMO

OBJECTIVE: The follicular fluid (FF) of women with polycystic ovary syndrome (PCOS) seems to exhibit a profile different from that of fertile women, which may be related to folliculogenesis disruption in PCOS patients. The aim of this study was to evaluate the differentially expressed proteins in the FF of women with PCOS compared to oocyte donors (ODs). METHODS: This screening study included thirteen (13) women who underwent in vitro fertilization (IVF) cycles: seven (7) ODs and six (6) PCOS patients. The patients underwent standard ovarian stimulation, and the FF was analysed using ion trap and time-of-flight liquid chromatography-mass spectrometry (LCMS-IT-TOF). RESULTS: The FF of the patients was matched to 229 proteins, with 61 proteins exclusive to the PCOS group, 123 proteins exclusive to the ODs, and 45 proteins found in both groups. We highlight fetuin-A and vitamin D ligand protein, which were exclusively expressed in the PCOS group; Complement C3 overexpressed in the PCOS group; and 26S protease only expressed in the OD group. The canonical pathways LXR/RXR activation, FXR/RXR activation, prothrombin activation are directly related to the disrupted metabolism and increased inflammatory status found in PCOS patients. CONCLUSIONS: The findings of the differentially expressed proteins and matched pathways are associated with folliculogenesis, indicating it relevance to oocyte quality.

20.
JBRA Assist Reprod ; 22(4): 346-351, 2018 11 01.
Artigo em Inglês | MEDLINE | ID: mdl-30264946

RESUMO

OBJECTIVE: To investigate the possible effect of follicular fluid glycodelin levels on the quality of developing oocytes and subsequent in vitro embryo development. METHODS: Follicular fluid glycodelin levels of 145 patients undergoing assisted reproductive treatment were analyzed and the correlation between glycodelin levels and ART outcomes were evaluated. RESULTS: We found that glycodelin levels were negatively correlated with the number of high quality embryos on day 3 (r=-0.20, p=0.05). Additionally, higher glycodelin levels were correlated with higher FSH levels (r=0.18, p=0.04). However, glycodelin levels were not predictive for implantation (p=0.67) or ongoing pregnancy rates (p=0.99). CONCLUSION: Glycodelin in the follicular environment might be one of the factors that influence the competence of growing oocytes and affect the quality of subsequent in vitro embryo development.


Assuntos
Desenvolvimento Embrionário , Líquido Folicular/metabolismo , Glicodelina/metabolismo , Adulto , Feminino , Hormônio Foliculoestimulante/sangue , Humanos , Gravidez , Resultado da Gravidez , Técnicas de Reprodução Assistida
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