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1.
Ceska Gynekol ; 85(1): 11-14, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32414279

RESUMO

OBJECTIVE: Description of punction of follicular fluid in a patient after ovarian transposition. DESIGN: Case report. SETTING: Department of Obstetrics and Gynaecology, 1st Faculty of Medicine, Charles University and the General Faculty Hospital, Prague. CASE REPORT: We present a case of IVF treatment in a patient with ovarian transposition undergoing punction of follicular fluid and difficulties during this procedure acording to transabdominal route. CONCLUSION: Transabdominal punction od follicular fluid is possible, but with technical difficulities and smaller amount of obtained oocytes. We recomend to aplicate IVF procedures prior to surgical solution.


Assuntos
Cavidade Abdominal/cirurgia , Fertilização In Vitro , Líquido Folicular/química , Procedimentos Cirúrgicos em Ginecologia/métodos , Recuperação de Oócitos/métodos , Folículo Ovariano/cirurgia , Feminino , Humanos , Oócitos , Gravidez
2.
Sheng Wu Gong Cheng Xue Bao ; 36(4): 632-642, 2020 Apr 25.
Artigo em Chinês | MEDLINE | ID: mdl-32347058

RESUMO

Extracellular vesicles (EVs) refer to bilayer membrane transport vesicles secreted by cells. EVs can take macromolecules from cells and transfer them to receptor cells. Among these macromolecular substances, the most studied are microRNAs (miRNAs). miRNA is non-coding RNA involved in the regulation of gene expression. It has been confirmed that there are different non-coding RNAs in mammalian follicular fluid EVs. EVs carrying miRNA can act as an alternative mechanism for autocrine and paracrine, affecting follicular development. This paper systematically introduced the kinds, characteristics and methods of isolation and identification of EVs, focusing on the effects of EVs and miRNAs on follicular development, including early follicular development, oocyte maturation, follicular dominance and effects on granulosa cell function. At the same time, the authors prospected the future research of EVs and microRNAs in follicular fluid, and provided ideas and directions for the research and application of EVs and miRNA functions in follicular fluid.


Assuntos
Vesículas Extracelulares , Líquido Folicular , MicroRNAs , Oogênese , Animais , Vesículas Extracelulares/metabolismo , Feminino , Líquido Folicular/química , Células da Granulosa/efeitos dos fármacos , MicroRNAs/farmacologia , Oogênese/efeitos dos fármacos
3.
Theriogenology ; 141: 120-127, 2020 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-31536861

RESUMO

Mastitis has deleterious effects on ovarian function and reproductive performance. We studied the association between plasma or follicular fluid (FF) obtained from endotoxin-induced mastitic cows, and oocyte developmental competence. Lactating Holstein cows were synchronized using the Ovsynch protocol. On Day 6 of the synchronized cycle, an additional PGF2α dose was administered, and either Escherichia coli endotoxin (LPS, 10 µg; n = 3 cows) or saline (n = 3 cows) was administered to one udder quarter per cow, 36 h later. Milk samples were collected and rectal temperatures recorded. Cows treated with LPS showed a typical transient increase in body temperature (40.3 °C ±â€¯0.4), whereas cows treated with saline maintained normal body temperature (38.9 °C ±â€¯0.04). A higher (P < 0.05) somatic cell count was recorded for cows treated with LPS. Plasma samples were collected and FF was aspirated from the preovulatory follicles by transvaginal ultrasound probe, 6 h after LPS administration. Radioimmunoassay was performed on plasma samples to determine estradiol and cortisol concentrations. Either FF or plasma was further used as maturation medium. In the first experiment, oocytes were matured in TCM-199 (Control) or in FF aspirated from cows treated with saline (FF-Saline) or LPS (FF-LPS). Cleavage rate to the 2- to 4-cell stage embryo did not differ among groups. However, the proportion of developed blastocysts on Day 7 postfertilization in the FF-LPS group tended to be lower for that in FF-Saline and was lower (P < 0.05) than that in the Control groups (10.6 vs. 22.4 and 24.4%, respectively). In the second experiment, oocytes were matured in TCM-199 (Control), or in plasma obtained from cows treated with saline (Plasma-Saline) or LPS (Plasma-LPS). Similar to the FF findings, cleavage rate did not differ among groups; however, the proportion of developing blastocysts tended to be lower in the Plasma-LPS group than in the Plasma-Saline group and was lower (P < 0.05) from that in the Control group (11.0 vs. 25.5 and 34.7%, respectively). The proportion of apoptotic cells per blastocyst, determined by TUNEL assay, did not differ among the experimental groups. The findings shed light on the mechanism by which mastitis induces a disruption in oocyte developmental competence. Further studies are required to clarify whether the negative effect on oocyte developmental competence is a result of LPS, by itself, or due to elevation of secondary inflammatory agents.


