Low Antral Follicle Count Research Articles

Expression of genes involved in BMP and estrogen signaling and AMPK production can be important factors affecting total number of antral follicles in ewes

Follicular growth and ovulation of healthy oocytes is a complicated process which is regulated by several endocrine and paracrine factors as well as cross-talk between the oocyte and its surrounding somatic cells. This study compared the expression profile of some candidate genes involved in BMP signaling as well as estrogen and AMPK production in cumulus-oocyte complex (COC) of small and large antral follicles and their associated somatic cell layers in ovaries from ewes with high- and low-antral follicle count (AFC). Expression of GDF9 was increased by increasing the size of antral follicles, while BMP15 expression was decreased by follicular size. It should be noteworthy that transcription of both GDF9 and BMP15 was also detected in the adjacent cellular layers under the follicles. There was a very strong positive correlation between BMP15 and BMPR2 in ovary tissues. Expression of GDF9 was highly correlated with BMP15, BMPR1B, and BMPR2 in large antral follicles. Expression of BMP7 in small antral follicles and BMPR2 in ovary tissues was significantly increased in the high-AFC group. Expression of ESR1 and ESR2 involved in estrogen production as well as PRKAA1 which involved in AMPK production were significantly greater in large antral follicles of high-AFC. There was a very high correlation between Cyp19 and ESR1 in large antral follicles and ovary tissues. Expression of Cyp19 and PRKAA1 were positively correlated with GDF9, BMP15, and BMP7 in large follicles. In conclusion, this study suggests that apart from the BMP signaling, genes involved in AMPK and estrogen production can be pivotal players in ewe's follicular development process. In addition, a strong cross-talk can exist among AMPK, BMP signaling, and estrogen synthesis systems in ewe ovary.

Live birth rates are satisfactory following multiple IVF treatment cycles in poor prognosis patients

This seven-year retrospective study analysed the live birth rate (LBR) for women undergoing IVF treatment with various antral follicle counts (AFC). The LBR decreased with lower AFC ratings, and in 290 treatment cycles for women in the poorest AFC category, ≤4 follicles (group E), the LBR was the lowest at 10.7%. The pregnancy loss rate (PLR) significantly increased with poorer AFC categories, from 21.8% in AFC group A (≥20 follicles), to 54.4% in AFC group E (p<0.0001). This trend was repeated with advancing age, from 21.6% for younger women (<35years), to 32.9, 48.5 and 100% for ages 35–39, 40–44 and ≥45 years, respectively (p<0.0001). However, LBR within the specific AFC group E cohort was also age-dependent and decreased significantly from 30.0% for <35 years old, to 13.3, 3.9 and 0% for patients aged 35–39, 40–44 and ≥45 years, respectively. Most, importantly, LBR rates within these age groups were not dependent on the number of IVF attempts (1st, 2nd, 3rd or ≥4 cycles), which indicated that cycle number should not be the primary deciding factor for cessation of IVF treatment in responding women <45years old.

