Richard V. Grazi

Bleeding and spontaneous abortion after therapy for infertility

OBJECTIVE To determine the incidence of early-pregnancy bleeding and spontaneous abortion (SAB) after various treatments for infertility and to assess whether bleeding is predictive of SAB. DESIGN An historic cohort study of women who conceived after various treatments. SETTING Hospital-based private practice. PATIENT(S) We studied 418 patients in whom 500 consecutive clinical pregnancies occurred. INTERVENTION(S) Patients were grouped according to the method of conception: ovulation induction, IVF, and other. The latter category included interventions not requiring ovulation induction, such as surgery and insemination. A fourth group of subjects who conceived independently of treatment was used as the control. MAIN OUTCOME MEASURE(S) Bleeding and pregnancy outcome (SAB, ectopic pregnancy, or ongoing pregnancy). RESULT(S) Rates of SAB did not differ among the treatment groups. SAB occurred significantly more often after bleeding than when bleeding did not occur (30.8% versus 19.8%, respectively). Bleeding was predictive of SAB only in patients <35 years old (odds ratio 2.4). CONCLUSION(S) Infertile women who conceive after reproductive therapy are not at increased risk for SAB compared with women who conceive naturally. There appears to be no association between previous diagnosis or treatment and the occurrence of SAB in previously infertile women. Bleeding is associated with a twofold relative risk of SAB.

Random anti-Müllerian hormone (AMH) is a predictor of ovarian response in women with elevated baseline early follicular follicle-stimulating hormone levels

OBJECTIVE To investigate the utility of random anti-Müllerian hormone (AMH) in assessing ovarian response among women with diminished ovarian reserve (DOR) diagnosed by elevated early follicular-phase FSH levels. DESIGN Retrospective study. SETTING Academic and academically affiliated assisted reproductive technology (ART) programs. PATIENT(S) Seventy-three women undergoing ART with elevated early follicular FSH levels. INTERVENTION(S) None. MAIN OUTCOME MEASURE(S) Number of oocytes retrieved during ART cycle, number of day 3 embryos, and cycle cancellation and clinical pregnancy rates. RESULT(S) Random AMH levels were strongly correlated with the number of oocytes retrieved during an ART cycle among women with elevated FSH (r=0.55). Women with elevated FSH who had a random serum AMH level of 0.6 ng/mL or higher had twice the number of oocytes retrieved (11±1.3 vs. 5.6±0.6), a greater number of day 3 embryos (5.7±0.9 vs. 3±0.5), and approximately a third of the cycle cancellation rate (14% vs. 41%) compared with women with a random serum AMH below 0.6 ng/mL. The clinical pregnancy rate was also higher among women with a random serum AMH≥0.6 ng/mL (28% vs. 14%), however, the difference was not statistically significant. CONCLUSION(S) A random serum AMH level is useful in setting expectations for ART prognosis in women with elevated early follicular-phase serum FSH levels.

Elevated body mass index is associated with lower serum anti-mullerian hormone levels in infertile women with diminished ovarian reserve but not with normal ovarian reserve

OBJECTIVE To investigate the association between elevated body mass index (BMI) and ovarian reserve. DESIGN Cross-sectional study. SETTING Academic institutions. PATIENT(S) Two hundred ninety women with infertility. Diminished ovarian reserve (DOR) was defined as day 3 FSH>10 IU/L. INTERVENTION(S) None. MAIN OUTCOME MEASURE(S) Random serum antimullerian hormone (AMH) levels and number of oocytes retrieved during assisted reproductive technology cycle. RESULT(S) Increasing BMI was associated with lower random serum AMH levels in infertile women with DOR but not in women with normal ovarian reserve (NOR). Among women with DOR, mean random serum AMH levels were 33% lower in overweight and obese women compared with women with normal weight. The same association was not true for women with NOR. Out of 290 women evaluated for infertility, 109 women underwent a controlled ovarian hyperstimulation (COH)-IVF cycle. Women with elevated BMI and DOR had lower number of oocytes retrieved compared with women with normal BMI and DOR (6.4±4.3 vs. 9.4±6), an association that was not observed among women with NOR. CONCLUSION(S) Overweight and obese women with DOR as defined by high day 3 serum FSH levels have lower serum AMH levels and number of oocytes retrieved compared with nonobese women with DOR.