Assuntos
Bovinos , Líquido Folicular/química , Lipopolissacarídeos/toxicidade , Mastite Bovina/induzido quimicamente , Oócitos/efeitos dos fármacos , Plasma , Animais , Meios de Cultura , Fragmentação do DNA , Feminino , Técnicas de Maturação in Vitro de Oócitos/veterinária
4.
Anim Reprod Sci ; 211: 106208, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31785624

RESUMO

This study was conducted to investigate effects of short-term nutritional supplementation on concentrations of reproductive hormones in follicular fluid and on mRNA abundance in granulosa cells (GCs) during the luteal phase of ewes. Eighteen ewes were randomly assigned to treatment or control groups (n = 9, each group). All the ewes were subjected to estrous synchronization using a Controlled Intravaginal Releasing Device (CIDR). From the second to the eleventh day of estrous synchronization, ewes were fed a diet with a relatively greater (treatment group) or a maintenance (control group) energy content. Samples of follicular fluid and GCs were collected from antral follicles of estrous ewes after CIDR removal. The results indicate mean FSH concentration of fluid in small follicles and mean LH concentrations of fluid in large follicles of the ewes in the treatment group were greater (P < 0.05) than that of ewes in the control group. Follicular fluid E2 concentrations were directly related (P < 0.05) to follicular diameter. The ewes of the treatment group had greater (P < 0.05), compared with the control group, abundances of Follicle Stimulating Hormone Receptor (FSHR) in small and medium follicles, and (P<0.05) Luteinizing Hormone Receptor (LHR), Steroid Acute Regulatory Protein (STAR), cytochrome P450 (CYP17A1, CYP19A1) enzyme and Estrogen Receptor (ESR1) in large follicles. The results of this study provide evidence for a potential mechanism by which short-term nutritional supplementation improves follicular development possibly because of increased expression of steroid synthesis-regulating genes in large follicles.


Assuntos
Suplementos Nutricionais , Ingestão de Energia , Líquido Folicular/química , Células da Granulosa/metabolismo , RNA Mensageiro/metabolismo , Ovinos/fisiologia , Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal , Animais , Dieta/veterinária , Sincronização do Estro , Feminino , Hormônio Foliculoestimulante/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Folículo Ovariano/metabolismo
5.
J Pak Med Assoc ; 69(Suppl 3)(8): S4-S8, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31603867

RESUMO

Objectives: To evaluate the effect of acupuncture at follicular phase of menstrual cycle on anti-mullerian hormone levels in patients with polycystic ovary syndrome undergoing in-vitro fertilisation and to see its impact on assisted reproduction outcome. Methods: The prospective, randomised, controlled trial was conducted from March 2011 to July 2012 at the Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China. In the center, the patients randomly chose odd or pair number, the patients with odd numbers classified as an interventional group and the patients with paired numbers as non-interventional group. Infertile polycystic ovary syndrome patients aged 20-40 years were enrolled from the hospital's Assisted Reproduction Centre from March 2011 to July 2012. The patients were randomised into two groups, with one receiving follicular phase acupuncture for 30-40 minutes according to the principles of traditional Chinese medicine, and the other group not getting subjected to acupuncture. Serum and follicular anti-mullerian hormone concentration were determined. Results: Of the 102 patients, 33(32.4%) were in the intervention group, while 69(67.6%) were in the control group. There was no significant effect of acupuncture on serum and follicular fluid anti-mullerian hormone levels in the intervention group compared to the control group (p>0.05). Serum progesterone and estradiol levels on the day of giving human chorionic gonadotrophin, as well as serum progesterone and estradiol levels on the day of oocytes pick-up were significantly lower in the intervention group (p<0.05). Number of embryos transferred, clinical and ongoing pregnancy rates were significantly higher in the intervention group (p<0.05) with a significant decrease of ovarian hyper-stimulation syndrome rate in the intervention group (p<0.05). Conclusions: Follicular phase acupuncture was found to have a positive effect for polycystic ovary syndrome patients undergoing in-vitro fertilisation, but it had no effect on anti mullerian hormone concentrations.


Assuntos
Terapia por Acupuntura , Hormônio Antimülleriano/análise , Fertilização In Vitro , Infertilidade Feminina/terapia , Síndrome de Hiperestimulação Ovariana/prevenção & controle , Síndrome do Ovário Policístico/sangue , Hormônio Antimülleriano/sangue , Gonadotropina Coriônica/efeitos adversos , Gonadotropina Coriônica/uso terapêutico , Estradiol/sangue , Feminino , Líquido Folicular/química , Humanos , Infertilidade Feminina/etiologia , Síndrome de Hiperestimulação Ovariana/etiologia , Síndrome do Ovário Policístico/complicações , Gravidez , Taxa de Gravidez , Progesterona/sangue , Estudos Prospectivos
6.
Anim Reprod Sci ; 209: 106142, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31514932

RESUMO

Haemorrhagic anovulatory follicles (HAFs) are the most common pathological anovulatory condition in the mare. To enhance understanding of the physiopathology of HAFs, the aim of the present study was to determine the effects of an induced-follicular wave on LH concentrations and follicular fluid factors relevant to the ovulatory process. Mares were allocated to treatment or control groups (n = 7/group) in a crossed over design during 14 oestrous cycles with a period of one cycle occurring when there were no treatments between the times when treatments were administered. In the treatment group, all antral follicles ≥8 mm were ablated on Day 10 after ovulation followed by administration of a luteolytic dose of PGF2α. All mares of both groups were treated with 1500 IU of hCG when a follicle ≥32 mm was detected (Hour 0), and follicular fluid was aspirated 35 h later. Blood samples were collected every 48 h from Day 10 until Hour 0 from all mares. Follicular fluid was assayed for PGE2, estradiol and progesterone. Plasma was assayed for LH concentrations. A follicular wave followed follicle ablation in the treated mares. Concentrations of LH were greater (P = 0.05) in mares ot the treatment compared with control group. Concentrations of PGE2, estradiol and progesterone in follicular fluid did not differ between groups (P > 0.05). Treatment resulted in an earlier increase in circulating LH, however, there was no effect on concentrations of intra-follicular PGE2, estradiol or progesterone in hCG-stimulated preovulatory follicles.