144 EFFECT OF ANTRAL FOLLICLE COUNT IN BEEF HEIFERS ON IN VITRO FERTILIZATION/PRODUCTION

Our objective has been to compare the IVF and in vitro production (IVP) of embryos from low and high antral follicle count (AFC) heifers. This is the fourth year of the study with years 1 to 3 reported individually. For this report, we add data for the fourth year and present a combined analysis (years 1 to 4) for the first time. Each year, AFC was determined on ~120 Angus heifers using transrectal ultrasonography. Ten heifers with the lowest AFC and 10 heifers with the highest AFC and all with evidence of oestrous cyclicity were synchronized with two 5-mL injections of PGF2α 11 days apart. Half were harvested on Day 5 to 6 and half on Day 15 to 16 of the oestrous cycle. The IVF procedure was slightly modified each year. For year 4, the IVF procedure included protocols for semi-defined media and was as described (IVP Protocol, P. J. Hansen’s Laboratory, University of Florida). Cumulus-oocyte complexes (COC) from follicles less than 8 mm in diameter were cultured in maturation medium (5% CO2; 38.5°C) for 24 h. Matured COC were fertilized using thawed frozen semen from a bull that was purified using isolate. Motile spermatozoa were added to COC in fertilization medium at a final concentration of 1 × 106 spermatozoa per mL. About 24 h later, presumptive zygotes were placed in micro drops of development medium under oil, and cultured (5% CO2; 5% O2; balance N2; 38.5°C). On Day 3 and 8 after fertilization, cleavage and blastocyst development rates, respectively, were assessed. Data were analysed using the MIXED procedure of SAS (SAS Institute Inc., Cary, NC, USA) and the model included the effects of year (1 to 4), group (high or low AFC), and their interaction. The year × group interaction was not significant (P &gt; 0.10). Low AFC heifers, compared with high AFC heifers, had fewer numbers of COC (P &lt; 0.0001; 12.8 ± 1.83 v. 31.9 ± 1.86), fewer numbers of COC that cleaved (P &lt; 0.0001; 8.0 ± 1.38 v. 21.6 ± 1.40), and fewer numbers of COC that developed to the blastocyst stage (P &lt; 0.0001; 1.7 ± 0.58 v. 5.7 ± 0.58). Year affected the numbers of COC that cleaved (P &lt; 0.003) and the numbers of COC that developed to the blastocyst stage (P &lt; 0.0001). Year also influenced the percentage of COC that cleaved (P &lt; 0.0002) and the percentage of COC that developed to blastocysts (P &lt; 0.0001). Group (AFC) did not influence (P &gt; 0.19) the percentage of COC that cleaved (61.2 ± 2.83 v. 66.4 ± 2.83%, for low v. high AFC, respectively). Low AFC heifers had a lower (P &lt; 0.002) percentage of COC that developed to blastocysts (10.3 ± 1.52%) than high AFC heifers (17.6 ± 1.52%). These results indicate that high AFC heifers, compared to low AFC heifers, have more COC recovered, more COC cleaved, and more COC developed to the blastocyst stage. The percentage of COC cleaved did not differ between AFC groups; however, the percentage of COC that developed to the blastocyst stage was greater for high than low AFC heifers. This suggests a potential advantage in maternal to embryonic transition for high compared with low AFC heifers.

New automated AntiMüllerian hormone (AMH) assays are more reliable than the manual assay in patients with reduced antral follicle count (AFC)

To improve quality of AMH measurements, fully automated AMH assays have been recently developed. Yet, analytical data available failed to give emphasis on the necessary relationship between AMH and the number of AMH-producing antral follicles as a quality reference. Moreover, the relative performance of new automated and manual AMH assays in subgroups of patients displaying low, intermediate, and high AFC remained undetermined. Therefore, we decided to investigate the reliability and concordance of AMH levels obtained by one manual and two automated AMH assays in different AFC populations. Prospective study. Serum AMH levels were measured in frozen-thawed serum aliquots from 211 women, aged 24 to 43 years, using one manual (AMH Gen II) and two fully automated (Access AMH and Elecsys AMH) assays. AFC was performed on cycle-days 1-5 (same day of bloodwork) under strictly standardized conditions and sorted into 3 groups (tercile values): low AFC (3-12 follicles; n=73), intermediate AFC (13-20 follicles; n=65), and high AFC (21-84 follicles; n=73). The strenght of relationships between AFC and AMH levels was assessed by the Spearman correlation test and Spearman correlation coefficients were compared using the Fisher r-to-z transformation. Concordance among AMH assays was assessed graphically using the Bland-Altman plots and by Passing-Bablok regression. Overall, circulating AMH levels were lower (P< 0.001) with Access AMH (-16%) and Elecsys AMH (-20%) than with AMH Gen II. The limited amount of AMH values situated outside the ± 2 standard deviation interval in Bland-Altman plots (4.7% between AMH Gen II and Access AMH; 4.3% between AMH Gen II and Elecsys AMH; and 2.8% between Access and Elecsys AMH) indicated adequate concordance among assays, a phenomenon that tended to be improved in the low AFC group. Further, AFC was strongly correlated with AMH levels obtained with the 3 assays (r=0.83, P<0.001; r=0.83, P<0.001; and r=0.83, P<0.001, respectively) thereby confirming their comparable reliability. It is noteworthy that, in the low AFC group, AMH levels obtained by Access AMH and Elecsys AMH showed a stronger correlation with AFC (r=0.63; P<0.03 and r=0.65; P<0.005, respectively) than the AMH Gen II assay (r=0.52). This improved performance was not observed in the intermediate and the high AFC groups. As compared to conventional AMH Gen II assay results: 1. Serum AMH concentrations were -16% and -20% lower with Access AMH and Elecsys AMH, respectively, but concordance among techniques was adequate; 2. It was remarkable that both automated AMH assays were significantly more strongly correlated with AFC than the manual assay in patients endowed with few antral follicles. This constitutes a considerable advantage of the new AMH assays because this subset of patients is the main target for accurate ovarian function assessments.

