Composition Of Follicular Fluid Research Articles

Does Endometriosis Impact the Composition of Follicular Fluid in IL6 and AMH? A Case-Control Study

The aim of this study was to compare follicular liquid levels of IL6 and AMH in women with and without endometriosis and to evaluate their potential impact on ICSI outcomes. It is a prospective case-control study conducted on 25 women with proven endometriosis and 50 patients diagnosed with other causes of infertility. All these patients were candidates for ICSI cycles. Their follicular fluid was collected at the time of oocyte retrieval and used to evaluate IL-6 and AMH titers by electro-chemiluminescent immunoassay (Cobas e411-Roche). The IL-6 levels in follicular fluid were higher in the endometriosis group than in the control group (152.3 vs. 19.9 pg/mL; p = 0.02). The median level for AMH was 2.2 ± 1.88 ng/mL with no statistical difference between the two groups (2.2 vs. 2.7 ng/mL, p = 0.41). No significant correlation between the follicular IL6 and AMH levels was observed. The oocyte quality seems to be preserved in patients with endometriosis with the adequate response to ovarian stimulation. High levels of follicular IL6 are in accordance with the inflammatory phenomenon of the disease; however, this increase has no impact on ICSI outcomes.

Effect of PFOA exposure on diminished ovarian reserve and its metabolism

Owing to its difficulty in degrading and ease of accumulation in the body, perfluorooctanoic acid (PFOA) has a detrimental effect on reproduction. This study aimed to examine the effect of PFOA concentration in follicular fluid during ovulation stimulation on embryo quality and the impact of PFOA exposure on the metabolic components of follicular fluid. This was a single-center prospective study that included 25 patients with diminished ovarian reserve (DOR), 25 with normal ovarian reserve (NOR), and 25 with polycystic ovary syndrome (PCOS). Follicular fluid samples were analyzed using ultra-high performance liquid chromatography-tandem mass spectrometry. We demonstrated that the PFOA levels of follicular fluid in the DOR group were higher than those in the NOR group and PCOS group (P < 0.05). PFOA concentration in the PCOS group was negatively correlated with high-quality embryos (P < 0.05). To gain more insight into the impact of PFOA on the metabolic composition of follicular fluid, we classified the DOR group based on the PFOA concentration, for which metabolomic analysis was performed. In the high-concentration PFOA group, there was an increase and a decrease in three and nine metabolites, respectively, compared to that in the low-concentration group. These results suggest that PFOA may alter the metabolic composition of follicular fluid, thus, affecting ovarian reserve function.

Biochemical alterations in the follicular fluid of bovine peri-ovulatory follicles and association with final oocyte maturation.

Follicular fluid (FF), a product of vascular transudate and granulosa and thecal cell secretions, is the milieu that has evolved to support oocyte growth and maturation which plays a central role in oocyte quality determination. Therefore, a suboptimal FF composition may be reflected in compromised oocyte progression through maturation, fertilization or embryo development. To date, the composition of bovine FF remains understudied. To address this, we comprehensively characterized the metabolomic constituency of bovine FF in the period during which the oocyte undergoes meiotic maturation. More specifically, FF from pre (-24 h) and peri (-2 h) -ovulatory follicles was profiled by high-throughput untargeted ultra-high-performance liquid chromatography tandem mass spectroscopy. A total of 634 metabolites were identified, comprising: lipids (37.1%), amino acids (30.0%), xenobiotics (11.5%), nucleotides (6.8%), carbohydrates (4.4%), cofactors and vitamins (4.4%), peptides (3.6%) and energy substrates (2.1%). The concentrations of 67 metabolites were significantly affected by stage of follicle development, 33.3% (n=21) were reduced (P≤0.05) by a mean of 9.0-fold, whereas 46 were elevated (P≤0.05) by a mean of 1.7-fold in peri vs. pre -ovulatory FF. The most pronounced individual metabolite concentration decreases were hypoxanthine (98.9-fold), xanthine (65.7-fold), 17β-oestradiol (12.4-fold), and inosine (4.6-fold). In contrast, the greatest increases were in retinal (4.9-fold), 1-methyl-5-imidazoleacetate (2.7-fold), and isovalerylcarnitine (2.7-fold). This global metabolomic analysis of bovine FF temporal dynamics provides new information for understanding the environment supporting oocyte maturation and facilitating ovulation, that has the potential for improving oocyte quality both in vivo and in vitro.

