Outcomes of assisted reproduction treatment after dopamine agonist -cabergoline- for prevention of ovarian hyper stimulation syndrome.

BACKGROUND: Release of vascular endothelial growth factor (VEGF) by ovaries in response to HCG administration is one of the main mechanisms of ovarian hyper stimulation syndrome. Since Dopamine/dopamine receptor2 (Dp-r2) pathway activity -mediated by VEGF/ Vascular endothelial growth factor receptor 2 (VEGFR- 2) signaling-, is associated with angiogenic events, dopamine agonists were used for the management of severe forms of OHSS. In order to assess the effects of Cabergoline on angiogenesis in the human endometrium, and subsequently its impacts on the implantation rate this study was conducted. METHODS: This historical cohort study was conducted based on existing data of 115 patients (20-40 years) whom underwent assisted reproductive treatment (ART) and with a high probability for developing OHSS between March 2007 and September 2008. Forty five cases received Cabergoline were compared to 70 control subjects. The statistical methods used were: Unpaired t-test for continuous variables and the chi-square test (or Fisher's exact test if required) for categorical variables. RESULTS: None of the patients (treatment or control group) developed OHSS. The etiologies of infertility and administration of GnRH agonist or antagonist protocols were similar in two groups (p>0.2). Number of transferred embryos and zygote intra-fallopian transfer (ZIFT) did not differ between the two groups (p≥0.06). Implantation rate in treatment (3.1%) and control (6.6%) subjects was similar (p=0.4). No significant difference was observed in fertilization rate, chemical, clinical and ongoing pregnancies between the two groups (p>0.5). CONCLUSION: Cabergoline can be safely administered in ART protocols to prevent OHSS, without compromising ART outcomes.

Embryo transfer in days 2 to 4 following intracytoplasmic sperm injection: a prospective cohort study.

BACKGROUND: Improvement of assisted reproductive technique (ART) results in higher pregnancy rates from positive Beta HCG to take home baby statistics. Despite developments in culture media allowing blastocyst stage transfer, some centers apply second, third and sometimes fourth day post injection for embryo transfer. This study aimed to compare their reproductive outcomes. METHODS: This prospective cohort study conducted on 218 infertile couples with at least 4 oocytes retrieved and 2 good quality embryos. They were divided consecutively into 2nd (ET2) or 3rd (ET3) day embryo transfer. Some patients experienced 4rd (ET4) day embryo transfer due to weekend reasons, so we included them in our comparison as well. There were 98, 97 and 23 patients in the aforementioned groups, respectively. Reproductive and pregnancy outcomes were evaluated by Chi square and t-test with the significance level set at α=0.05. RESULTS: Totally, 73 patients (33%) had positive beta HCG and 39.7 percent of them (n=29) experienced pregnancy loss. Positive Beta HCG was detected in 31(31.6%) of ET2 patients, 38 (39.2%) of ET3 patients and 4 (17.4%) of ET4 group. Abortion or pregnancy loss was reported in 9 (29%) of ET2 patients, 18 (47.4%) of ET3 patients and 2 (50%) of ET4 group. CONCLUSION: Our study demonstrated that there may be a higher pregnancy as well as higher abortion in day 3 embryo transfer.

Hysteroscopy in patients with repeated implantation failure improves the outcome of assisted reproductive technology in fresh and frozen cycles.

AIM: Repeated implantation failure (RIF) is still a problem for many patients and their physicians. Some interventions have been practiced to overcome the problem; one is uterine cavity assessment before assisted reproductive technology (ART) cycles. This study aimed to evaluate the effect of hysteroscopy in women experiencing recurrent implantation failure with apparently normal uterine cavity before assisted reproductive techniques. MATERIAL AND METHODS: This was a cohort study with historical controls conducted in a university hospital. A total of 353 women with RIF undergoing ART with normal hysterosalpingography and transvaginal ultrasound were evaluated. The intervention group underwent hysteroscopy with a rigid, 30°, 4-mm hysteroscope in the menstrual cycle just before ART; in the control group hysteroscopy was not performed. Basal characteristics, stimulation parameters and pregnancy rates were compared between the two groups. RESULTS: Chemical pregnancy occurred in 58.5% of women in the hysteroscopy group versus 34.1% of control women (odds ratio [OR]: 2.7; 95% confidence interval [CI]: 1.7-4.2; P < 0.001). Clinical pregnancy occurred in 50.7% and 30.3% of women in the hysteroscopy and control groups, respectively (OR: 2.4; 95%CI: 1.5-3.7; P < 0.001). Delivery rate was 35.5% in hysteroscopy women and 21.1% in control women (OR: 1.9; 95%CI: 1.2-3.1; P = 0.008). The results of hysteroscopy were normal in 103 women (72.5%), and they revealed inflammation in 22 (15.5%), polyp in 16 (11.3%) and Asherman syndrome in one patient (0.7%). CONCLUSIONS: Hysteroscopy in the menstrual cycle before ovarian stimulation in fresh cycles and before endometrial preparation in frozen thawed cycles in women experiencing recurrent implantation failure with apparently normal uterine cavity significantly increases the pregnancy rates in fresh and frozen cycles, respectively.