Assuntos
Técnicas de Ablação , Anovulação/cirurgia , Líquido Folicular/metabolismo , Cavalos , Hormônio Luteinizante/sangue , Luteólise/efeitos dos fármacos , Folículo Ovariano/cirurgia , Técnicas de Ablação/métodos , Técnicas de Ablação/veterinária , Animais , Anovulação/complicações , Anovulação/metabolismo , Anovulação/veterinária , Gonadotropina Coriônica/farmacologia , Estudos Cross-Over , Dinoprosta/farmacologia , Ciclo Estral/efeitos dos fármacos , Ciclo Estral/metabolismo , Feminino , Líquido Folicular/química , Líquido Folicular/efeitos dos fármacos , Hemorragia/complicações , Hemorragia/cirurgia , Hemorragia/veterinária , Doenças dos Cavalos/metabolismo , Doenças dos Cavalos/cirurgia , Folículo Ovariano/diagnóstico por imagem , Folículo Ovariano/patologia , Ovulação/efeitos dos fármacos , Indução da Ovulação/métodos , Indução da Ovulação/veterinária , Punções/métodos , Punções/veterinária , Ultrassonografia de Intervenção/métodos , Ultrassonografia de Intervenção/veterinária
7.
J Vet Med Sci ; 81(9): 1313-1317, 2019 Sep 18.
Artigo em Inglês | MEDLINE | ID: mdl-31341113

RESUMO

We investigated the association among endometrial hyperemia, uterine bacterial infection, and features of the large ovarian follicles in dairy cows. Genital organs were collected in a complete set at a slaughterhouse, and the degree of endometrial hyperemia was examined for the direct evaluation of uterine inflammation. The rate of bacterial infection in the uterus was higher in cows with endometrial hyperemia regardless of the severity of hyperemia, compared with cows without hyperemia. Moreover, the characteristics of the follicular fluid were changed in cows with uterine bacterial infection and included high concentrations of lipopolysaccharide and malondialdehyde (lipid peroxidation marker). These findings can be utilized as the basic information for the direct evaluation of the uterine inflammatory status in dairy cows.


Assuntos
Doenças dos Bovinos/microbiologia , Folículo Ovariano/fisiopatologia , Doenças Uterinas/veterinária , Animais , Bactérias/isolamento & purificação , Bovinos , Endometrite/veterinária , Feminino , Líquido Folicular/química , Hiperemia/veterinária , Inflamação , Lipopolissacarídeos/análise , Malondialdeído/análise , Doenças Uterinas/microbiologia
8.
Theriogenology ; 136: 111-117, 2019 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-31254724

RESUMO

We aimed to characterize the expression of angiogenesis-related genes in corpus luteum (CL) and cumulus cells (CC) during estrous cycle in gilts of different genetic groups, as well as to study the relation between gene expression and phenotypic data. Forty five gilts were used as follows: L1, Commercial Line 1 (Large White x Landrace x Duroc) (n = 15); L2, Commercial Line 2 (Large White x Landrace x Pietrain) (n = 15); and Piau, Piau breed gilts (n = 15). Estrus observation started from 120 days of age. After the second observed estrus females were slaughtered (n = 3) on days 3, 5, 10, 14 and 18 of estrous cycle (first day of estrous cycle as Day 0). CL sampling was performed on days 3, 5, 10 and 14 and collection of CC and follicular fluid on days 14 and 18. Follicular fluid was used for analysis of estradiol levels and CC and CL samples for analysis of angiogenesis-related genes expression, ANGPT-1/-2 and TEK in CC and MMP-2, VEGFA, VEGFR-1/-2, ANGPT-1/-2 and TEK in CL. Piau gilts showed lower ovulation rate than both L1 and L2 gilts (P < 0.05), lower number of large antral follicles (>6 mm) at 18 days than L2 gilts (P < 0.05), and smaller diameter of the largest follicles at 14 days than L1 gilts (P < 0.05). Piau and L2 gilts showed higher estradiol levels in follicular fluid on day 18. Expression of ANGPT-1 and -2 genes in CC did not differ among genetic groups neither among days of the estrous cycle, but TEK gene expression was higher in L1 than L2 gilts on day 18. Expression of VEGFA, VEGFR-2 and MMP-2 genes in CL did not differ among genetic groups and days of cycle, but VEGFR-1 expression was higher in Piau than L2 gilts on days 10 and 14, and it was higher in L1 than L2 gilts on day 14. The ANGPT-1/-2 and TEK genes expression in CL were significantly higher in Piau than L1 gilts on day 10. The ANGPT-2/ANGPT-1 gene expression ratio in CL was higher in L1 than Piau and L2 gilts at 14 days, suggesting a shorter luteal phase for L1 gilts. Results indicated differences among genetic groups for the pattern of the angiogenesis-related genes expression in CL along estrous cycles, which may be reflected in phenotypic traits such as ovulation rate, estradiol levels in follicular fluid and number and diameter of antral follicles.