In women undergoing elective egg cryopreservation with less-than-expected ovarian reserve a 4 month break from combined hormonal contraceptives is associated with a 50% increase in oocyte yield

Unlike the infertility setting, many women who present for elective oocyte freezing cycles are taking long-term (>1 year) combined hormonal contraceptives (OCP). Based on the hypothesis that long-term OCP use may cause ovarian suppression, we routinely recommend a break from OCP use prior to ovarian stimulation in women with low Antral Follicle Count (AFC).

Effect of salpingectomy, ovarian cystectomy and unilateral salpingo-oopherectomy on ovarian reserve

Pelvic surgery can affect ovarian reserve, but estimates of the potential effect of different surgical procedures are lacking. This study examines the markers of ovarian reserve after different procedures in order to help the provision of informed consent before surgery. Anti-Müllerian hormone (AMH), antral follicle count (AFC) and follicle-stimulating hormone (FSH) of women with a history of salpingectomy, ovarian cystectomy or unilateral salpingo-oophorectomy were compared to those without history of surgery using cross-sectional data adjusting for patient and clinical factors in multivariable regression model. There were 138 women who had had salpingectomy, 36 unilateral salpingo-oopherectomy, 41 cystectomy for ovarian cysts that are other than endometrioma and 40 women had had excision of endometrioma. There was no significant difference in AMH (9 %; p = 0.33), AFC (−2 %; p = 0.59) or FSH (−14 %; p = 0.21) in women with a history of salpingectomy compared to women without surgery. Women with a history of unilateral salpingo-oophorectomy were found to have significantly lower AMH (−54 %; p = 0.001). These women also had lower AFC (−28 %; p = 0.34) and higher FSH (14 %; p = 0.06), the effect of which did not reach statistical significance. The study did not find any significant associations between a history of cystectomy, for disease other than endometrioma and AMH (7 %; p = 0.62), AFC (13 %; p = 0.18) or FSH. (11 %; p = 0.16). Women with a history of cystectomy for ovarian endometrioma had 66 % lower AMH (p = 0.002). Surgery for endometrioma did not significantly affect AFC (14 %; p = 0.22) or FSH (10 %; p = 0.28). Salpingo-oopherectomy and cystectomy for endometrioma cause a significant reduction in AMH levels. Neither salpingectomy nor cystectomy for cysts other than endometrioma has appreciable effects on ovarian reserve.Electronic supplementary materialThe online version of this article (doi:10.1007/s10397-016-0940-x) contains supplementary material, which is available to authorized users.

The relationship between antral follicle count in a bovine ovary and developmental competence of in vitro-grown oocytes derived from early antral follicles.

To clarify the relationship between ovarian reserve and the developmental competence of bovine oocytes, oocyte-granulosa complexes (OGCs) collected from early antral follicles (≤ 1 mm) in ovaries with high (≥ 25) and low (<25) antral follicle counts (AFCs) were used. OGCs derived from different AFC groups were cultured for growth followed by maturation, fertilization and blastocyst formation. Viability of OGCs during growth culture was similar between groups; however, OGCs in the high-AFC group had a larger number of granulosa cells than the low-AFC group at 12 days of growth. The proportion of matured oocytes in the high-AFC group was higher than that in the low-AFC group. Mitochondrial activity of oocytes before maturation in the high-AFC group was higher than that in the low-AFC group; however, accumulation of reactive oxygen species was similar between groups. Cleavage rate in the high-AFC group tended to be higher than that in the low-AFC group, although blastocyst development was similar between groups. In conclusion, oocytes derived from ovaries with high AFC have higher maturational ability and fertilizability than those from low AFC. The difference may be caused by high proliferation of granulosa cells from ovaries with high AFC.

Assessment of ovarian reserve: Anti-Mullerian hormone versus follicle stimulating hormone.

Background:This study aimed to evaluate the strength of anti-Mullerian hormone (AMH) and follicle stimulating hormone (FSH) in reflecting the antral follicle count (AFC) in infertile females.Materials and Methods:This cross-sectional study was conducted on 160 females, visiting infertility clinic for assisted reproduction. Serum samples collected on the 3rd day of the cycle were assayed for FSH, luteinizing hormone, and AMH while AFC was assessed via transvaginal ultrasound. The study cohort was segregated into three groups based on AFC.Results:Chronological age and FSH was significantly high in females with very low AFC (P < 0.01 and 0.009, respectively), yet they failed to discriminate patients with normal and higher follicle count (P = 0.65 and 0.84). Conversely, AMH reported highly significant difference between very low AFC and with those having either normal AFC (P = 0.002) or higher AFC (P = 0.001). Moreover, a significant difference in AMH was observed between normal and higher AFC group (P = 0.04).Conclusion:Compared to female’s age and FSH, AMH is superior in clustering study cohort on the bases of antral follicular pool, especially in setups with nonavailability of technological expertise to assess AFC. Incorporation of AMH along with other biomarkers improves estimation of baseline ovarian reserve, required to standardize dose for optimum response; avoiding the risk of failure to retrieve oocyte or inappropriate stimulation leading to ovarian hyperstimulation syndrome. Further prospective studies are required to ascertain its role in predicting the outcomes of ART in such patients.