Дескрипторы компетенции ооцитов и исходы программ экстракорпорального оплодотворения

This review presents data on estradiol, progesterone, and leptin as important hormones in reproductive medicine. The effects of these hormones in peripheral blood and follicular fluid on the outcomes of assisted reproductive technology programs are considered. Data on experimental studies and prospects for their use in clinical practice are presented. Given the established fact that the composition of follicular fluid has an influence on the quality of oocytes, estradiol, progesterone, and leptin contained in follicular fluid can serve as descriptors of embryo competence. There are conflicting results in the literature regarding the effect of these hormones on the outcomes of in vitro fertilization programs. Conclusion. The role of intrafollicular estradiol, progesterone, and leptin on the outcomes of in vitro fertilization programs need to be further investigated. Key words: follicular fluid, estradiol, progesterone, leptin, in vitro fertilization, embryo competence

Microbiological composition of follicular fluid in patients undergoing IVF and its association with infertility.

In recent years, the incidence of female infertility has risen sharply, which is affected by many factors. It was recognized that female reproductive tract microbes play a role in the process of female conception. If the reproductive tract microbes could solve a certain proportion of infertility, it would certainly reduce the pain and economic burden of many patients. The objective of this study was to investigate the microbial community composition of follicular fluid in infertile patients and its potential impact on infertility. Follicular fluid from 49 primary infertility and 52 secondary infertility patients was collected by a negative pressure needle, and the microbiota was analyzed by 16S rDNA sequencing. It was found that Lactobacillus, especially L. crispatus, might have a positive effect on female pregnancy. Considering the presence or absence of male factors and different body mass indices, L. iners might inhibit female pregnancy. However, L. iners seemed to play a positive role in egg maturation, while Gardnerella and Cutibacterium acnes might have a negative effect on female pregnancy. This study suggested the potential role of Lactobacillus in follicular fluid in improving female infertility and provided a theoretical basis for the future microbiological treatment of female infertility.

Obesity-related alterations in protein expression in human follicular fluid from women undergoing in vitro fertilization

To compare the proteomic composition of follicular fluid from women with normal weight vs. women with obesity but without a history of polycystic ovary syndrome or known ovarian dysfunction undergoing invitro fertilization. Cross-sectional. Academic medical center. Eight women with normal weight and 8 women with obesity undergoing invitro fertilization and without a history of polycystic ovary syndrome, ovulatory dysfunction, diminished ovarian reserve, or known endometriosis were included in the analysis. Not applicable. Proteomic assessment using liquid chromatography-mass spectrometry analysis. The mean age of women with normal weight was similar to that of women with obesity (32.9 vs. 32.6 years, not significant). The mean body mass index of women with normal weight was 21.2 kg/m2 compared with a body mass index of 37.1 kg/m2 in women with obesity. A total of 1,174 proteins were identified with ≥2 peptides present. Twenty-five proteins were found to be significantly altered in the follicular fluid from women with obesity. Of these 25 proteins, 19 were up-regulated and 6 were down-regulated. Notably, C-reactive protein was 11-fold higher in the follicular fluid from women with obesity than in the follicular fluid from women with normal weight. Obesity is associated with dysregulation at the level of the follicle, including alterations in proteins related to inflammation and metabolism. These include proteins with emerging roles in energy homeostasis and follicular regulation.

Follicular free fatty acid metabolic signatures and their effects on oocyte competence in non-obese PCOS patients.