Assuntos
Corpo Lúteo/metabolismo , Células do Cúmulo/metabolismo , Regulação da Expressão Gênica/fisiologia , Suínos/genética , Suínos/fisiologia , Animais , Cruzamento , Estradiol/química , Estradiol/metabolismo , Feminino , Líquido Folicular/química
9.
PLoS One ; 14(6): e0217760, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31188849

RESUMO

Extracellular vesicles such as exosomes contain several types of transcripts, including mRNAs and micro RNAs (miRNAs), and have emerged as important mediators of cell-to-cell communication. Exosome-like vesicles were identified in the ovarian follicles of several mammalian species. Although the miRNA contents have been extensively characterized, the detailed investigation of their mRNA profiles is lacking. Here, we characterize the mRNA profiles of exosome-like vesicles in ovarian follicles in a pig model. The mRNA contents of the exosome-like vesicles isolated from porcine follicular fluid were analyzed and compared with those from mural granulosa cells (MGCs) using the Illumina HiSeq platform. Bioinformatics studies suggested that the exosomal mRNAs are enriched in those encoding proteins involved in metabolic, phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K) -protein kinase B (AKT), and mitogen-activated protein kinase (MAPK) pathways. While the mRNA profile of the exosome-like vesicles resembled that of MGCs, the vesicles contained mRNAs barely detectable in MGCs. Thus, while the majority of the vesicles are likely to be secreted from MGCs, some may originate from other cell types, including theca cells and oocytes, as well as the cells of non-ovarian organs/tissues. Therefore, the mRNA profiles unveiled several novel characteristics of the exosome-like vesicles in ovarian follicles.


Assuntos
Exossomos/genética , Líquido Folicular/metabolismo , Células da Granulosa/metabolismo , RNA Mensageiro/genética , Células Tecais/metabolismo , Transcriptoma , Animais , Comunicação Celular , Biologia Computacional/métodos , Exossomos/química , Exossomos/metabolismo , Feminino , Líquido Folicular/química , Expressão Gênica , Perfilação da Expressão Gênica , Células da Granulosa/química , Redes e Vias Metabólicas , Proteínas Quinases Ativadas por Mitógeno/genética , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Oócitos/química , Oócitos/metabolismo , Especificidade de Órgãos , Fosfatidilinositol 3-Quinases/genética , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/genética , Proteínas Proto-Oncogênicas c-akt/metabolismo , RNA Mensageiro/classificação , RNA Mensageiro/metabolismo , Suínos , Células Tecais/química
10.
J Biol Regul Homeost Agents ; 33(3): 946-956, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31190512

RESUMO

Female fertility plays a decisive role in the reproduction of mammals, with related issues that include oocyte or embryo quality, establishment of pregnancy, and the physiology of the tissues that contribute to reproduction and metabolic disorders associated with reproductive failure. Although reproductive failure may be attributed to various factors in different species, female infertility is largely controlled by a number of molecular signals that can be regulated in a cycle- and tissue-dependent manner.


Assuntos
Líquido Folicular/química , MicroRNAs/análise , Técnicas de Reprodução Assistida , Feminino , Humanos , Infertilidade Feminina/terapia , Oócitos , Gravidez
11.
Reprod Biol Endocrinol ; 17(1): 47, 2019 Jun 19.
Artigo em Inglês | MEDLINE | ID: mdl-31217014

RESUMO

BACKGROUND: Human follicular fluid is an intricate biological fluid contributing to the developing oocyte microenvironment. Accumulating evidence suggests that sex hormones present in follicular fluid (FF) may play an important role in regulating oocyte developmental potential. The aim of this study was to determine if anti-Müllerian hormone (AMH) and progesterone (P4) levels in FF are correlated with oocyte quality as defined by subsequent embryonic development. METHODS: This was a prospective cohort study of 88 women undergoing IVF/ICSI at a university associated fertility clinic. Follicular fluid was collected from the first follicle aspirated at the time of oocyte retrieval. The corresponding oocyte was individually cultured in order to track its developmental outcome. FF-AMH and P4 concentrations from follicles where the oocyte fertilised normally and developed into a blastocyst on day 5 (Group 1: BLAST, n = 23) were compared with FF from follicles where the oocyte fertilised normally but failed to reach blastocyst stage by day 5 (Group 2: FERT, n = 19). No significant differences were observed between the two groups in terms of maternal age, body mass index, previous live births, previous pregnancy loss, number of antral follicles, number of oocytes recovered, IVF:ICSI ratio or percentage of recovered oocytes that fertilised. RESULTS: FF-AMH and P4 levels were significantly increased in Group 1: BLAST compared to Group 2: FERT (P = 0.007 and P = 0.013 respectively). Twenty-one FF samples had an AMH level > 15 pmol/L, of which 17 related to oocytes that progressed to blastocyst stage, providing a positive prediction value (PPV) of 76.96%. Eleven FF samples had a P4 level > 60 mg/ml, of which 10 progressed to blastocyst stage, providing a PPV of 90.99%. Six samples had an AMH level > 15 pmol/L and a P4 level > 60 mg/ml, of which 100% progressed to blastocyst stage, providing a PPV of 96.83%. CONCLUSIONS: FF-AMH and P4 levels from individual follicles can accurately predetermine subsequent embryonic development. Combining follicular fluid analysis with routine morphological assessment, could allow for a more accurate and sensitive method of determining embryonic developmental competence.


