Outcome of assisted reproduction in women with congenital uterine anomalies: a prospective observational study.

OBJECTIVES: To assess the prevalence of congenital uterine anomalies, including arcuate uterus, and their effect on reproductive outcome in subfertile women undergoing assisted reproduction. METHODS: Consecutive women referred for subfertility between May 2009 and November 2015 who underwent assisted reproduction were included in the study. As part of the initial assessment, each woman underwent three-dimensional transvaginal sonography. Uterine morphology was classified using the modified American Fertility Society (AFS) classification of congenital uterine anomalies proposed by Salim et al. If the external contour of the uterus was uniformly convex or had an indentation of < 10 mm, but there was a cavity indentation, it was defined as arcuate or septate. Arcuate uterus was further defined as the presence of a concave fundal indentation with a central point of indentation at an obtuse angle. Subseptate uterus was defined as the presence of a septum, not extending to the cervix, with the central point of the septum at an acute angle; if the septum extended to the internal cervical os, the uterus was defined as septate. Reproductive outcomes, including live birth, clinical pregnancy and preterm birth, were compared between women with a normal uterus and those with a congenital uterine anomaly. Subgroup analysis by type of uterine morphology and logistic regression analysis adjusted for age, body mass index, levels of anti-Müllerian hormone, antral follicle count and number and day of embryo transfer were performed. RESULTS: A total of 2375 women were included in the study, of whom 1943 (81.8%) had a normal uterus and 432 (18.2%) had a congenital uterine anomaly. The most common anomalies were arcuate (n = 387 (16.3%)) and subseptate (n = 16 (0.7%)) uterus. The rate of live birth was similar between women with a uterine anomaly and those with a normal uterus (35% vs 37%; P = 0.47). The rates of clinical pregnancy, mode of delivery and sex of the newborn were also similar between the two groups. Preterm birth before 37 weeks' gestation was more common in women with uterine anomalies than in controls (22% vs 14%, respectively; P = 0.03). Subgroup analysis by type of anomaly showed no difference in the incidence of live birth and clinical pregnancy for women with an arcuate uterus, but indicated worse pregnancy outcome in women with other major anomalies (P = 0.042 and 0.048, respectively). CONCLUSIONS: Congenital uterine anomalies as a whole, when defined using the modified AFS classification, do not affect clinical pregnancy or live-birth rates in women following assisted reproduction, but do increase the incidence of preterm birth. The presence of uterine abnormalities more severe than arcuate uterus significantly worsens all pregnancy outcomes. Copyright © 2017 ISUOG. Published by John Wiley & Sons Ltd.

Assessing repeatability of 3D Doppler indices obtained by static 3D and STIC power Doppler: a combined in-vivo/in-vitro flow phantom study.

OBJECTIVES: To compare the variability in vascularization flow index (VFI) seen in serial acquisitions obtained using spatiotemporal image correlation (STIC) and using conventional static three-dimensional (3D) power Doppler (PD), for both in-vitro and in-vivo models, and to evaluate whether the curves formed by VFI values obtained from successive 'frames' in a STIC dataset are consistent and resemble the waveforms obtained by spectral Doppler analysis. METHODS: The study was divided into two parts: in the first part (the in-vitro model) we scanned a flow phantom, while in the second part (the in-vivo model) we scanned a common carotid artery. Conventional static 3D and STIC-PD datasets were alternately acquired from these two models. VFI values were assessed from 0.38-cm(3) spherical samples of the main flow region in the static 3D datasets and in every frame of the STIC datasets. The variance of the minimum, mean and maximum VFI values from each STIC dataset was compared with the variance of VFI values from the static 3D datasets. RESULTS: Ten static 3D and 10 STIC datasets were acquired from each model. Analysis of the in-vitro and in-vivo models showed a significant reduction in the variance of VFI values obtained using STIC as compared to static datasets. Additionally, we observed that the curves formed by VFI values obtained from successive frames in each STIC dataset were consistent across different datasets and that they resembled the waveforms obtained by spectral Doppler in both models. CONCLUSIONS: 3D-PD indices derived from STIC are more stable than those obtained from conventional static 3D-PD datasets. The curves of VFI throughout a reconstructed cardiac cycle using STIC are repeatable and resemble those obtained by spectral Doppler analysis of the vessel.