Laparoscopic excision of benign dermoid cysts with controlled intraoperative spillage

Due to the risk of aseptic peritonitis associated with intraoperative rupture of dermoid cysts, many surgeons are reluctant to remove the lesions laparoscopically. In our series, 12 dermoid cysts were removed laparoscopically. Intraoperative spillage occurred in all cases and was managed with copious lavage. None of the patients experienced postoperative morbidity associated with peritonitis, indicating that intraoperative spillage of dermoid cysts is not associated with morbidity as long as vigorous lavage is performed.

Vitamin D Supplementation Decreases TGF-β1 Bioavailability in PCOS: A Randomized Placebo-Controlled Trial

CONTEXT There is an abnormal increase in TGF-β1 bioavailability in women with polycystic ovary syndrome (PCOS), which might play a role in the pathophysiology of this syndrome. Vitamin D (VD) supplementation improves various clinical manifestations of PCOS and decreases TGF-β1 levels in several diseases including myelofibrosis. OBJECTIVE The objective of the study was to determine the effect of VD supplementation on TGF-β1 bioavailability in VD-deficient women with PCOS and assess whether changes in TGF-β1/soluble endoglin (sENG) levels correlate with an improvement in PCOS clinical manifestations. DESIGN This was a prospective, randomized, placebo-controlled trial. SETTING The study was conducted at an academic-affiliated medical center. PARTICIPANTS Sixty-eight VD-deficient women with PCOS who were not pregnant or taking any exogenous hormones were recruited between October 2013 and January 2015. INTERVENTIONS Forty-five women received 50 000 IU of oral vitamin D3 and 23 women received oral placebo once weekly for 8 weeks. MAIN OUTCOMES MEASURES Serum TGF-β1, sENG, lipid profile, testosterone, dehydroepiandrosterone sulfate, and insulin resistance were measured. The clinical parameters were evaluated before and 2 months after treatment. RESULTS The VD level significantly increased and normalized after VD supplementation (16.3 ± 0.9 [SEM] to 43.2 ± 2.4 ng/mL; P < .01), whereas it did not significantly change after placebo. After the VD supplementation, there was a significant decrease in the following: the interval between menstrual periods (80 ± 9 to 60 ± 6 d; P = .04), Ferriman-Gallwey score (9.8 ± 1.5 to 8.1 ± 1.5; P < .01), triglycerides (138 ± 22 to 117 ± 20 mg/dL; P = .03), and TGF-β1 to sENG ratio (6.7 ± 0.4 to 5.9 ± 0.4; P = .04). In addition, the ΔTGF-β1 to sENG ratio was positively correlated with Δtriglycerides (r = 0.59; P = .03). CONCLUSIONS VD supplementation in VD-deficient women with PCOS significantly decreases the bioavailability of TGF-β1, which correlates with an improvement in some abnormal clinical parameters associated with PCOS. This is a novel mechanism that could explain the beneficial effects of VD supplementation in women with PCOS. These findings may support new treatment modalities for PCOS, such as the development of anti-TGF-β drugs.

Transforming growth factor-β1 and its receptor soluble endoglin are altered in polycystic ovary syndrome during controlled ovarian stimulation.

OBJECTIVE To evaluate the relationship between transforming growth factor (TGF)-β1 and its receptor, soluble endoglin (sENG), in the serum and follicular fluid of women with polycystic ovarian syndrome (PCOS) compared with that of non-PCOS normal ovulating women during controlled ovarian stimulation (COS). DESIGN Prospective case-control study. SETTING Academic-affiliated assisted reproductive technology unit. PATIENT(S) Fourteen PCOS and 14 matched non-PCOS control women undergoing COS. INTERVENTION(S) Serum was collected on day 3 (baseline), day of hCG, and day of retrieval. Follicular fluid (FF) was collected on day of oocyte retrieval. ELISA was performed to determine TGF-β1 and sENG protein levels. MAIN OUTCOME MEASURE(S) Serum and FF levels of TGF-β1 and sENG. RESULT(S) Serum TGF-β1 did not change significantly during COS but was increased in PCOS compared with non-PCOS women on day 3 and days of hCG administration and oocyte retrieval. Serum sENG increased after hCG administration only in the non-PCOS control group. In addition, serum sENG was decreased in PCOS compared with non-PCOS control women on the days of hCG and retrieval. Accordingly, the bioavailability of TGF-β1 (TGF-β1/sENG ratio) was increased in women with PCOS compared with non-PCOS controls at all three time points. No differences in either factor were noted in FF between groups. CONCLUSION(S) The increased TGF-β1 bioavailability in PCOS is not only due to increased TGF-β1 levels but also to decreased levels of its receptor, sENG. These data suggest that increased TGF-β1 bioavailability may contribute to the pathogenesis of PCOS and its increased risk for ovarian hyperstimulation.