Assuntos
Hormônio Antimülleriano/metabolismo , Desenvolvimento Embrionário , Fertilização In Vitro/métodos , Líquido Folicular/química , Progesterona/metabolismo , Adulto , Blastocisto/citologia , Blastocisto/metabolismo , Feminino , Humanos , Recuperação de Oócitos , Oócitos/citologia , Oócitos/metabolismo , Folículo Ovariano/citologia , Folículo Ovariano/metabolismo , Valor Preditivo dos Testes , Gravidez , Taxa de Gravidez , Estudos Prospectivos
12.
Endocrinology ; 160(10): 2298-2313, 2019 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-31211369

RESUMO

Oocyte-secreted factors bone morphogenetic protein 15 (BMP15) and growth differentiation factor 9 (GDF9) are critical for folliculogenesis and fertility. This study developed ELISAs for the measurement of BMP15 and GDF9 in serum and investigated their usefulness as biomarkers of female reproductive function. Serum samples were obtained from women undergoing infertility treatments (n = 154) and from perimenopausal and postmenopausal women (n = 28). Serum concentrations of BMP15 and GDF9 were analyzed in women relative to age, anti-Müllerian hormone, number of oocytes retrieved, and polycystic ovary syndrome (PCOS) after superovulation for in vitro fertilization. BMP15 and GDF9 immunoassays were validated for specificity, sensitivity (24 and 26 pg/mL, respectively), and reproducibility. BMP15 and GDF9 were detectable in 61% and 29% of women, respectively. BMP15 and GDF9 varied 64-fold and 15-fold, respectively, between women, but they did not change within subjects following ovarian stimulation with gonadotropins. Serum GDF9 concentration, but not BMP15 concentration, was associated with oocyte number retrieved in patients without PCOS (P = 0.018). GDF9 and BMP15 associations with oocyte number differed significantly (P < 0.05) with PCOS status. GDF9 concentrations were lower in poor responders (women with fewer than four oocytes retrieved or with cancelled cycles; P = 0.020). Serum BMP15, but not GDF9, was lower in women >55 years of age, compared with women of reproductive age (P < 0.01). This study develops and validates immunoassays to quantitate BMP15 and GDF9 in human serum and to correlate concentrations with female reproductive potential. Although assay sensitivities require improvement, this study demonstrates the diagnostic potential of oocyte-secreted BMP15 and GDF9 as serum biomarkers in reproductive medicine.


Assuntos
Proteína Morfogenética Óssea 15/metabolismo , Fertilização In Vitro , Gonadotropinas/farmacologia , Fator 9 de Diferenciação de Crescimento/metabolismo , Infertilidade Feminina/metabolismo , Síndrome do Ovário Policístico/metabolismo , Adulto , Biomarcadores/sangue , Biomarcadores/química , Proteína Morfogenética Óssea 15/química , Proteína Morfogenética Óssea 15/genética , Ensaio de Imunoadsorção Enzimática , Feminino , Líquido Folicular/química , Regulação da Expressão Gênica/efeitos dos fármacos , Fator 9 de Diferenciação de Crescimento/química , Fator 9 de Diferenciação de Crescimento/genética , Humanos , Oócitos/metabolismo , Folículo Ovariano , Ovário/patologia , Síndrome do Ovário Policístico/sangue , Reprodutibilidade dos Testes , Superovulação
13.
Fertil Steril ; 112(2): 387-396.e3, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31146888

RESUMO

OBJECTIVE: To study whether increased body mass index is associated with altered expression of extracellular vesicle microRNAs (EV-linked miRNAs) in human follicular fluid. DESIGN: Cross-sectional study. SETTING: Tertiary-care university-affiliated center. PATIENT(S): One hundred thirty-three women undergoing in vitro fertilization (IVF) were recruited from January 2014 to August 2016. INTERVENTIONS(S): None. MAIN OUTCOME MEASURE(S): EV-linked miRNAs were isolated from follicular fluid and their expression profiles were measured with the use of the Taqman Open Array Human miRNA panel. EV-linked miRNAs were globally normalized and inverse-normal transformed. Associations between body mass index (BMI) and EV-linked miRNA outcomes were analyzed by means of multivariate linear regression and principal component analysis. RESULT(S): Eighteen EV-linked miRNAs were associated with an increase in BMI after adjusting for age, ethnicity, smoking status, and batch effects. Hsa-miR-328 remained significant after false discovery rate adjustments. Principal component analyses identified the first principal component to account for 40% of the variation in our EV-linked miRNA dataset, and adjusted linear regression found that the first principal component was significantly associated with BMI after multiple testing adjustments. Using Kyoto Encyclopedia of Genes and Genomes enrichment analyses, we predicted gene targets of EV-linked miRNA in silico and identified PI3K-Akt signaling, ECM-receptor interaction, focal adhesion, FoxO signaling, and oocyte meiosis pathways. CONCLUSION(S): These results show that a 1-unit increase in BMI is associated with altered follicular fluid expression of EV-linked miRNAs that may influence follicular and oocyte developmental pathways. Our findings provide potential insight into a mechanistic explanation for the reduced fertility rates associated with increased BMI.