Intracycle variation in number of antral follicles stratified by size and in endocrine markers of ovarian reserve in women with normal ovulatory menstrual cycles.

OBJECTIVE: To quantify the intracycle variation in markers of ovarian reserve measured by antral follicle counts stratified by size using three-dimensional (3D) ultrasound and anti-Müllerian hormone (AMH) in women with normal menstrual cycles. METHODS: Healthy volunteers with normal menstrual cycles were prospectively recruited. Three-dimensional (3D) ultrasound examination and blood test were performed in early (F1) and mid-follicular (F2) phases and in periovulatory (PO) and luteal (LU) phases of one menstrual cycle. Antral follicles were measured using 'sonography-based automated volume calculation' with post processing (SonoAVC) and ovarian volume was measured using Virtual Organ Computer-aided AnaLysis (VOCAL). Blood serum was processed for hormonal assays including AMH, follicle stimulating hormone (FSH), luteinizing hormone (LH) and estradiol. Repeated-measures analysis was used to examine the variance in markers of ovarian reserve in different phases of one menstrual cycle. RESULTS: A total of 36 volunteers were included in the final analysis, of whom 34 attended all four visits. Repeated-measures analysis showed a significant variation in total antral follicle count (AFC) (P < 0.001). However, on stratifying the antral follicles according to size using SonoAVC, a non-significant variation (P = 0.382) was seen in small AFC (≤ 6.0 mm) and a significant variation (P < 0.001) was seen in large AFC (> 6.0 mm). The ovarian volume showed a significant intracycle variation (P < 0.001). A small but significant intracycle variation was noted in AMH (P = 0.041) and a significant variation was seen in levels of serum FSH, LH and estradiol (P < 0.05). CONCLUSION: Small antral follicles (≤ 6.0 mm) measured using 3D ultrasound and AMH show little intracycle variation and perhaps should be evaluated when predicting ovarian reserve independent of menstrual cycle.

Quantifying effect of combined oral contraceptive pill on functional ovarian reserve as measured by serum anti-Müllerian hormone and small antral follicle count using three-dimensional ultrasound.

OBJECTIVES: Oral contraceptive pills suppress the hypothalomo-pituitary axis, which can affect the ultrasound and endocrine markers used to examine ovarian reserve. The objective of this study was to quantify the ultrasound and endocrine markers of functional ovarian reserve in women using a combined oral contraceptive pill (COCP) for more than a year. METHODS: This was a prospective case-control study involving healthy volunteers: 34 women using for more than a year a COCP with hormone-free interval (HFI) were compared to 36 normo-ovulatory, age-matched controls who had not used hormonal contraception within the last year. Volunteers using a COCP underwent a 3D ultrasound examination and had a blood sample taken within the first 4 days of active pill ingestion and those in the control group had the scan and blood test in the early follicular phase (days 2-5) of menstrual cycle. The main outcome measure was the difference in antral follicle counts stratified according to size and anti-Müllerian hormone (AMH), follicle-stimulating hormone (FSH), luteinizing hormone (LH) and estradiol (E2) levels. RESULTS: There were no significant differences in the number of small antral follicles measuring 2-6 mm. The COCP group had significantly fewer antral follicles measuring ≥ 6 mm (P < 0.001) and had significantly smaller ovaries (P < 0.001), which also had lower vascular indices than the control group (P < 0.05). While serum FSH, LH and E2 levels were significantly lower in the COCP group (P < 0.05), there was no significant difference in serum AMH levels between the two groups. CONCLUSIONS: Prolonged use of COCP suppressed pituitary gonadotropins and antral follicle development beyond 6 mm, but had no effect on levels of serum AMH and number of small antral follicles.