Follicular fluid placental growth factor is increased in polycystic ovarian syndrome: correlation with ovarian stimulation

BackgroundPolycystic ovarian syndrome (PCOS) is characterized by increased ovarian angiogenesis and vascularity. Accumulating evidence indicates that vascular endothelial growth factor (VEGF) is increased in PCOS and may play an important role in these vascular changes and the pathogenesis of this disease. Placental growth factor (PlGF), a VEGF family member, has not been previously characterized in PCOS women. We investigated levels and temporal expression patterns of PlGF and its soluble receptor sFlt-1 (soluble Fms-like tyrosine kinase) in serum and follicular fluid (FF) of women with PCOS during controlled ovarian stimulation.MethodsThis was a prospective cohort study of 14 PCOS women (Rotterdam criteria) and 14 matched controls undergoing controlled ovarian stimulation. Serum was collected on day 3, day of hCG and day of oocyte retrieval. FF was collected on retrieval day. PlGF, sFlt-1 and anti-mullerian hormone (AMH) protein concentrations were measured using ELISA. Since sFlt-1 binds free PlGF, preventing its signal transduction, we calculated PlGF bioavailability as PlGF/sFlt-1 ratio.ResultsSerum PlGF and sFlt-1 levels were constant throughout controlled ovarian stimulation, and no significant differences were observed in either factor in PCOS women compared with non-PCOS controls at all three measured time points. However, FF PlGF levels were increased 1.5-fold in PCOS women compared with controls (p < 0.01). Moreover, FF PlGF correlated positively with number of oocytes retrieved and the ovarian reserve marker anti-mullerian hormone (AMH) and negatively with age. In addition, FF sFlt-1 levels were decreased 1.4-fold in PCOS women compared to controls (p = 0.04). PlGF bioavailability in FF was significantly greater (2-fold) in PCOS women compared with non-PCOS controls (p < 0.01).ConclusionsThese data provide evidence that FF PlGF correlates with ovarian stimulation and that its bioavailability is increased in women with PCOS undergoing controlled ovarian stimulation. This suggests that PlGF may play a role in PCOS pathogenesis and its angiogenic dysregulation.

Angiopoietin-1 and angiopoietin-2 are altered in polycystic ovarian syndrome (PCOS) during controlled ovarian stimulation

Polycystic ovarian syndrome (PCOS) ovaries are characterized by increased angiogenesis and hypervascularity. While angiopoietin-1 (Ang-1) and its antagonist, angiopoietin-2 (Ang-2), are essential for ovarian function and angiogenesis, the levels of Ang-1 and Ang-2 in PCOS are unknown. This was a prospective cohort study of 14 PCOS women and 14 matched controls undergoing controlled ovarian stimulation (COS). Serum was collected on day 3, hCG and retrieval days. Follicular fluid (FF) was collected on retrieval day. Serum Ang-1 and Ang-2 levels were constant throughout COS, but serum Ang-1 levels were increased at all time points in PCOS women compared with controls (p < 0.05). No differences between groups were found in serum Ang-2 levels or FF Ang-1 levels. However, FF Ang-2 levels were increased almost 2-fold in PCOS women compared with controls (p < 0.01), and correlated positively with number of oocytes retrieved (r = 0.65, p < 0.0001). This study is the first to provide evidence of an alteration in the Ang-1/Ang-2 system in PCOS women. The biological role of Ang-2 in promoting capillary leakage, the increased Ang-2 FF level in PCOS, and its correlation with number of oocytes suggest that Ang-2 may play an important role in the increased risk of ovarian hyperstimulation in PCOS.