Antral follicle count: Is the right ovary more predictive than the left for live birth?

Objective: The objective of this study was to assess the impact of discordant ovarian antral follicle counts (AFCs) on cycle stimulation and live birth in autologous in vitro fertilization (IVF) cycles. Materials and Methods: This is a retrospective analysis of first-time cycles of 153 patients undergoing gonadotropin-releasing hormone-antagonist IVF. Results: While AFC significantly correlated with cycle stimulation characteristics, only the right ovarian AFC significantly correlated with live birth (r = 0.18, P< 0.02). Right ovarian AFC was significantly greater in live birth cycles (12.3 ± 7.8) compared to cycles without a live birth (9.6 ± 6.0, P< 0.02). Using ≤ 7 as a threshold of low unilateral AFC, concordant low AFC was present in 28% (Group 1: n = 43); discordant low left and normal right in 13% (Group 2: n = 19); discordant low right and normal left in 11% (Group 3: n = 17); and concordant normal in both ovaries in 48% (Group 4: n = 74) of patients. Live birth was similar in Group 2 (36.8%), Group 3 (35.3%), and Group 4 (37.8%), but significantly less in Group 1 (9.3%) (P < 0.05). Conclusion: Live birth was higher with greater right AFC. Using threshold AFC, adverse outcomes were only noted when both ovaries had low counts. Individual AFC may serve as a more specific indicator than total AFC as a marker of ovarian reserve.

181 IN VITRO FERTILIZATION (IVF) USING SEMI-DEFINED CULTURE CONDITIONS FROM LOW OR HIGH ANTRAL FOLLICLE COUNT PUBERTAL BEEF HEIFERS

To compare the in vitro fertilization (IVF) and production (IVP) of embryos from low and high antral follicle count (AFC) heifers, AFC were determined on 106 heifers using transrectal ultrasonography. Ten heifers with the lowest AFC (avg. 13.2) and 10 heifers with the highest AFC (avg. 27.4) with evidence of oestrous cyclicity were synchronised with 2 injections of prostaglandin F2α (PGF2α); half were harvested on Days 5 to 6 and half on Days 15 to 16 of the oestrous cycle. The IVF procedures included protocols for semi-defined media and were as described (IVP Protocol, P. J. Hansen’s Laboratory, University of Florida, Gainesville, FL, USA). Cumulus-oocyte complexes (COC) from follicles less than 8 mm in diameter were cultured in maturation medium (5% CO2; 38.5°C) for 24 h. Matured COC were fertilised using thawed frozen semen from a crossbred bull that was purified using Percoll gradient separation procedures. Motile spermatozoa were added to COC in fertilization medium at a final concentration of 1 × 106 spermatozoa per mL. About 24 h later, presumptive zygotes were placed in microdrops of development medium under oil, and cultured (5% CO2, 5% O2, balance N2, 38.5°C). On Days 3 and 8 after fertilization, cleavage and blastocyst development rates, respectively, were assessed. Data were analysed using the mixed procedure of SAS (SAS Institute, Cary, NC, USA) and the model included the effects of collection day, group (high or low AFC), and their interaction. More COC (P &lt; 0.0005) were collected from high than low AFC heifers (30.3 v. 9.3 ± 3.12 per heifer). Both the number and percentage of COC that cleaved had an interaction between collection day and group (P &lt; 0.03). The interaction appeared to be due to low cleavage and development rates on 1 of 4 collection days (appeared not related to day of oestrous cycle). Although high compared to low AFC heifers had more COC that cleaved (18.7 v. 4.4 ± 1.84 per heifer), the percentage of cleaved COC did not differ (59.2 v. 49.8 ± 3.36%). There were no significant differences between high and low AFC heifers in the number of blastocysts (3.1 v. 0.6 ± 1.21 per heifer) or in the percentage of COC that developed to blastocysts (8.8 v. 5.2 ± 3.60%). In previous replicates (years), we reported that high AFC heifers had more COC collected, more COC that cleaved, and more COC that developed to blastocysts than low AFC heifers. In contrast, in this study numbers of COC that developed to blastocysts did not significantly differ between high and low AFC heifers. Additionally, the percentage of COC that cleaved, and that developed to blastocyst have been similar between high and low AFC heifers. Therefore, high compared to low AFC heifers may produce more IVP embryos; however, AFC does not appear to affect the competence of an oocyte to develop and mature to the blastocyst stage.