Genome instability in AZFc region on Y chromosome in leukocytes of fertile and infertile individuals following exposure to gamma radiation.

PURPOSE: Men are exposed to various doses of ionizing radiation due to living in regions with high natural background radiation, accidentally, occupationally or for cancer treatment. To study genomic instability of AZFc region to gamma radiation, blood samples from normal, oligozoospermia, and azoospermia individuals were irradiated by a Co-60 source. METHODS: Irradiated cells were kept for 48 h in order to repair initial DNA damages. Real time PCR was performed for three markers (SY 1206, SY 1197, SY 579) for testing copy number variation before and after irradiation. Copy number variations were compared by calculation of cycle threshold comparative method. RESULTS: Copy number variations of studied markers in AZFc region (microdeletion and duplication) in all samples after exposure to radiation increased with a dose dependent fashion. The frequency of instability was significantly higher in samples from infertile men in comparison with fertile ones (p < 0.001). CONCLUSION: No significant difference was seen between the two infertile groups (p > 0.05). This observation might be a possible explanation for induction of azoospermia and oligozoospermia after radiotherapy. Increased frequency of induced microdeletion and duplication in infertile men compared with normal might be attributed to the deficiency in repair systems and the genetic factors involved in incomplete spermatogenesis of infertile men.

Estradiol supplementation during the luteal phase in poor responder patients undergoing in vitro fertilization: a randomized clinical trial.

PURPOSE: This study was designed to evaluate the effects of adding Estradiol (E2) supplementation to progesterone (P) on improvement of pregnancy outcomes in poor responder patients who underwent in vitro fertilization (IVF). METHODS: In a prospective randomized clinical trial, 118 poor responder patients, older than 38 years without contraindications of estradiol consumption from Infertility clinic of a university hospital were randomly divided (by computerized software) into two groups. Control group (59 patients) received only P and intervention group (59 patients) received P and E2 (4 mg/d). Supplementation was done with 4 mg E2 in the luteal phase. Fertilization rate, implantation rate, biochemical and clinical pregnancy rates, abortion rate, ongoing pregnancy, multiple pregnancy and ectopic pregnancy rates were documented for those who completed the study protocol in each group (per protocol analysis) and compared between groups. RESULT: Fifty five patients in control group and 53 patients in intervention group successfully completed the study protocol. Treatment outcomes were not significantly different between two groups. CONCLUSION: For poor responder women who underwent IVF, addition of E2 to P supplementation could not significantly improve pregnancy outcomes.

The effectiveness of cabergoline for the prevention of ovarian hyperstimulation syndrome.

Type 2 receptors for vascular endothelial growth factor are believed to be involved in the pathophysiology of ovarian hyperstimulation syndrome (OHSS). The objective of this study was to examine the preventive effects of cabergoline on OHSS and its complications. The study is a non randomized clinical trial conducted in 2006-2008 on 75 patients, which were at risk of OHSS and underwent assisted reproductive techniques. The diagnosis and severity of OHSS were determined using standard criteria. The study included an intervention and a control group. The intervention group comprised of 50 women at risk of OHSS, who were treated with cabergoline (1 mg every other day for 8 days) commencing from the day of ovum pick up. The control group comprised of 25 historical cases, which were similar to the case group. The latter group did not receive cabergoline, and their OHSS, if occurred, were managed conservatively after hospital admission. The rates of OHSS, baseline characteristics, ovarian stimulation parameters, and pregnancy occurrence were compared. There was no significant difference between baseline characteristics or ovarian stimulation parameters form the two groups. The incidence of OHSS in the cabergoline-treated group, was significantly (P=0.01) lower than that in the control group (12% vs 36%). Embryo freezing was significantly (P=0.001) lower in the control group, but cycle cancellation was significantly (0.03) lower in the cabergoline group. The findings of the study indicate that cabergoline reduces the incidence of OHSS, and is not associated with adverse effects on pregnancy.