Three-dimensional Doppler evaluation of single spherical samples from the placenta: intra- and interobserver reliability.

OBJECTIVE: To evaluate the intra- and interobserver reliability of assessment of three-dimensional power Doppler (3D-PD) indices from single spherical samples of the placenta. METHODS: Women with singleton pregnancies at 24-40 weeks' gestation were included. Three scans were independently performed by two observers; Observer 1 performed the first and third scan, intercalated by the scan of Observer 2. The observers independently analyzed the 3D-PD datasets that they had previously acquired using four different methods, each using a spherical sample: random sample extending from basal to chorionic plate; random sample with 2 cm(3) of volume; directed sample to the region subjectively determined as containing more color Doppler signals extending from basal to chorionic plate; or directed sample with 2 cm(3) of volume. The vascularization index (VI), flow index (FI) and vascularization flow index (VFI) were evaluated in each case. The observers were blinded to their own and each other's results. Additional evaluation was performed according to placental location: anterior, posterior and fundal or lateral. Intra- and interobserver reliability was assessed by intraclass correlation coefficients (ICC). RESULTS: Ninety-five pregnancies were included in the analysis. All three placental 3D-PD indices showed only weak to moderate reliability (ICC < 0.66 and ICC < 0.48, intra- and interobserver, respectively). The highest values of ICC were observed when using directed spherical samples from basal to chorionic plate. When analyzed by placental location, we found lower ICCs for lateral and fundal placentae compared to anterior and posterior ones. CONCLUSION: Intra- and interobserver reliability of assessment of placental 3D-PD indices from single spherical samples in pregnant women greater than 24 weeks' gestation is poor to moderate, and clinical usefulness of these indices is likely to be limited.

Reproductive outcomes in women with congenital uterine anomalies: a systematic review.

OBJECTIVE: Congenital uterine anomalies are common but their effect on reproductive outcome is unclear. We conducted a systematic review to evaluate the association between different types of congenital uterine anomaly and various reproductive outcomes. METHODS: Searches were performed using MEDLINE, EMBASE, the Cochrane Library and Web of Science. The Newcastle-Ottawa Quality Assessment Scale was used for quality assessment. Uterine defects were grouped into arcuate uteri, canalization defects (septate and subseptate uteri) and unification defects (unicornuate, bicornuate and didelphys uteri). Pooled risk ratios (RR) with 95% confidence intervals (CI) were computed using random effects models. RESULTS: We identified nine studies comprising 3805 women. Meta-analysis showed that arcuate uteri were associated with increased rates of second-trimester miscarriage (RR, 2.39; 95% CI, 1.33-4.27, P = 0.003) and fetal malpresentation at delivery (RR, 2.53; 95% CI, 1.54-4.18; P < 0.001). Canalization defects were associated with reduced clinical pregnancy rates (RR, 0.86; 95% CI, 0.77-0.96; P = 0.009) and increased rates of first-trimester miscarriage (RR, 2.89; 95% CI; 2.02-4.14; P < 0.001), preterm birth (RR, 2.14; 95% CI, 1.48-3.11; P < 0.001) and fetal malpresentation (RR, 6.24; 95% CI, 4.05-9.62; P < 0.001). Unification defects were associated with increased rates of preterm birth (RR, 2.97; 95% CI, 2.08-4.23; P < 0.001) and fetal malpresentation (RR, 3.87; 95% CI, 2.42-6.18; P < 0.001). CONCLUSIONS: Canalization defects reduce fertility and increase rates of miscarriage and preterm delivery. None of the unification defects reduces fertility but some are associated with miscarriage and preterm delivery. Arcuate uteri are specifically associated with second-trimester miscarriage. All uterine anomalies increase the chance of fetal malpresentation at delivery.