130 ANTRAL FOLLICLE COUNTS, VULVA WIDTH, AND SERUM ANTI-MULLERIAN HORMONE CONCENTRATIONS IN BOVINE FEMALES OF THE TABAPUÃ BREED

Bovine genetic selection for fertility is difficult in part due to the low heritability of fertility traits and to the extension of the calving interval and the productive life length inherent to this species, which makes it harder to identify superior individuals from their records. In this regard, it is feasible to try to identify characteristics that are easy to detect and may be linked to superior reproductive efficiency as early as possible in females. The objective was to evaluate the possible relationships between vulva width, antral follicle counts (AFC), and the serum concentrations of anti-Mullerian hormone (AMH). Tabapuã (a Brazilian Zebu breed) heifers (n = 155) and cows (n = 422) from 4 herds were used in the study in which all antral follicles ≥3 mm were counted directly with an ultrasound unit. Follicle counts were classified into 2 classes, based on the methodology proposed by Burns et al. (2005) in dairy cattle, such that the low and intermediate AFC classes were merged into one class (low; n = 245) and the high and very high into another (high; n = 332). The vulva width was determined by measuring the distance between the lateral borders of the vulva with a digital caliper placed at a 90-degree angle from the half point of the rima vulvae. Single blood samples were collected from a subgroup of animals (n = 220) and the serum frozen (–20°C) after centrifugation for the AMH analysis, which was performed by specific bovine ELISA (Ansh Laboratories, Webster, TX, USA) test. All statistical analyses were performed with SAS (SAS Institute Inc., Cary, NC, USA). Data was submitted for analyses of variance using the PROCGENMOD procedure. Vulva width was larger (P = 0.02) in the class of animals of the high AFC (8.73 ± 0.11 cm) in comparison with those of the low AFC (8.39 ± 0.09 cm), considering the larger sample (n = 591). In the subsample (n = 220) the concentration of AMH was higher (P &lt; 0.0001) in the high AFC class females (1.15 ± 0.07 ng mL–1; n = 95) in comparison with the low AFC class (0.50 ± 0.06 ng mL–1; n = 125). Vulva width, AFC, and AMH serum concentrations were interrelated and likely fertility indicative phenotypes under the present experimental conditions and should be further evaluated in larger samples in order to be considered in bovine genetic selection.

Postnatal pituitary and follicular activation: a revisited hypothesis in a sheep model.

The importance of postnatal pituitary activation as regards female reproductive development is not yet understood. By taking advantage of the experimental model developed in a previous study, i.e. ewe lambs expressing markedly different ovarian phenotypes at 50 days of age, we designed this study to determine whether differences found in ovarian status during the early prepubertal period are due to different patterns of postnatal pituitary activation, and to assess whether these differences have long lasting effects on subsequent reproductive performance. Results showed that ewe lambs with high antral follicle count (AFC) at 50 days of age had significantly lower plasma FSH concentrations and higher anti-Mullerian hormone (AMH) concentrations during the first 9 weeks of age compared with low AFC ewe lambs (P<0.0001). With a longitudinal experiment we showed that a high AFC in the early prepubertal period is associated with consistently higher AMH concentrations and numbers of antral follicles up to the postpubertal period, and with higher pregnancy rates in the first breeding season. In addition, the effect of age in decreasing AMH concentrations was more marked in the low AFC group. Results of the present study demonstrate that ewe lambs undergo different patterns of postnatal pituitary activation. A high AFC at 50 days of age indicates an advanced phase of ovarian maturation, which was accompanied by constantly higher AMH concentrations up to the postpubertal period, a greater ovarian response to FSH stimulation and by higher pregnancy rates at first mating, as compared with the low AFC group.

Fertilizability of oocytes derived from Holstein cows having different antral follicle counts in ovaries

In this study, to clarify the relationship between ovarian reserve and oocyte quality, cumulus-oocyte complexes (COCs) were collected repeatedly by ovum pick-up (OPU) from cows with high and low antral follicle counts (AFCs) at short (3–4 days) and long (7 days) intervals, and COC morphologies and oocyte fertilizability were examined. The relationship between AFC and follicular growth after OPU was also investigated. Cows showing AFC of ≥30 in at least one OPU session were grouped into the high-AFC group. At a short interval, follicular sizes and COC morphologies were similar between the different AFC groups. However, the normal fertilization rate was higher in the high-AFC group than in the low one, although total penetration rates were similar. At a long interval, the percentage of COCs with poor morphology in the high-AFC group was higher and the normal fertilization rate was lower than in the low one. In the low-AFC group, normal fertilization rates at short and long intervals were similar, and mean follicular size became larger at a long than at a short interval. However, mean follicular sizes at short- and long-interval OPU were similar in the high-AFC group. In conclusion, it is suggested that oocytes derived from cows with high AFC had higher fertilizability than those from cows with low AFC when OPUs were performed at a short (3–4 days) interval. However, oocyte quality in high-AFC cows was impaired by long-interval (7 days) OPU, possibly due to the degradation of follicles.