Aqueous Origanum vulgare Extract Improves the Quality of Cryopreserved Human Spermatozoa Through Its Antioxidant Effects.

Excessive production of reactive oxygen species (ROS) during semen cryopreservation can induce structural and functional changes in spermatozoa. It is well known that antioxidants can mitigate the effect of ROS. Moreover, the application of antioxidants in freezing media is an appropriate strategy for protecting spermatozoa against deleterious effects of ROS during the cryopreservation process. As an example, oregano is a medicinal plant with important activities, with antiseptic, antibacterial, antithrombotic, and antioxidant properties. This study aimed at evaluating the antioxidant effects of oregano extract on cryopreserved human spermatozoa. In the first phase, 13 semen samples with different concentrations of oregano extract (0.0, 50, 100, 150, 300, and 500 µg/mL) were cryopreserved to achieve an optimal dose of oregano extract. Then, motility, viability, and plasma membrane integrity were evaluated. In the second phase, 20 samples were cryopreserved in freezing media supplemented with or without the optimal concentration of oregano (100 µg/mL). After thawing, motility, the levels of ROS, lipid peroxidation, and translocation of phosphatidylserine (PS) were evaluated. The results showed that 100 µg/mL oregano extract significantly increased the total motility in frozen-thawed spermatozoa in comparison with the control group (28.2 ± 4.3 vs. 42.4 ± 1.6, p < 0.05). This concentration significantly decreased the percentage of 2',7'-dichlorofluorescein-positive cells (25.53 ± 1.2 vs. 21.48 ± 1.2) and the malondialdehyde level (4.25 ± 0.7 vs. 0.82 ± 0.4 µM) (p < 0.05). In the oregano group, the percentage of vital spermatozoa without PS externalization was significantly higher than that in the control group (25.88 ± 1.6 vs. 16.8 ± 1.9, p < 0.001), while the percentage of dead spermatozoa with PS externalization spermatozoa was significantly lower than that in the control group (51.65 ± 1.4 vs. 60.36 ± 1.9, p < 0.05). In general, the addition of oregano extract to sperm freezing extender has protective effects against oxidative stress and apoptosis.

The protein-creatinine ratio in spot morning urine samples and 24-h urinary protein excretion in patients with systemic lupus erythematosus.

A 24-h urinary protein is a standard way to diagnose lupus nephritis. Assessment of protein-creatinine (Pr-Cr) ratio in morning spot urine is a valuable method in diabetic patients but not use in systemic lupus erythematous (SLE) patients routinely. In this study Pr-Cr ratio in spot urine was compare with 24-h urine protein; if they have valuable correlation we can use this test instead of 24-h urinary protein. The aim of this study was to evaluate the correlation of spot urine Pr-Cr ratio for prediction of significant proteinuria (>or=300 mg/24 h) in patients with SLE. A cross-section study was conducted in 74 hospitalized women with SLE. The correlation between Pr-Cr in first morning urine specimens and urinary protein excretion in 24-h collections were analyzed. Correlation between Pr-Cr ratio in spot morning urine specimens and urinary protein excretion in 24-h collections was significant (P < 0.0001, r = 0.83). A high correlation and precision of agreement were demonstrated between the two methods of assessment proteinuria in lupus patients. The difference between the two methods was less than the biological variability in the protein excretion and its measurement, enabling the methods to be used interchangeably creatinine ratio in spot morning urine samples is a precise indicator of proteinuria in patients with lupus nephritis and represents a simple and inexpensive procedure in establishing severity of proteinuria in patients with SLE.

High frequency of de novo DAZ microdeletion in sperm nuclei of subfertile men: possible involvement of genome instability in idiopathic male infertility.