Embryo volume measurement: an intraobserver, intermethod comparative study of semiautomated and manual three-dimensional ultrasound techniques.

OBJECTIVE: To compare the reliability of our recently introduced technique for first-trimester embryo volume measurement, the 'semiautomated technique' using both Virtual Organ Computer-aided AnaLysis (VOCAL(™) ) and Sonography-based Automated Volume Count (SonoAVC) with a manual technique using VOCAL alone. METHODS: Seventy-four subjects with viable, singleton first-trimester in-vitro fertilization (IVF) pregnancies were recruited. Each subject underwent transvaginal sonography, at which a three-dimensional ultrasound dataset of the entire gestational sac was acquired. Each embryo volume was measured by two techniques, each performed twice. In the semiautomated technique VOCAL was used to calculate the volume of the gestational and yolk sacs, and SonoAVC was used to quantify the fluid volume within the amniotic and extra-amniotic cavities. Embryo volume was calculated by subtracting the sum of yolk sac, amniotic and extra-amniotic fluid volumes from gestational sac volume. In the manual technique, VOCAL was used to delineate the entire embryo using 9° rotations. Reliability was assessed using limits of agreement and intraclass correlation coefficient. RESULTS: Datasets were included from 52 eligible subjects. Median gestational age was 7 + 4 weeks; median crown-rump length (CRL) was 13 (range, 2-29) mm; and median embryo volume was 1.8 (range, 0.03-8.1) cm(3) using the semiautomated technique and 0.7 (range 0.009-3.6) cm(3) using the manual technique. There was a significant discrepancy in the volumes measured by the two different techniques. Assessment of the limits of agreement suggested that the semiautomated technique (-15.8% to 14.0% of the mean embryo volume) was more reliable than was the manual technique (-22.4% to 26.6%). CONCLUSION: The semiautomated technique is more reliable than is the manual technique for embryo volume measurement. However, the discrepancy in measurements between the two methods raises concerns over the validity of the semiautomated technique that require further investigation.

A standardized measurement technique may improve the reliability of measurements of endometrial thickness and volume.

OBJECTIVE: To investigate whether standardization of the multiplanar view (SMV) when evaluating the uterus using three-dimensional ultrasonography (3D-US) improves intra- and interobserver reliability and agreement with regard to endometrial measurement. METHODS: Two-dimensional (2D) and 3D-US was used to measure endometrial thickness by two observers in 30 women undergoing assisted reproduction treatment. Endometrial volume was measured with Virtual Organ Computer-aided AnaLysis (VOCAL(™)) in the longitudinal (A) and coronal (C) planes using an unmodified multiplanar view (UMV) and a standardized multiplanar view (SMV). Measurement reliability was evaluated by intraclass correlation coefficient (ICC) and agreement was examined using Bland-Altman plots with limits of agreement (LoA). The ease of outlining the endometrial-myometrial interface was compared between the A- and C-planes using subjective assessment. RESULTS: Endometrial volume measurements using the SMV and A-plane were more reliable (intra- and interobserver ICCs, 0.979 and 0.975, respectively) than were measurements of endometrial thickness using 2D-US (intra- and interobserver ICCs, 0.742 and 0.702, respectively) or 3D-US (intra- and interobserver ICCs, 0.890 and 0.784, respectively). The LoAs were narrower for SMV than for UMV. Reliability and agreement were not much different between the A- and C-planes. However the observers agreed that delineating the endometrial-myometrial interface using the A-plane was easier (first and second observer, 50.0 and 46.7%, respectively) or 'comparable' (50 and 53.3%, respectively), but never more difficult than using the C-plane. CONCLUSIONS: Endometrial volume measurements are more reliable than endometrial thickness measurements and are best performed using SMV and the A-plane.

Prevalence of uterine anomalies and their impact on early pregnancy in women conceiving after assisted reproduction treatment.