Association between antral follicle count and reproductive measures in New Zealand lactating dairy cows maintained in a pasture-based production system

The antral follicle count (AFC) in cattle is consistent throughout the estrous cycle of individual cows, and cows with a lower AFC have lower fertility. We assessed the AFC at random stages of the estrous cycle, examined the correlation between AFC classifications, and determined the relationship between the most rapid and practical laboratory-based AFC classification (AFC of follicles of ≥2 mm in diameter) and fertility measures in New Zealand lactating dairy cows. Cows detected in estrus (n = 202) or not (n = 239) during the first 4 weeks of the breeding season were subjected to ultrasonography and classified as having a high, medium, or low AFC at the time of scanning (on-site classification). Images from ultrasound scanning were recorded onto video for accurate follicle counting in an imaging laboratory. A strong association (P < 0.05) between the AFC of follicles with a diameter of 2 mm or greater and fertility was observed. Cows with a high AFC had a shorter (P < 0.05) interval from calving to conception by artificial insemination (AI; 82.4 ± 1.6 vs. 87.3 ± 1.2 days) and greater pregnancy rates (PRs; i.e., PR to the first AI [68.1% vs. 45.3%], 6-week PR [81.9% vs. 67.3%], and overall PR [91.3% vs. 79.7%]) than cows with a low AFC. The AFC was positively associated (P < 0.0001) with age. Progesterone concentrations during diestrus were greater (P < 0.05) in high-AFC cows (7.6 ± 0.3 ng/mL) than in low-AFC cows (6.5 ± 0.3 ng/mL), whether these were pregnant (7.7 ± 0.3 ng/mL) or not (6.3 ± 0.2 ng/mL). A rapid on-site scoring system determined that cows classified as having a high AFC had a shorter (P < 0.05) interval from calving to the first AI (76.5 ± 1.7 vs. 82.3 ± 1.9 days) and were more likely to show estrus (P < 0.01; 56.8% vs. 36.4%) and have a CL at the beginning of the breeding season (P < 0.01; 93.4% vs. 79.6%) than cows with a low on-site AFC. Collectively, we have confirmed an association between AFC2 and fertility, and these results support the hypothesis that cows with a greater number of antral follicles are more fertile than cows with a lesser number of follicles. Although the on-site classification was related to resumption of estrous cycles after calving, associations with other fertility measurements could not be observed, highlighting a need for further refinement of the on-site classification system for rapid phenotyping of the AFC.

Reduced ovarian reserve in patients with adult polymyositis.

Polymyositis (PM) affects female gender during reproductive age; however, there is no study assessing ovarian reserve in these patients to evaluate ovarian reserve markers in PM. Eight female patients with PM (Bohan and Peter criteria, 1975) with aged 18-40years, followed at our tertiary centre from March 2011 to May 2014, were invited to participate. They were age-matched with 16 healthy individuals (control group). All were evaluated at early follicular phase of menstrual cycle. Follicle stimulating hormone (FSH), estradiol, inhibin B, anti-Müllerian hormone (AMH) serum levels (ELISA) and sonographic antral follicle count (AFC) were determined. PM patients and controls had comparable mean age (31.4 ± 6.5 vs. 30.7 ± 6.2years, P = 0.946), ethnicity and socioeconomic class (P > 0.05). PM mean age of onset was 27.3 ± 6.5years and disease duration of 6.5 ± 4.1years. Menstrual cycles were alike in both groups with a similar frequency of age at menarche, gynaecological age, duration and length of menstrual cycle (P > 0.05). The median serum level of AMH was significantly lower in PM compared to controls [0.7(0.3-3.4) vs. 3.1(1.4-4.0), P = 0.021]. AMH levels ≤1ng/mL (50 vs. 6.3%, P = 0.024) and very low AFC (37.5 vs. 6.3%, P = 0.037) were significantly in PM patients versus controls. The other hormones (FSH, inhibin B and estradiol levels) were similar between both groups (P > 0.05). The present study was the first to identify subclinical ovarian dysfunction in PM patients during reproductive ages. Further study is necessary to assess the possible role of PM-related factors that may influence the ovarian function of these patients.