Polycystic ovary syndrome (PCOS) is a common cause of anovulatory infertility in women. This study identified changes in free fatty acids profiles in the follicular fluid that may lead to better diagnosis and management of infertility in PCOS women. Polycystic ovary syndrome (PCOS) is a heterogeneous disease characterized by various endocrine/metabolic disorders and impaired reproductive potential. Alterations in oocyte competence are considered potentially causative factors for infertility in PCOS women and analyzing the composition of follicular fluid in these patients may help to identify which changes have the potential to alter oocyte quality. In this study, free fatty acid metabolic signatures in follicular fluid were performed to identify changes that may impact oocyte competence in non-obese PCOS women. Sixty-four non-obese women (32 with PCOS and 32 age- and BMI-matched controls) undergoing in vitro fertilization were recruited. Embryo quality was morphologically assessed. Free fatty acid metabolic profiling in follicular fluid was performed using gas/liquid chromatography-mass spectrometry. Principal component analysis and orthogonal partial least squares-discriminant analysis models were further constructed. Nine free fatty acids and 24 eicosanoids were identified and several eicosanoids synthesized by the cyclooxygenase pathway were significantly elevated in PCOS patients compared to controls. The combination of PGE2, PGF2α, PGJ2, and TXB2 had an area under the curve of 0.867 (0.775-0.960) for PCOS discrimination. Furthermore, follicular fluid levels of PGE2 and PGJ2 were negatively correlated with high-quality embryo rate in PCOS patients (P < 0.05). Metabolomic analysis revealed that follicular fluid lipidomic profiles undergo changes in non-obese PCOS women, which suggests that identifying changes in important metabolic signatures may give us a better understanding of the pathogenesis of PCOS. Furthermore, elevated PGE2 and PGJ2 concentrations may contribute to impaired oocyte competence in non-obese PCOS patients.

P-188 The effect of follicular fluid extracellular vesicles on motility and hyperactivation of human spermatozoa and differential miRNA levels depend on the woman's age

Abstract Study question How does the woman’s reproductive age affect the ability of follicular fluid (FF) extracellular vesicles (EV) to change the motility and hyperactivation of spermatozoa and follicular fluid microRNA profiles? Summary answer The effect of FF EVs on sperm motility and hyperactivation decreases with a woman's age. MiR-134-5p, miR-21-5p expression levels increase in older age group. What is known already Aging reduces human fertility. FF is an important factor in attracting and activating spermatozoa in the oviduct during oocyte fertilization, enhancing the process of sperm capacitation and acrosomal reaction in various mammalian species, including humans. EVs from seminal plasma and oviduct fluid carry proteins and miRNAs playing a vital role in a multi-step process including sperm motility, capacitation, acrosomal reaction, further fertilization. Results of our previous experiments showed that FF EVs from young women significantly improve the indices of sperm motility and hyperactivation. Several studies have isolated some age associated differentially expressed miRNAs (hsa-miR-424, hsa-miR-21-5p, hsa-miR-134, hsa-miR-190b, hsa-miR-99b -3p). Study design, size, duration FF EVs were obtained by sequential centrifugation at different rotational speeds, frozen at -80 °C. The sperm fraction was isolated, washed by differential centrifugation in a density gradient, suspended in the saline to a concentration of 106/ml and incubated with EVs (1:2) at 37 °C in CO2-incubator for 1 h. Samples were centrifuged and fixed in 2.5% glutaraldehyde in 0.1 M buffer for TEM. Sperm motility were assessed using CASA. MicroRNAs isolation (miRNeasy Serum/Plasma Kit (Qiagen)). Participants/materials, setting, methods All study participants signed a voluntary informed consent for the use of biological samples for research purposes. Sperm samples were isolated from seminal fluid (n = 18) aged 28-36 years without severe pathozoospermia. FF EVs were obtained from young (n = 4) aged 27-31 (EVs+) and older patients (n = 4) aged 41-46 with several IVF attempts (EVs-). The methods used in this work include sperm and EVs isolation, incubation, transmission electron microscopy, RT-PCR, statistical data analysis, computer-assisted semen analysis (CASA). Main results and the role of chance Our results showed that sperm incubation with EVs+ led to a significant increase in the number of progressively motile spermatozoa (paired Student's t test, p &amp;lt; 0.001), indicators of general mobility (p = 0.05) and hyperactivation of spermatozoa after 1 hour. Sperm incubation with EVs- slightly changed these parameters compared to the control (p = 0.171). Results from TEM showed that EVs- bind to spermatozoa worse than EVs+ after incubation. EVs- bind predominantly to the lateral membrane of the spermatozoon, rather than to the acrosomal region, compare to EVs+, which may reflect changes in the functional composition of FF EVs in older patients group and target areas of interaction with spermatozoa. This hypothesis is also supported by slight improvements in progressive sperm motility after incubation. Age-related changes are reflected in energy, metabolic and other important biological processes, which leads to a change in the composition and functions of FF (decrease in progesterone, glucose, increase in lactate and pyruvate, increase in reactive oxygen species) and affect the functional role of FF EVs. MiR-134-5p (p = 0.008), miR-21-5p (p = 0.008) expression levels were statistically significantly higher in EVs- group compared with EV + group with a successful first attempt. The expression of other microRNAs analyzed did not differ between groups. Limitations, reasons for caution Small amount of data. This study does not characterize isolated FF EVs in any way (specific markers, EVs concentration, size). To discuss the possible mechanisms underlying the effects of FF EVs of different age groups on sperm characteristics, it is necessary to perform metabolomics analyses of FF EVs. Wider implications of the findings Further studies of the influence of FF EVs on sperm motility may help to improve of ART outcomes, associated with male and female infertility by improving sperm morphofunctional characteristics and increasing their fertilizing ability. The solutions to this problem could be the use of FF EVs from young fertile donors. Trial registration number not applicable