The occurrence and diagnosis of Y-chromosome microdeletions, specifically deletions of the DAZ (Deleted in Azoospermia) genes are an important issue in male infertility. Screening Y chromosome microdeletion is mainly done using polymerase chain reaction (PCR) on blood leukocytes. However, there is some evidence indicating that presence of DAZ in somatic cells might not be indicative of its presence in the germ cell lineage. Therefore, a total of 130 men with poor semen quality were examined for presence of DAZ microdeletion in their leukocytes. From these, sperm from 40 randomly selected men with no DAZ microdeletions in their leukocytes (n = 10 oligozoospermia; n = 10 asthenozoospermia; n = 10 oligoasthenozoospermia; and n = 10 near-azoospermia) were were compared to sperm from men of normal semen quality (n = 10) using combined primed in situ labelling and fluorescent in situ hybridization (PRINS-FISH) technique as well as screening for sex chromosome aneuploidy. There was an increased frequency of DAZ microdeletion in blood samples from oligozoospermic (5%) (p < 0.05) and near azoospermic patients (14%) (p < 0.01). A high frequency of DAZ microdeletion was observed in the sperm of patients with no DAZ microdeletion in their leukocytes compared to control (p < 0.01). The frequency of sex chromosome aneuploidy also increased, correlating with the severity of infertility in the studied groups. A similar result was observed for sex chromosome aneuploidy. The results might be indicative of DAZ microdeletion induction during spermatogenesis.

Endometrial Preparation for Women Undergoing Embryo Transfer Frozen-Thawed Embryo Transfer With and Without Pretreatment With Gonadotropin Releasing Hormone Agonists.

Objective: To evaluate the efficacy of endometrial preparation by exogenous steroids, with and without pretreatment by the use of GnRH agonist. Materials and methods: This randomized interventional study was conducted on 100 women who underwent a FTET that were randomly assigned to receive GnRH agonist (buserelin) in the luteal phase or no receive this medication. In both groups endometrial preparation was achieved by the use of estradiol valerate pill started from the second day of the menstruation and used every day, with an initial dose of 2mg/d and every 3 days increased to 4 mg/d and 6 mg/d, respectively. Endometrial thickness was evaluated by vaginal ultrasound. Forty eight hours after beginning of progesterone administration 2 to 3 embryos surviving in freezing procedure were transferred. Results: the two groups were similar in mean age, body mass index, duration of infertility, type of infertility, number of embryos transferred and endometrial thickness on the day of beginning progesterone therapy. Comparing outcome of FTET between the two groups scheduled for receiving GnRH agonist showed no significant difference in the rate of implantation (6.7% versus 10.0%), the rate of chemical pregnancy (21.7% versus 22.5%), clinical pregnancy rate (15.0% versus 17.5%), and also ongoing pregnancy (13.3% versus 12.5%). Conclusion: Endometrial preparation for FTET using GnRH agonists appears to be as effective as FTET without administrating these agonists.

Can letrozole plus HMG protocol improve pregnancy outcomes in frozen-thawed embryo transfer? An RCT.

BACKGROUND: There are different methods in endometrial preparation for frozen-thawed embryo transfer (FET). OBJECTIVE: The purpose of this study was to compare the live birth rate in the artificial FET protocol (estradiol/ progesterone with GnRH-agonist) with stimulated cycle FET protocol (letrozole plus HMG). MATERIALS AND METHODS: This randomized clinical trial included 100 women (18-42 years) randomly assigned to two groups based on Bernoulli distribution. Group I received GnRH agonist [Bucerelin, 500µg subcutaneously] from the previous midlutea lcycle, Then estradiol valerat [2 mg/ daily orally] was started on the second day and was increased until the observation of 8mm endometrial thickness. Finally progesterone [Cyclogest, 800 mg, vaginally] was started. Group II received letrozole on the second day of the cycle for five days, then HMG 75 IU was injected on the7th day. After observing [18 mm folliclhCG10000 IU was injected for ovulation induction. Trans cervical embryo transfer was performed in two groups. The main outcome was the live birth rate. The rate of live birth, implantation, chemical, and clinical pregnancy, abortion, cancellation and endometrial thickness were compared between two groups. RESULTS: Implantation rate was significantly higher in group I. Live birth rate was slightly increased in group I without significant difference (30% vs. 26%). The rate of chemical and clinical pregnancy was similar in two groups. The abortion rate was lower in letrozole protocol but the difference was not statistically significant. The mean endometrial thickness was not different between two groups. CONCLUSION: Letrozole plus HMG method cannot improve pregnancy outcomes in frozen-thawed embryo transfer but it has only one injection compare to daily injections in artificial method.

High frequency of microdeletion in TTY2 gene family in peripheral blood leukocytes of non-obstructive azoospermia patients.