OBJECTIVE: To estimate the prevalence of congenital uterine anomalies in subfertile women and to evaluate their influence on early pregnancy following assisted reproduction treatment (ART). METHODS: We prospectively recruited 1402 subjects undergoing ART over a period of 5 years from 2005 to 2009. Three-dimensional transvaginal sonography was performed in the early follicular phase of the menstrual cycle (days 2-5) and repeated in the late follicular phase (days 10-14) if the shape of the uterine cavity could not be assessed at the first scan. All subjects who conceived following ART were followed up to 12 weeks' gestation. Chi-square test was used to compare the pregnancy rates and miscarriage rates between women shown to have uterine anomalies and those with a normal uterus. RESULTS: One thousand three hundred and eighty-five subjects were included for final analysis after excluding 17 subjects in whom a definitive diagnosis could not be made. While 1201 (86.7%) subjects had a normal uterine cavity, uterine anomalies were demonstrated in 184 (13.3%) subjects. Arcuate uteri represented the commonest anomaly (n = 164 (11.8%)) followed by septate (n = 7 (0.5%)), unicornuate (n = 6 (0.4%)), subseptate (n = 5 (0.4%)), bicornuate (n = 1 (0.1%)) and T-shaped uteri (n = 1 (0.1%)). A total of 440 subjects who underwent ART were followed up. The pregnancy rates in women with arcuate uteri (36/66 (54.5%)) and major uterine anomalies (7/10 (70.0%)) were statistically similar (P = 0.09 and P = 0.11, respectively) to that of the matched controls with normal uteri (158/364 (43.4%)). While first-trimester miscarriage rates were similar (P = 0.81) between the control group (20/158 (12.7%)) and women with arcuate uteri (5/36 (13.9%)), women with major uterine anomalies experienced a higher miscarriage rate (3/7 (42.9%); P = 0.05). CONCLUSIONS: Women who are referred for ART have a high prevalence of congenital uterine anomalies, the most common anomaly being an arcuate uterus. These anomalies are not associated with a reduction in pregnancy rates following ART. However, while the arcuate uterus was not associated with an increase in first-trimester miscarriage, major uterine anomalies seemed to increase the risk of first-trimester miscarriage.

Quantitative analysis of antral follicle number and size: a comparison of two-dimensional and automated three-dimensional ultrasound techniques.

OBJECTIVES: To compare two-dimensional (2D) ultrasound imaging with automated three-dimensional (3D) ultrasound imaging for the measurement of antral follicle number and size. METHODS: Twenty-four subjects aged < 40 years underwent transvaginal ultrasound examination (Voluson E8) in the early follicular phase of the menstrual cycle. A 2D ultrasound scan of both ovaries was performed; each antral follicle was identified and then measured by taking the mean of two diameters. A 3D ultrasound dataset of both ovaries was then acquired and analyzed using Sonography-based Automated Volume Count (SonoAVC). The time taken to measure the size of all antral follicles in both ovaries was recorded in seconds for each technique. Antral follicle size was recorded to the nearest millimeter and counts for each 1-mm group were obtained. Antral follicle counts were also grouped according to five predefined size categories: 2.0-5.0 mm, 2.0-6.0 mm, 2.0-8.0 mm, 2.0-9.0 mm and 2.0-10.0 mm. Limits of agreement (LOA) and a paired t-test or Wilcoxon signed ranks test were used to analyze the data depending on their distribution. RESULTS: When antral follicle numbers were compared for each 1-mm follicle size group, 2D ultrasound imaging recorded more follicles measuring 3.0-3.99 mm (mean +/- SD, 4.11 +/- 3.70 vs. 2.63 +/- 2.31; P = 0.019) and 4.0-4.99 mm (mean +/- SD, 4.63 +/- 4.86 vs. 2.68 +/- 2.89; P = 0.013) than did SonoAVC. LOA were widest with follicles measuring 3.0-3.99 mm (LOA, 6.38 and -3.43) and 4.0-4.99 mm (LOA, 7.99 and -4.09). The antral follicle count in each of the five predefined size categories was significantly lower with SonoAVC than with 2D ultrasound imaging (P < 0.05). SonoAVC took significantly less time to measure the size and record the number of antral follicles than did 2D ultrasound imaging (mean +/- SD, 132.05 +/- 56.23 s vs. 324.47 +/- 162.22 s; P < 0.001). CONCLUSIONS: Fewer antral follicles are evident overall when SonoAVC is used to analyze 3D ultrasound data. The clinical significance of this remains to be determined but the automated technique is significantly quicker than is making measurements using 2D ultrasound imaging.