Fatty Acids and Metabolomic Composition of Follicular Fluid Collected from Environments Associated with Good and Poor Oocyte Competence in Goats.

In goats, embryo oocyte competence is affected by follicle size regardless the age of the females. In previous studies we have found differences in blastocyst development between oocytes coming of small (<3 mm) and large follicles (>3 mm) in prepubertal (1–2 months-old) goats. Oocyte competence and Follicular Fluid (FF) composition changes throughout follicle growth. The aim of this study was to analyze Fatty Acids (FAs) composition and metabolomic profiles of FF recovered from small and large follicles of prepubertal goats and follicles of adult goats. FAs were analyzed by chromatography and metabolites by 1H-Nuclear Magnetic Resonance (1H-NMR) Spectrometry. The results showed important differences between adult and prepubertal follicles: (a) the presence of α,β-glucose in adult and no detection in prepubertal; (b) lactate, -N-(CH3)3 groups and inositol were higher in prepubertal (c) the percentage of Linolenic Acid, Total Saturated Fatty Acids and n-3 PUFAs were higher in adults; and (d) the percentage of Linoleic Acid, total MUFAs, PUFAs, n-6 PUFAs and n-6 PUFAs: n-3 PUFAs ratio were higher in prepubertal goats. Not significant differences were found in follicle size of prepubertal goats, despite the differences in oocyte competence for in vitro embryo production.

The Influence of the Corpus Luteum Location on Hormonal and Vitamin C Composition of Follicular Fluid and Serum in Dromedary Camels (Camelus dromedarius)

The aim of this study was to compare hormonal composition of the predominant antral follicle coexisting with or without a corpus luteum (CL) in dromedary camels. Forty-seven genitalia and blood samples were collected from clinically healthy adult (7–12 years of age) non-pregnant female camels during the breeding season at a local abattoir. Follicles (0.7–1.8 cm in diameter) did not coexist with a CL were classified as follicles F1 and their blood serum labeled F1S (n=12), follicles coexist with CL on the same ovary were labeled F2 and their blood serum labeled as F2S (n=9), and follicles with contralateral CL on the other ovary labeled F3 and their blood serum named F3S (n=11). Follicular fluid (FF) and sera were subjected to biochemical and hormonal analysis. Results revealed greater (P<0.05) concentrations of progesterone in the FF of F1 follicles and F1S serum than that found in F3 follicles and F3S serum. The concentrations of cortisone and T3 were higher (P<0.05) in FF of F1 and F3, and serum of F1S and F3S in comparison to FF of follicles type F2 and serum of F2S. The thyroxine and vitamin C concentrations in FF from F1 and F2 follicles, and serum of F1S and F2S were higher (P<0.01) in comparison with the FF from F3 and serum of F3S. The presence of CL might exert a local effect on hormonal composition of FF and could indirectly influence follicular development and oocyte quality. It is better to use oocytes originated from ovaries without a CL in in-vitro fertilisation (IVF) in dromedary camels.