Assuntos
Índice de Massa Corporal , Vesículas Extracelulares/metabolismo , Líquido Folicular/metabolismo , MicroRNAs/metabolismo , Adulto , Estudos Transversais , Vesículas Extracelulares/genética , Feminino , Fertilização In Vitro , Líquido Folicular/química , Humanos , MicroRNAs/análise , MicroRNAs/genética , Oogênese/genética , Transcriptoma , Adulto Jovem
14.
Trop Anim Health Prod ; 51(8): 2447-2454, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31197724

RESUMO

The objective of present study was to determine metabolite profile and inorganic elements of camel follicular fluids (FF) using "gas chromatography-mass spectrometry (GC-MS) and inductively coupled plasma mass spectrometry (ICP-MS)," respectively. Various metabolites were detected in camel FF by the proposed GC-MS technique. The major compounds detected were lactic acid (62.37%), linolenic acid (5.95%), myo-inositol (3.37%), hexadecanoic acid (3.19%), N-ethyl-N-vinylacetamide (3.15%), acetamide (2.89%), tetradecanoic acid (2.64%), and D-xylofuranose (2.25%). The proposed ICP-MS technique was validated in terms of linearity, precision, accuracy, and sensitivity. All quality control validation parameters were found to be satisfactory for the analysis of elements in camel FF. The proposed ICP-MS technique showed the presence of sixteen different elements (out of eighteen standards) in camel FF. Some elements such as Na, K, Ca, and Mg were obtained in higher amounts in camel FF. Overall, the results of this study indicated that the proposed GC-MS and ICP-MS techniques can be successfully applied for metabolite profile and element determination of biological fluids such as FF.


Assuntos
Camelus/metabolismo , Líquido Folicular/metabolismo , Animais , Feminino , Líquido Folicular/química , Cromatografia Gasosa-Espectrometria de Massas
15.
Reprod Biol Endocrinol ; 17(1): 45, 2019 Jun 11.
Artigo em Inglês | MEDLINE | ID: mdl-31186025

RESUMO

BACKGROUND: Polycystic ovary syndrome (PCOS) is a complex disorder associated with multiple metabolic disturbance, including defective glucose metabolism and insulin resistance. The altered metabolites caused by the related metabolic disturbance may affect ovarian follicles, which can be reflected in follicular fluid composition. The aim of this study is to investigate follicular fluid metabolic profiles in women with PCOS using an advanced sequential window acquisition of all theoretical fragment-ion spectra (SWATH) mass spectrometry. MATERIALS AND METHODS: Nineteen women with PCOS and twenty-one healthy controls undergoing IVF/ET were recruited, and their follicular fluid samples were collected for metabolomic study. Follicular fluid metabolic profiles, including steroid hormones, free fatty acids, bioactive lipids, and amino acids were analyzed using the principal component analysis (PCA) and partial least squares to latent structure-discriminant analysis (PLS-DA) model. RESULTS: Levels of free fatty acids, 3-hydroxynonanoyl carnitine and eicosapentaenoic acid were significantly increased (P < 0.05), whereas those of bioactive lipids, lysophosphatidylcholines (LysoPC) (16:0), phytosphingosine, LysoPC (14:0) and LysoPC (18:0) were significantly decreased in women with PCOS (P < 0.05). Additionally, levels of steroid hormone deoxycorticosterone and two amino acids, phenylalanine and leucine were higher in the PCOS patients (P < 0.05). CONCLUSION: Women with PCOS display unique metabolic profiles in their follicular fluid, and this data may provide us with important biochemical information and metabolic signatures that enable a better understanding of the pathogenesis of PCOS.


Assuntos
Biomarcadores/análise , Líquido Folicular/química , Espectrometria de Massas/métodos , Metabolômica/métodos , Folículo Ovariano/química , Síndrome do Ovário Policístico/diagnóstico , Adulto , Aminoácidos/análise , Ácidos Graxos não Esterificados/análise , Feminino , Humanos , Análise dos Mínimos Quadrados , Lipídeos/análise , Síndrome do Ovário Policístico/metabolismo , Análise de Componente Principal
16.
Nutrients ; 11(5)2019 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-31083424

RESUMO

BACKGROUND: Anti-Müllerian hormone (AMH) is considered to be one of the most significant indicators of women's fertility. Many studies have shown that vitamin D may modify human reproductive functions; however, the results are conflicting. The composition of follicular fluid (FF) creates the biochemical environment of the oocyte and affects its quality, which later determines the embryo quality. In this study, we aimed to revise with advanced statistical techniques the relationship between AMH and vitamin D in FF. METHODS: The study was designed as a prospective single-center study in infertile patients with AMH ≥ 0.7 ng/mL who underwent controlled ovarian hyperstimulation for in vitro fertilization. AMH and vitamin D levels in FF were measured. Next, the standard and advanced statistical (including segmented regression) techniques were applied. RESULTS: We observed a negative linear correlation between levels of AMH in serum and FF and total vitamin D concentrations up to approximately 30 ng/ml; with a statistically significant relationship in FF. Beyond that concentration, the trend was positive but statistically insignificant. CONCLUSIONS: As an existing "change-point problem" was noticed, we suggest segmentation in the relationship between vitamin D and AMH during infertility treatment.