Assessment of ovarian reserve by antral follicle count in ovaries with endometrioma.

To evaluate whether the antral follicle count (AFC) is underestimated in the presence of an endometrioma. This was a retrospective cohort study assessing all women undergoing in-vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) at our clinic between January 2011 and December 2012 who had both ovaries and unilateral endometrioma. The primary outcome of the study was the difference between AFC and the number of oocytes retrieved per ovary. Within the study period 787 women underwent IVF/ICSI at our clinic. Sixty of these women had at least one endometrioma, but 23 were excluded from the analysis as six had only one ovary and 17 had bilateral endometriomas. Therefore a total of 37 women were included in this study and analysis. Compared with the contralateral ovaries, ovaries with an endometrioma were significantly larger in volume (median, 10.3 (interquartile range (IQR), 4.7-18.9) cm(3) vs median, 3.6 (IQR, 2.7-6.5) cm(3); P < 0.001) and presented a significantly lower AFC (median, 3.0 (IQR, 1.0-6.0) vs median, 5.0 (IQR, 2.0-6.5); P = 0.001). However, the median number of oocytes retrieved was similar (P = 0.60) between ovaries with an endometrioma (2.0 (IQR, 0.5-5.0)) and the contralateral ovaries (2.0 (IQR, 0.0-4.0)). Accordingly, the median difference between AFC and number of oocytes retrieved was significantly smaller (P = 0.005) for ovaries with an endometrioma (0.0 (IQR, -1.0 to 1.5) than for those without (2.0 (IQR, 0.0-4.0)). Although the AFC is reduced in ovaries with an endometrioma, the number of oocytes retrieved is similar, suggesting that the AFC is underestimated in such ovaries. We believe that this is a consequence of an impaired ability to detect small follicles in the presence of an endometrioma.

Impact of endometriomas and their removal on ovarian reserve.

The effect of endometrioma on ovulatory function and ovarian reserve was unclear. Conflicting opinions exist regarding effect of endometrioma excision on ovarian reserve. Endometriomas do not seem to affect ovulatory function. Women with endometrioma have lower antral follicle count and serum anti-Müllerian hormone levels than age-matched healthy controls. There is high-quality evidence suggesting a significant decline in serum anti-Müllerian hormone levels following endometrioma excision. However, a similarly significant decline in antral follicle count is not demonstrated. Cauterization seems to be a contributing factor to ovarian damage and suturing the cyst bed could perhaps be a better alternative. It seems prudent to warn patients regarding loss of ovarian reserve following endometrioma excision. Surgeons should cautiously limit the use of cauterization following stripping of endometrioma. Well designed studies comparing effect of various haemostatic measures on ovarian reserve are needed.

Abstract P3-12-05: Reduction of ovarian reserve in young early breast cancer patients: 24 months follow up of a prospective cohort trial