OBESITY-RELATED ALTERATIONS IN PROTEIN EXPRESSION IN HUMAN FOLLICULAR FLUID

Obesity has a complex and significant impact on female reproduction. Prior research demonstrates that obesity is associated with decreased chances of conception, longer time-to-pregnancy, decreased response to medications in patients undergoing in vitro fertilization (IVF), and poorer IVF outcomes. In addition, obesity has been shown to directly alter the follicular microenvironment resulting in abnormal hormonal and metabolomic profiles. While proteomics has previously been employed to investigate follicular fluid in women with PCOS, there is a paucity of data investigating the impact of obesity in normo-ovulatory women. This study aimed to compare the proteomic composition of follicular fluid from normal vs. obese women without a history of PCOS or other ovulatory dysfunction undergoing IVF. Follicular fluid from 8 normal weight women (NW) and 8 women with obesity (OW) undergoing IVF was analyzed for proteomic content. Women undergoing IVF and without a history of PCOS, ovulatory dysfunction, endometriosis and diminished ovarian reserve were eligible for study inclusion. At the time of oocyte retrieval, follicular fluid from the 2nd and/or 3rd follicle from each ovary was collected and cryopreserved. Follicular fluid was subsequently thawed, and the most abundant proteins depleted using the ProteoMinerTM depletion kit (Biorad). Individual samples were then proteoylsed with Trypsin and labeled with a TMT 10-plex kit (ThermoFisher). Liquid chromatography mass-spectrometry (LC-MS) analysis was then performed through the University of Michigan proteomics core. Proteome Discoverer (v2.4) was used for data analysis. The ratio of protein abundance from obese/normal weight samples was calculated and the error rate corrected by utilization of the Benjamini-Hochberg method. Statistical significance was determined by a p<0.05. Western blotting was performed for validation of a subset of proteins. The mean age of NW vs OW was similar (32.88 vs 32.63 yrs). The mean BMI of NW was 21.1 kg/m2 compared with a BMI of 37.05kg/m2 in OW. A total of 1174 proteins were identified with >= 2 peptides present. 25 proteins were found to be significantly altered in the follicular fluid from women with obesity. 19 of these proteins were upregulated while 6 were downregulated. Notably, C-reactive protein was 11-fold higher in the follicular fluid from OW compared with NW (p<0.0001). Confirmatory western blots were consistent with the LC-MS findings. In this study we utilized high-resolution LC-MS to identify 1174 proteins in human follicular fluid. Further we found significant alterations in the proteomic composition of follicular fluid from women with obesity vs normal weight women without a history of ovulatory dysfunction. Future studies are needed to validate these findings in larger populations of patients and to further explore the mechanisms of these altered proteins in the pathophysiology of obesity.

P–196 Bacterial influence on oocyte quality - the secret of a successful fertilization

Abstract Study question Is there a difference in bacterial composition of follicular fluid between oocytes developing a good quality blastocyst and oocytes that fail fertilization? Summary answer Follicular fluids of oocytes failing fertilization show a different bacterial profile compared to follicular fluids of oocytes that were successfully fertilized. What is known already The presence of pathogens in the female reproductive tract has been intensively investigated. Lactobacillus species are mainly associated with a healthy genital tract and good prognosis for a successful pregnancy. Studies of the bacterial composition of follicular fluids have been mainly undertaken in women participating in reproductive medicine treatment because of the nature to obtain the specimen. In most studies follicular fluids have been pooled for analysis. Information on separately collected follicular fluids is still rare. We hypothesized that the composition of bacteria within follicular fluids is responsible for the success of the fertilization process. Study design, size, duration The study was designed and conducted at the Kinderwunsch Institut Schenk GmbH (Dobl, Austria) together with DNA-Technology. Follicular fluids from 46 patients undergoing IVF (in vitro fertilization) and ICSI (intracytoplasmic sperm injection) treatment were included and analyzed. Participants/materials, setting, methods Follicular fluids from 46 patients were collected separately. 2 follicular fluids from each patient were screened for common bacteria of the genital tract. One from an oocyte developing a good quality blastocyst and one displaying fertilization failures. Samples were analyzed for bacterial composition using the Femoflor16 (DNA-Technology). Main results and the role of chance Quantitative analysis revealed a higher total bacteria mass in follicles from oocytes that failed fertilization. Furthermore, Lactobacillus were not present in those follicles compared to good blastocyst follicles. In addition, Chlamydia trachomatis was found mainly in follicular fluid of not fertilized oocytes together with Eubacterium, Gardnarella and Trichomonas species. Interestingly, a trend of elevated levels of Ureaplasma species in follicular fluids of oocytes developing good quality blastocysts was observed. Limitations, reasons for caution Contamination of follicular fluids due to the procedure of oocyte pick up and follicular fluid retrieval cannot be completely excluded. Results should be confirmed with a higher sample size. Wider implications of the findings: We assume that different bacterial compositions in follicular fluids are responsible for the destiny of the oocyte. It is tempting to speculate that bacterial analysis of follicular fluids may be beneficial to select to best oocytes in future IVF/ICSI treatments. Trial registration number Not applicable