Assuntos
Hormônio Antimülleriano/sangue , Infertilidade Feminina/terapia , Vitamina D/sangue , Adulto , Hormônio Antimülleriano/química , Hormônio Antimülleriano/metabolismo , Estudos de Coortes , Feminino , Fertilização In Vitro , Líquido Folicular/química , Humanos , Gravidez , Taxa de Gravidez , Estudos Prospectivos , Estações do Ano
17.
Gynecol Endocrinol ; 35(9): 803-806, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-30982370

RESUMO

Irisin, a novel exercise-induced myokine, has been implicated in different aspects of human metabolism and could be connected to polycystic ovary syndrome (PCOS). This study aimed to investigate serum and follicular fluid (FF) irisin levels in PCOS and normal women undergoing controlled ovarian stimulation and correlate them to the lipid and lipoprotein levels as well as with other metabolic parameters. Serum and FF irisin, together with serum lipid and lipoprotein levels were assessed in 70 women with diagnosed PCOS and 70 non-PCOS controls, under in vitro fertilization (IVF) treatment. Regardless of BMI, PCOS women had a significantly increased number of oocytes retrieved, fertilized oocytes and transferred embryos, although the number of women achieving pregnancies did not differ between groups. No correlation between FF irisin levels and pregnancy could be established. Serum and FF irisin levels were significantly higher in PCOS and overweight women and were positively associated with BMI and dyslipidemia. FF irisin levels correlated positively to and were lower than serum irisin levels. Further research would be helpful to analyze irisin's role in female reproduction, if any, as well as in human metabolism and the pathophysiology of PCOS.


Assuntos
Fibronectinas/sangue , Fibronectinas/metabolismo , Líquido Folicular/metabolismo , Lipoproteínas/sangue , Indução da Ovulação , Síndrome do Ovário Policístico/metabolismo , Adulto , Estudos de Casos e Controles , Dislipidemias/etiologia , Dislipidemias/metabolismo , Feminino , Líquido Folicular/química , Humanos , Infertilidade Feminina/etiologia , Infertilidade Feminina/metabolismo , Infertilidade Feminina/terapia , Resistência à Insulina/fisiologia , Metabolismo dos Lipídeos , Lipídeos/sangue , Indução da Ovulação/métodos , Síndrome do Ovário Policístico/sangue , Síndrome do Ovário Policístico/complicações , Gravidez
18.
Fertil Steril ; 111(5): 953-961, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30879714

RESUMO

OBJECTIVE: To explore whether follicular fluid (FF) concentrations of phthalate metabolites are associated with levels of intrafollicular reproductive hormones in women undergoing in vitro fertilization. DESIGN: Cross-sectional study. SETTING: University-affiliated reproductive medicine center. PATIENT(S): A total of 194 women each contributed one FF sample at oocyte retrieval. INTERVENTION(S): FF aspirates from individual follicles (≥18 mm in diameter) were collected. We measured eight phthalate metabolites and four ovarian hormones in the FF samples. MAIN OUTCOME MEASURE(S): Per-follicle E2, P, total T, and antimüllerian hormone (AMH) concentrations. RESULT(S): Most phthalate metabolites were highly detected in FF samples. We observed a dose-response relationship between increasing monomethyl phthalate (MMP) tertiles and lower E2, P, and T levels. Women in the third tertile of MMP had decreases of 34.23%, 9.44 ng/L, and 23.28% in E2, P, and T, respectively, compared with women in the first tertile. Tertiles of monoethyl phthalate and the percentage of di(2-ethylhexyl) phthalate) metabolites excreted as mono(2-ethylhexyl) phthalate were inversely associated with P. We also identified a negative relationship between monobenzyl phthalate tertiles and AMH. In addition, positive correlations between some of the phthalate metabolites and ovarian hormones were detected. CONCLUSION(S): FF concentrations of certain phthalate metabolites were associated with altered levels of intrafollicular reproductive hormones, which raises concern over a potential deleterious effect of environmental phthalate exposure on the endocrine capacity and viability of theca and granulosa cells.


Assuntos
Exposição Ambiental/efeitos adversos , Fertilização In Vitro/métodos , Líquido Folicular/metabolismo , Recuperação de Oócitos/métodos , Ácidos Ftálicos/metabolismo , Adulto , Estudos Transversais , Feminino , Líquido Folicular/química , Humanos , Pessoa de Meia-Idade , Ácidos Ftálicos/efeitos adversos , Ácidos Ftálicos/análise , Adulto Jovem
19.
Fertil Steril ; 111(6): 1217-1225.e3, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30922638