Vitamin D decreases serum VEGF correlating with clinical improvement in vitamin D-deficient women with PCOS: A randomized placebo-controlled trial

Vascular endothelial growth factor (VEGF) has been suggested to play a role in the pathophysiology of polycystic ovary syndrome (PCOS) and may contribute to increased risk of ovarian hyperstimulation syndrome (OHSS) in affected individuals. Vitamin D (VitD) supplementation improves multiple clinical parameters in VitD-deficient women with PCOS and decreases VEGF levels in several other pathologic conditions. Unveiling the basic mechanisms underlying the beneficial effects of vitamin D on PCOS may enhance our understanding of the pathophysiology of this syndrome. It may also suggest a new treatment for PCOS that can improve it through the same mechanism as vitamin D and can be given regardless of vitamin D levels. Therefore, we aimed to explore the effect of VitD supplementation on serum VEGF levels and assess whether changes in VEGF correlate with an improvement in characteristic clinical abnormalities of PCOS. This is a randomized placebo-controlled trial conducted between October 2013 and March 2015. Sixty-eight VitD-deficient women with PCOS were recruited. Women received either 50,000 IU of oral VitD3 or placebo once weekly for 8 weeks. There was a significant decrease in serum VEGF levels (1106.4 ± 36.5 to 965.3 ± 42.7 pg·mL–1; p < 0.001) in the VitD group. Previously reported findings of this trial demonstrated a significant decrease in the intermenstrual intervals, Ferriman-Gallwey hirsutism score, and triglycerides following VitD supplementation. Interestingly, ∆VEGF was positively correlated with ∆triglycerides (R2 = 0.22; p = 0.02) following VitD supplementation. In conclusion, VitD replacement significantly decreases serum VEGF levels correlating with a decrease in triglycerides in women with PCOS. This is a novel molecular explanation for the beneficial effects of VitD treatment. It also suggests the need to investigate a potential role of VitD treatment in reducing the incidence or severity of OHSS in VitD-deficient women with PCOS.

Anti-Müllerian hormone as an independent predictor of twin versus singleton pregnancy in fresh cycles.

This study evaluated anti-Müllerian hormone (AMH) as a possible predictor of twin pregnancy in women undergoing fresh cycles who had more than one embryo transferred. A retrospective study was performed of 139 patients undergoing fresh non-donor cycles which resulted in either singleton or twin pregnancy between 2009 and 2010 in this fertility clinic. Random serum AMH and other clinically relevant variables were compared. For further analysis, the population was stratified by age (<34 and ⩾34 years). Random serum AMH concentrations were 1.4-times greater in women conceiving twins compared with singletons (P=0.03). In women aged ⩾34, the AMH concentration in twins was 1.8-fold greater than singletons (P=0.001). Multivariate analysis demonstrated that AMH was an independent predictor of twins. ROC curve analysis showed that AMH had a significant predictive ability for twin pregnancy in women aged ⩾34 (AUC 0.67, P=0.01). In contrast, in women aged <34, AMH was not different between twin and singleton pregnancies. In summary, random serum AMH is an independent predictor of twin gestation when more than one embryo is transferred in women aged ⩾34. Considering a womans AMH before transferring more than one embryo may assist in reducing the incidence of twins. Transferring multiple embryos to increase the chance of pregnancy and live birth rate is a common practice among assisted reproduction providers but often results in a high proportion of multiple pregnancies. Identifying factors predictive of multiple pregnancies is of paramount importance for the successful development of strategies to minimize multiple gestations. Anti-Müllerian hormone (AMH) has been shown to be closely correlated with a womans egg reserve. Aside from the strong association of serum AMH concentration with quantitative ovarian response, serum AMH concentration has been associated with qualitative aspects of assisted reproduction such as embryo quality and pregnancy rates. Our objective was to evaluate AMH as a possible predictor of twin pregnancy in women undergoing fresh cycles who had more than one embryo transferred. Our study included 139 patients undergoing fresh non-donor cycles which resulted in either a singleton or twin pregnancy between 2009 and 2010 in our fertility clinic. Random serum AMH concentrations were compared. For further analysis, the population was divided into two groups (age <34 and ⩾34). AMH concentrations were 1.4-times greater in women conceiving twins compared with singletons. In women aged ⩾34, the AMH concentration in twins was 1.8-fold greater than singleton pregnancies. AMH was an independent predictor of twins with a significant predictive ability for twin pregnancy in women aged ⩾34. In contrast, in women aged <34, AMH was not different between twin and singleton pregnancies. In conclusion, considering a womans AMH before transferring more than one embryo may assist in reducing the incidence of twins.