Effects of a dietary supplement enriched in palmitoleic acid on fatty acid composition of follicular fluid, granulosa cell metabolism, and oocyte developmental capacity in early lactation dairy cows

In high-yielding dairy cows, some fertility traits can be influenced by the fatty acid (FA) composition of the follicular fluid during early lactation. The first objective of the current study was to evaluate the potential of dietary supplements enriched in specific FA to influence the FA composition of follicular fluid lipid classes in early lactation dairy cows. The second objective was to determine the influence of the resulting follicular fluid FA composition on the folliculogenesis, lipid and energy metabolism of granulosa cells, as well as oocyte quality and embryo development. Twenty Holstein multiparous cows in late gestation were randomly assigned to 200 g/d of FA supplements enriched in (1) palmitic acid (control treatment; 82% 16:0; PA) in the rumen or (2) palmitoleic acid (sea buckthorn oil; 27% cis-9 16:1, 28% 16:0, 22% cis-9 18:1, and 11% cis-9,cis-12 18:2; SBT) in the abomasum. The treatment period ranged from 20 ± 5 d precalving to 67 ± 2 d postcalving. Cumulus-oocyte complexes, granulosa cells, and follicular fluid were recovered from 2 sequential sessions of ovum pick-up (OPU-1 and OPU-2) at 46 and 67 ± 2 d postcalving (mean ± standard deviation). On the same days, blood samples were collected. Milk performance was recorded, and feed and milk samples were collected from d 8 to 10 ± 3 (onset of lactation), d 35 to 37 ± 2 (before OPU-1), and d 63 to 65 ± 2 (before OPU-2). Treatments did not affect milk yield or fat concentration throughout the experimental trial. Compared with PA, SBT increased the cis-9 16:1 concentration in milk fat, in plasma esterified lipid classes (phospholipids, cholesterol esters, and triacylglycerols), and in follicular fluid phospholipids and cholesterol esters at OPU-1. Abundance of mRNA for stearoyl-CoA desaturase 1 and 5, and perilipin 2 in granulosa cells was not different between treatments, but an increase in the level of stearoyl-CoA desaturase 5 was observed between the 2 OPU periods. Treatments did not affect oocyte quality and developmental capacity or embryo lipid metabolism when cultivated in vitro. These results suggest that limited modifications in the FA composition of the oocyte microenvironment via dietary lipid supplements enriched in specific FA had no major effects on granulosa cell metabolism and oocyte developmental capacity in early lactation cows.

Comparative Analysis of Biochemical Constituents between Ovarian Follicular Fluid and Serum in Ganjam Goat (Capra hircus)

Background: Follicular fluid is in part exudates of serum and is also partially composed of locally produced substances, which are related to the metabolic activity of the follicular cells. The knowledge of the biochemical composition of follicular fluid can provide useful information about the requirements, growth and maturation of oocyte and which may be used as a provisional guide for formulating suitable culture media for in vitro cell culture and maturation in a particular species. Methods: Follicular fluid were aspirated from small follicles ( less than 3 mm; SF), medium follicles (3-5 mm; MF) and large follicles ( greater than 5 mm; LF). Blood samples were also collected for extraction of serum. Follicular fluid and serum samples were analyzed for biochemical constituents. Conclusion: It can be concluded that some biochemical constituents of follicular fluid such as glucose, cholesterol, total protein and globulin were higher in large follicles compared to small follicles. Moreover, serum levels of most of biochemical constituents were higher than their levels in different sizes of follicular fluid. This present investigation of comparative analysis of biochemical constituents between serum and ovarian follicles in Ganjam goat of Odisha will provide clues to researchers who work on reproductive physiology in different breeds of goat.