RESUMO

OBJECTIVE: To characterize ovarian follicles of girls and young women with Turner syndrome (TS) who underwent ovarian tissue cryopreservation (OTC). DESIGN: Retrospective case-control study. SETTING: University hospital. PATIENT(S): Fifteen girls and young women with TS aged 5-22 years at OTC were included, together with 42 control girls and young women aged 1-25 years who underwent OTC because of cancer. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Follicle density (follicles/mm3), morphology, and health were assessed in ovarian cortex biopsies from TS patients and compared with controls. Hormone concentrations were measured in serum and follicle fluids. Immature cumulus oocyte complexes were obtained and matured in vitro. RESULT(S): Follicles were found in 60% of the biopsies (9 of 15) from TS ovaries. In 78% of the ovaries (7 of 9) with follicles, the follicle density was within the 95% confidence interval of the control group. There was a high rate of abnormal follicle morphology. Six follicle-specific proteins were expressed similarly in TS and control ovaries. However, apoptosis and zona pellucida protein expression were found to be abnormal in TS. Turner syndrome follicle fluid from small antral follicles had lower concentrations of estrogen and testosterone and higher concentrations of antimüllerian hormone than controls. Thirty-one cumulus oocyte complexes were collected from one patient and cultured for 48 hours in vitro, resulting in five metaphase II oocytes (maturation rate 16%, degeneration rate 19%). CONCLUSION(S): The benefits of OTC may be limited to a highly selected group of TS mosaic patients in whom a sizeable pool of normal follicles is present at OTC.


Assuntos
Criopreservação , Preservação da Fertilidade/métodos , Infertilidade Feminina/terapia , Folículo Ovariano/patologia , Ovário/patologia , Técnicas de Reprodução Assistida , Síndrome de Turner/patologia , Adolescente , Adulto , Biópsia , Criança , Pré-Escolar , Tomada de Decisão Clínica , Feminino , Fertilidade , Líquido Folicular/química , Predisposição Genética para Doença , Humanos , Técnicas de Maturação in Vitro de Oócitos , Lactente , Infertilidade Feminina/etiologia , Infertilidade Feminina/patologia , Infertilidade Feminina/fisiopatologia , Folículo Ovariano/química , Folículo Ovariano/transplante , Ovário/química , Ovário/transplante , Seleção de Pacientes , Fenótipo , Valor Preditivo dos Testes , Estudos Retrospectivos , Síndrome de Turner/complicações , Síndrome de Turner/genética , Adulto Jovem
20.
EBioMedicine ; 41: 597-609, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30852161

RESUMO

BACKGROUND: The fallopian tube fimbria is regarded as the main tissue of origin and incessant ovulation as the main risk factor of ovarian high-grade serous carcinoma. Previously, we discovered the tumorigenesis activity of human ovulatory follicular fluid (FF) upon injection to the mammary fat pad of Trp53-null mice. We also found a mutagenesis activity of FF-ROS and a apoptosis-rescuing activity of Hb from retrograde menstruation. However, neither of them can explain the tumorigenesis activities of FF. METHODS: From two cohorts of ovulatory FF retrieved from IVF patients, the main growth factor responsible for the transformation of human fimbrial epithelial cells was identified. Mechanism of activation, ways of signal transduction of the growth factor, as well as the cellular and genetic phenotypes of the malignant transformation was characterized. FINDINGS: In this study, we showed that insulin-like growth factor (IGF)-axis proteins, including IGFBP-bound IGF2 as well as the IGFBP-lytic enzyme PAPP-A, are abundantly present in FF. Upon engaging with glycosaminoglycans on the membrane of fimbrial epithelial cells, PAPP-A cleaves IGFBPs and releases IGF2 to bind with IGF-1R. Through the IGF-1R/AKT/mTOR and IGF-1R/AKT/NANOG pathways, FF-IGF leads to stemness and survival, and in the case of TP53/Rb or TP53/CCNE1 loss, to clonal expansion and malignant transformation of fimbrial epithelial cells. By depleting each IGF axis component from FF, we proved that IGF2, IGFBP2/6, and PAPP-A are all essential and confer the majority of the transformation and regeneration activities. INTERPRETATION: This study revealed that the FF-IGF axis functions to regenerate tissue damage after ovulation and promote the transformation of fimbrial epithelial cells that have been initiated by p53- and Rb-pathway disruptions. FUND: The study was supported by grants of the Ministry of Science and Technology, Taiwan (MOST 106-2314-B-303-001-MY2; MOST 105-2314-B-303-017-MY2; MOST 107-2314-B-303-013-MY3), and Buddhist Tzu Chi General Hospital, Taiwan (TCMMP104-04-01).


Assuntos
Líquido Folicular/metabolismo , Fator de Crescimento Insulin-Like II/metabolismo , Animais , Carcinogênese , Aberrações Cromossômicas , Variações do Número de Cópias de DNA , Células Epiteliais/citologia , Células Epiteliais/metabolismo , Células Epiteliais/transplante , Tubas Uterinas/citologia , Feminino , Líquido Folicular/química , Humanos , Proteínas de Ligação a Fator de Crescimento Semelhante a Insulina/metabolismo , Camundongos , Camundongos Endogâmicos NOD , Camundongos Knockout , Proteína Plasmática A Associada à Gravidez/metabolismo , Interferência de RNA , RNA Interferente Pequeno/metabolismo , Receptores de Somatomedina/antagonistas & inibidores , Receptores de Somatomedina/genética , Receptores de Somatomedina/metabolismo , Transdução de Sinais , Proteína Supressora de Tumor p53/deficiência , Proteína Supressora de Tumor p53/genética
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