About 10-15% of non-obstructive azoospermia (NOA) patients show AZFc microdeletion in their blood leukocytes. However, if AZF genes were involved in impaired spermatogenesis, a higher frequency of chromosomal microdeletions was expected. In this study the frequency of AZFc microdeletion was compared with TTY2 gene family, i.e., TTY2A2A and TTY2A12A in blood leukocytes of NOA patients and normal fertile control. In the present study 30 normal fertile individuals with mean age of 35.0 ± 6.0 and 30 NOA patients with mean age of 34.0 ± 7.0 were screened for microdeletion of TTY2L2A and TTY2L12A at Yq11 and Yp11 respectively and sequence-tagged site (STS) markers for AZFc gene using multiplex PCR technique. At the first step karyotyping was done for all subjects using standard G-banding technique to identify patients with normal karyotype as well as non-affected normal controls for molecular analysis. Results showed no AZFc microdeletion in normal and NAO patients whereas one TTY2L2A microdeletion in normal control (3.3%) and 4 in NOA (13.3%) was observed (p < 0.05). However our data indicated that 6 of 30 NOA patients (20%) showed TTY2L12A microdeletion whereas there was no observed microdeletion in normal control (p < 0.01). Results indicate that the studied genes might be involved in impaired spermatogenesis more effective than the routinely screened AZF genes in infertile men. Therefore, screening these genes along with AZF genes might be valuable for infertile patients. The reason why these genes are deleted from Y chromosome is not known but might be associated with genomic instability induced by environmental physico-chemical genotoxic agents.

Nerve growth factor in human semen: Effect of nerve growth factor on the normozoospermic men during cryopreservation process.

OBJECTIVES: Although routinely applied in assisted reproductive technology, human sperm cryopreservation is not a completely successful procedure. Adverse effects of cryopreservation on the fertilization capacity, motility, morphology, and viability of spermatozoa have been proven; cryopreservation has also shown a role in sperm DNA fragmentation and infertility. The post-thaw survival of spermatozoa improved after addition of supplementation of antioxidant molecules to freezing media. Nerve growth factor (NGF) as one of the prosurvival substances has gained great attention in recent years. The aim of this study was the usage of NGF as prosurvival factor after cryopreservation process of human semen samples to assess the motility and viability of sperm, nitric oxide (NO) concentration, and DNA fragmentation in normozoospermic men. MATERIALS AND METHODS: Semen samples were collected from 25 normozoospermic men and were divided into fresh semen samples as control group, frozen-thawed semen samples without addition of exogenous NGF, and three groups of semen samples cryopreserved with addition of exogenous NGF (0.5, 1, and 5 ng/ml) in freezing medium. Viability was assessed by eosin-negrosin staining technique. Motility was evaluated with inverted microscope. NO concentration and apoptosis content were measured with flow cytometry. RESULTS: Results showed that exogenous NGF at 0.5 ng/ml could significantly (P-value <0.05) influence viability, motility, nitric oxide, and DNA fragmentation content. CONCLUSION: Exogenous NGF as cryoprotectant improved sperm viability and motility, increased intracellular NO concentration, and decreased apoptosis content in normal human spermatozoa.

Prediction of assisted reproductive technique outcome in elevated early follicular phase follicle stimulating hormone with Mullerian inhibiting substance level.

BACKGROUND: Detection of best predictor of ovarian reserve in patients with temporarily or consistently elevated early follicular phase serum levels of FSH is one of the most important goals in assisted reproductive technique (ART). OBJECTIVE: To evaluate whether high level of anti-mullerian hormone level is related to success of ART in patients with temporarily or consistently elevated early follicular phase serum levels of FSH. MATERIALS AND METHODS: Sixty three women underwent intracytoplasmic sperm injection (ICSI) with GnRH-agonist long protocol or intrauterine insemination (IUI) in a prospective cohort study. FSH, inhibin B and anti-Mullerian hormone (AMH) levels were measured in these women whom were divided to three groups (persistently elevated FSH, variably elevated FSH and, normal FSH level). Basal characteristics, stimulation parameters, and pregnancy occurrence were evaluated. RESULTS: AMH was significantly higher in women with persistently elevated early follicular phase FSH achieving pregnancy. Women with normal FSH did not have significant difference in AMH level between conceived and non conceived cycles. Women with only one elevated early follicular phase FSH achieving pregnancy did not have significant difference in AMH level with non pregnant women. Response to gonadotropin stimulation, recommendation to oocyte donation significantly differed between the groups. CONCLUSION: This study has demonstrated that relatively young women with persistently or intermittently elevated day 3 FSH levels have diminished ovarian reserve and lower ART success. However, in women whose FSH levels were constantly elevated, AMH (not inhibin B) concentrations were significantly higher in ART cycles resulting in pregnancy. Therefore, AMH level is a good predictor of ART outcome in patients with elevated early follicular phase serum levels of FSH.