The Fatty Acid Composition of Human Follicular Fluid Is Altered by a 6‐Week Dietary Intervention That Includes Marine Omega‐3 Fatty Acids

The fatty acid composition of human follicular fluid is important for oocyte development and for pregnancy following in vitro fertilization (IVF). This study investigated whether a dietary intervention that included an increase in marine omega-3 fatty acids, olive oil and vitamin D alters the fatty acid composition of human follicular fluid. The association of lifestyle factors with follicular fluid fatty acid composition was also investigated. Fifty-five couples awaiting IVF were randomized to receive the 6-week treatment intervention of olive oil for cooking, an olive oil-based spread, and a daily supplement drink enriched with vitamin D and the marine omega-3 fatty acids docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) and 56 couples were randomized to receive placebo equivalents. Dietary questionnaires were completed, and samples of blood were taken before and after the intervention. Follicular fluid was collected at oocyte retrieval and the fatty acid profile assessed using gas chromatography. In the control group, individual fatty acids in red blood cells and follicular fluid were significantly correlated. Furthermore, a healthier diet was associated with a lower percentage of follicular fluid arachidonic acid. The follicular fluid of women in the treatment group contained significantly higher amounts of EPA and DHA compared to the control group, while the omega-6 fatty acids linoleic, γ-linolenic, dihomo-γ-linolenic, and arachidonic were lower. This is the first report of a dietary intervention altering the fatty acid composition of follicular fluid in humans. Further research is required to determine whether this intervention improves oocyte quality.

Relationship between the metabolic and lipid profile in follicular fluid of women undergoing in vitro fertilization.

Among the follicular fluid (FF) components promoting the development of the oocyte are included glycoproteins, several fatty acids, and steroid hormones synthesized by the dominant follicle. For this, the analysis of the metabolites present in FF can determine the quality of the oocyte. FF composition is in part determined by local follicular metabolic processes and in part a plasma transudate. Since the causes of impaired fertility may be due to a metabolic imbalance, metabolomics is useful to identify low molecular weight metabolites. Oxidative stress is involved in human infertility and the use of metabolomics can be crucial to identify which other metabolites besides reactive oxygen species are involved in oxidative stress correlated to infertility. To obtain new information on the study of signaling molecules in FF, the knowledge of the lipid content will be important to improve information on the understanding of follicular development. The objective of this study is to identify (a) a metabolic profile and a lipid profile of FF in women undergoing in vitro fertilization and (b) to correlate the previous information obtained regarding adiponectin and oxidative stress with the metabolic and lipid profile obtained in the present study. As result, we found an increase in oxidative stress due to both an increase of androgens and an accumulation of lipids in the follicular environment and we suggest that this might be one of the causes of reduced fertility.

AGE-EFFECT ON THE FATTY ACID AND LIPIDOMIC PROFILE OF THE FOLLICULAR FLUID DURING IN-VITRO FERTILIZATION CYCLES

Compelling evidence suggests that the metabolic profile and lipidomic composition of the human follicular fluid (FF) may play a role in reproductive outcomes. However, there are gaps in current knowledge regarding the follicular environment of aging, which is the main factor affecting female reproductive capacity. Thus, we aimed to evaluate if there is any variability in the fatty acid and lipidomic composition of the FF between oocyte donors and infertile patients with advanced age.

Metabolic changes in follicular fluids of patients treated with recombinant versus urinary human chorionic gonadotropin for triggering ovulation in assisted reproductive technologies: a metabolomics pilot study.