Abstract Background: Sideeffects of adjuvant chemotherapy on ovarian function are inadequately studied by now. Premenopausal women undergoing chemotherapy are at risk for sexual hormone deficiency and impaired fertility. This prospective cohort study searches for predictive parameters of ovarian reserve after chemotherapy. Methods: 51 premenopausal patients (average 38 years; 28-46 years) with primary breast cancer have been included. They all received neo- or adjuvant chemotherapy: anthracycline based "A"(FEC)(n= 18); anthracycline/taxane based "T"( TAC or FEC/Doc or EC/Pac)(n=30) or anthracycline- free "AF"(n=3). After chemotherapy patients received endocrine therapy (ET) for hormone receptor (HR) positive disease (tamoxifen (n=34) or goserelin/tamoxifen (n=7)). For HR negative disease (n=20) no ET was given. Before (visit 1) and 6, 12 and 24 months after initiation of chemotherapy (visit 2-4) age and chemotherapy related changes in hormone (LH, FSH, E2 and Anti-Muellerian hormone (AMH)) levels, antral follicle count and amenorrhea as parameters of endocrine function and fertility were assessed. Also quality of life (QL) and additional impact of parity, BMI and nicotine use on ovarian reserve were evaluated. Results: Antral follicle count is decreasing dramatically after chemotherapy and is not fully recovering after 2 years. Antral follicle count before and 1 year after chemotherapy are significantly correlated (p=0.012). Follicle count and age before chemotherapy are negatively correlated (p=0.004). At visit 4 the type of chemotherapy (A, T, AF) has no influence on antral follicle count or AMH levels. Tamoxifen is correlated with a significant decrease in follicle count (vs. no endocrine therapy) (p=0.039). Smoking is significantly detrimental on antral follicle count after 24 months (p=0.001). AMH levels at visits 2-4 are significantly correlated with levels at the 1 visit and are negatively correlated with age (p=0.032). Tamoxifen therapy has no impact on AMH levels. LH and FSH are increasing between visit 1 and 2 and decreasing at visits 3 and 4 (but stay above basic levels). At visit 4 LH is not and FSH is correlated with visit 1 levels (p=0.005). LH and FSH are correlated with age (p=0.0001) and are not influenced by type of chemotherapy or tamoxifen .E2 levels at visit 4 are not influenced by type of chemotherapy, but significantly decreased (p=0.003) by continued smoking. At visit 4 31/51 patients have stayed amenorrheic, 19 resumed their menstruation. This was not influenced by type of chemotherapy or age. Non- smokers are 13 times more likely to resume their menstruation (vs. smokers). Tamoxifen is correlated with a higher probability of permanent amenorrhea (p=0.058, n.s.). BMI, nicotine abuse, age, type of chemotherapy and tamoxifen have no influence on duration of amenorrhea. QL is significantly lower at visit 2, but recovers to basic values at visits 3 and 4. Conclusions: Our study contributes to a better prediction of ovarian reserve before chemotherapy. We suggest personalized counselling on fertility preserving measures before chemotherapy especially at higher age, with low AMH levels or low antral follicle counts. To stop smoking could enhance chances for ovarian preservation. Citation Format: Christoph Mundhenke, Antonia S Wenners, Jana Grambach, Juliane Koss, Ulrike von Hehn, Walter Jonat, Andreas G Schmutzler. Reduction of ovarian reserve in young early breast cancer patients: 24 months follow up of a prospective cohort trial [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P3-12-05.

Concentration of anti-Müllerian hormone in dairy heifers is positively associated with productive herd life

Reliable biomarkers predictive of productive herd life (time in herd after birth of first calf) have heretofore not been discovered in dairy cattle. However, circulating concentrations of anti-Müllerian hormone (AMH) are positively associated with number of follicles or antral follicle count (AFC), ovarian function, and fertility, and approximately 25% of cows have a relatively low AFC and low AMH concentrations. The present study tested the hypothesis that heifers with the lowest AMH concentrations have suboptimal fertility and are removed from a herd for poor reproductive performance at a greater rate, and therefore have a shorter productive herd life compared with age-matched herdmates with higher AMH. To test this hypothesis, 11- to 15-mo-old Holstein heifers (n=281) were subjected to a single measurement of AMH. All heifers not removed from the herd had the opportunity to complete 2 lactations and start their third lactation after calving. During this time, performance and health parameters for each individual were recorded daily by herd managers. Results showed that the quartile of heifers with the lowest AMH concentration also had, on average, a shorter productive herd life (by 196 d), a reduced survival rate after birth of the first calf, the lowest level of milk production (first lactation), the lowest total percentage of cows pregnant (across all lactations), the highest culling rates (first and second lactations and overall), and the highest culling rate for poor reproduction (first lactation) compared with age-matched herdmates with higher AMH. We concluded that a single determination of AMH concentration in young adult dairy heifers may be a simple diagnostic method to predict herd longevity, and AMH may be a useful phenotypic marker to improve longevity of dairy cows.

Second surgery for recurrent unilateral endometriomas and impact on ovarian reserve: a case-control study

To investigate the impact on ovarian reserve of second laparoscopic surgery for recurrent unilateral endometriomas. Case-control study. University teaching hospital. This study included patients who underwent stripping of endometriomas (diameter ≥4cm) and were followed-up at our institution. Case subjects had second surgery for recurrent unilateral endometriomas (n = 18); control subjects had no recurrence and no second surgery (n = 18). This case-control study was based on a retrospective analysis of a prospectively collected database including patients who underwent surgery for endometriomas at our institution. The primary outcome of the study was to assess the changes in antimüllerian hormone (AMH) levels in each study group and between the two study groups. The secondary outcomes of the study were to assess the changes in basal FSH, antral follicle count (AFC), and ovarian volume in each study group and between the two study groups. In both study groups, primary surgery decreased AMH, increased basal FSH, and decreased the AFC of the operated ovary. Before second surgery, case subjects had AMH, basal FSH, and AFC similar to control subjects. After second surgery, case subjects had lower AMH, higher basal FSH, and lower AFC of the affected ovary than before surgery; the volume of the operated ovary was lower than that of the contralateral ovary. The laparoscopic stripping of recurrent ovarian endometriomas is associated with a high risk of ovarian reserve damage and ovarian failure. NCT02047838.