Anti-mullerian hormone as a predictive factor in assisted reproductive technique of polycystic ovary syndrome patients.

This study aimed to assess the relationship between the serum levels of anti-mullerian hormone (AMH) and other hormonal markers and results of assisted reproductive techniques (ART) in polycystic ovary syndrome (PCOS) patients. This cohort study was conducted on 60 PCOS patients who were candidates for assisted reproductive techniques. In all patients the serum levels of AMH, follicle stimulating hormone (FSH) and luteinizing hormone (LH), estradiol (E2), free testosterone (fT), testosterone (T) and inhibin B were measured in the 3(rd) day of menstrual cycle. The relationship between serum level of measured hormonal markers with retrieved oocytes, mature oocytes, the number of transferred fetus and pregnancy rate were assessed. The cut-off value for the serum level of AMH and retrieved oocytes were determined. There was a significant direct correlation between the serum mullerian inhibiting substance (MIS) level with number of total picked up oocytes (r = 0.412), mature oocytes (r = 0.472) and embryo transfer (r = 0.291). There was a linear and significant correlation between inhibin B and fertilization (r = 0.283) Cut-off point for AMH level according to presence or absence of pregnancy was 4.8 ng/ml and it was not statistically significant (P = 0.655). Area under curve (AUC) was 0.543. Cut-off point for MIS according to picked up oocytes was 2.7 ng/ml with area under the curve (ROC curve) of 0.724 (CI = 0.591-0.831) (P = 0.002). Patients with PCOS who had AMH more than 2.7 ng/ml, the number of retrieved oocytes (6 or more) was higher than MIS/AMH <2.7 ng/ml (P = 0.002). As a marker of ovarian responsiveness to controlled ovarian hyperstimulation (COH) and despite a small sample size of our study, it is revealed that pretreatment MIS/AMH is highly associated with the number of mature oocytes retrieved during COH in PCOS women.

Predictive value of the level of vitamin D in follicular fluid on the outcome of assisted reproductive technology.

OBJECTIVE: To assess the correlation between the levels of vitamin D in follicular fluid and serum, and to determine whether the level of 25-hydroxyvitamin D (25OH-D) in follicular fluid of infertile women undergoing assisted reproductive technology (ART) is associated with the outcome. STUDY DESIGN: Eighty-two infertile women undergoing ART at an academic tertiary care centre were recruited for a prospective cohort study. Levels of 25OH-D in follicular fluid and serum were measured. Standardized regimens for pituitary downregulation and controlled ovarian hyperstimulation were employed. Patient and cycle parameters, and clinical pregnancy (defined as evidence of intra-uterine gestation sac plus heart rate on ultrasound) were determined. RESULTS: A significant correlation was found between the levels of vitamin D in follicular fluid and serum (r=0.767, p=0.001). The overall rates of chemical, clinical and ongoing pregnancy were 35.5% (n=29), 29.3% (n=24) and 23.2% (n=19), respectively. No significant difference was found in these pregnancy rates between the tertiles of 25OH-D level in follicular fluid (p=0.959, 0.995 and 0.604, respectively). The median serum level of vitamin D was 8.13 (range 5.37-13.62)ng/ml in the clinically pregnant group and 8.29 (range 5.93-21.23)ng/ml in the non-pregnant group (p=0.235). Interestingly, the median level of vitamin D in follicular fluid was 9.19 (range 5.25-19.51)ng/ml in the clinically pregnant group and 10.34 (range 5.89-29.69)ng/ml in the non-pregnant group (p=0.433). The fertilization rate decreased significantly and the implantation rate increased (not significantly) with increasing tertiles of 25OH-D level in follicular fluid. CONCLUSIONS: The level of 25OH-D in follicular fluid is reflective of body stores of vitamin D. Most subjects in this study were deficient in vitamin D, but this study found that vitamin D deficiency does not play a pivotal role in the outcome of ART.