The main goal of this retrospective cohort study is the assessment of the effects of administration of recombinant-hCG (r-hCG) versus urinary-hCG (u-hCG) on follicular fluid (FF) composition of women who underwent in vitro fertilization (IVF) treatments. We selected 70 patients with infertility attributable to tubal diseases, unexplained infertility, and male factor. Metabolomics analysis of their FFs was performed by 1H nuclear magnetic resonance (1H NMR) spectroscopy in combination with multivariate analysis to interpret the spectral data. Univariate statistical analysis was applied to investigate the possible correlations between clinical parameters and between clinical parameters and metabolites identified by NMR. According to the type of hCG used, significant differences were detected in FFs of women with male factor and unexplained infertility, both in qualitative and quantitative terms, for some metabolites as cholesterol, citrate, creatine, β-hydroxybutyrate, glycerol, lipids, amino acids (Glu, Gln, His, Val, Lys) and glucose. No significant difference was observed in women with tubal diseases. Besides, the number of MII oocytes in the u-hCG-treated groups correlates positively with glutamate in tubal disease and with glycerol in unexplained infertility. In the r-hCG-treated groups, the number of MII oocytes correlates positively with lipid in tubal disease, positively with citrate and negatively with glucose in male infertility. Metabolite composition of FF changes according to different type of hCG treatment and this can be related to oocyte development and subsequent outcome. According to the data of this study, different types of hCG should be used in relation to the diagnosis of infertility to obtain better results in inducing oocyte maturation in women undergoing IVF.

The follicular microenvironment in low (++) and high (I+B+) ovulation rate ewes.

Ewes with single copy mutations in GDF9, BMP15 or BMPR1B have smaller preovulatory follicles containing fewer granulosa cells (GC), while developmental competency of the oocyte appears to be maintained. We hypothesised that similarities and/or differences in follicular maturation events between WT (++) ewes and mutant ewes with single copy mutations in BMP15 and BMPR1B (I+B+) are key to the attainment of oocyte developmental competency and for increasing ovulation rate (OR) without compromising oocyte quality. Developmental competency of oocytes from I+B+ animals was confirmed following embryo transfer to recipient ewes. The microenvironment of both growing and presumptive preovulatory (PPOV) follicles from ++ and I+B+ ewes was investigated. When grouped according to gonadotropin-responsiveness, PPOV follicles from I+B+ ewes had smaller mean diameters with fewer GC than equivalent follicles in ++ ewes (OR = 4.4 ± 0.7 and 1.7 ± 0.2, respectively; P < 0.001). Functional differences between these genotypes included differential gonadotropin-responsiveness of GC, follicular fluid composition and expression levels of cumulus cell-derived VCAN, PGR, EREG and BMPR2 genes. A unique microenvironment was characterised in I+B+ follicles as they underwent maturation. Our evidence suggests that GC were less metabolically active, resulting in increased follicular fluid concentrations of amino acids and metabolic substrates, potentially protecting the oocyte from ROS. Normal expression levels of key genes linked to oocyte quality and embryo survival in I+B+ follicles support the successful lambing percentage of transferred I+B+ oocytes. In conclusion, these I+B+ oocytes develop normally, despite radical changes in follicular size and GC number induced by these combined heterozygous mutations.

Follicular fluid and assisted reproductive technology programs outcomes (literature review)

The review presents data on metabolites in the follicular fluid (FF) from the perspective of reproductive medicine and their use in order to predict outcomes of assisted reproductive technology (ART) programs. It considers various components of this biological medium (hormones, lipids, melatonin, etc.) with an assessment of their predictive value in prognosis of the effectiveness of in vitro fertilization (IVF) programs. The data on experimental directions in this field and the prospects for their use in clinical practice are presented. The article emphasizes that the growing clinical need and the unsolved problem of increasing the effectiveness of ART programs determine the need for further studies of the FF composition.&#x0D; Materials and methods. The review includes data related to this topic from foreign and Russian articles found in PubMed which were published in recent years.&#x0D; Results. Given the established fact of a direct effect of FF composition on growth and maturation of oocytes, and further, on the fertilization process, various FF metabolites are actively investigated as non-invasive markers of quality of oocytes/embryos. The article provides data on the experimental directions in this field and the prospects for their use in clinical practice. However, clinical studies of a relation between various FF metabolites levels and outcomes of IVF programs are contradictory.&#x0D; Conclusion. Owing large economic cost for treatment of infertility with IVF, there is need for expansion and intensification of studies to identify and use reliable predictors in prognosis of ART programs outcomes.