Quantitative three-dimensional power Doppler angiography: a flow-free phantom experiment to evaluate the relationship between color gain, depth and signal artifact.

OBJECTIVES: To evaluate the presence of false flow three-dimensional (3D) power Doppler signals in 'flow-free' models. METHODS: 3D power Doppler datasets were acquired from three different flow-free phantoms (muscle, air and water) with two different transducers and Virtual Organ Computer-aided AnaLysis was used to generate a sphere that was serially applied through the 3D dataset. The vascularization flow index was used to compare artifactual signals at different depths (from 0 to 6 cm) within the different phantoms and at different gain and pulse repetition frequency (PRF) settings. RESULTS: Artifactual Doppler signals were seen in all phantoms despite these being flow-free. The pattern was very similar and the degree of artifact appeared to be dependent on the gain and distance from the transducer. False signals were more evident in the far field and increased as the gain was increased, with false signals first appearing with a gain of 1 dB in the air and muscle phantoms. False signals were seen at a lower gain with the water phantom (-15 dB) and these were associated with vertical lines of Doppler artifact that were related to PRF, and disappeared when reflections were attenuated. CONCLUSIONS: Artifactual Doppler signals are seen in flow-free phantoms and are related to the gain settings and the distance from the transducer. In the in-vivo situation, the lowest gain settings that allow the detection of blood flow and adequate definition of vessel architecture should be used, which invariably means using a setting near or below the middle of the range available. Additionally, observers should be aware of vertical lines when evaluating cystic or liquid-containing structures.

The predictive value of the automated quantification of the number and size of small antral follicles in women undergoing ART.

BACKGROUND: Sono-automatic volume calculation (SonoAVC) automatically identifies and measures the dimensions of hypoechogenic areas within datasets acquired using three-dimensional ultrasound. The objective of this study was to evaluate the predictive value of automated antral follicle counts according to their relative sizes in women undergoing assisted reproduction treatment (ART). METHODS: A total of 156 subjects aged < or =40 years with a baseline FSH < or =15 IU that undergo their first cycle of ART were prospectively recruited. SonoAVC was used to measure the datasets and record the number of antral follicles measuring < or =9 mm in diameter. These follicles were then grouped into subsets according to their relative sizes: < or =2.0, 2.1-4.0, 4.1-6.0, 6.1-8.0 and 8.1-9.0 mm. The primary outcome was viable pregnancy confirmed on ultrasound 5 weeks following embryo transfer. RESULTS: A total of 142 subjects were included for analysis of primary end-point. Those subjects who conceived had significantly more antral follicles measuring < or =2 (P = 0.041) and 2.1-4.0 mm (P < 0.001) than those who had unsuccessful treatment. There were no significant differences between the groups in the number of antral follicles measuring 4.1-6.0 (P = 0.191), 6.1-8.0 (P = 0.203) and 8.1-9.0 mm (P = 0.601). Multiple logistic regression showed that antral follicles measuring 2.1-4.0 mm were an independent predictor of pregnancy [Exp(B) = 1.234, 95% CI = 1.092-1.491; P = 0.004; AUC = 0.693]. CONCLUSION: SonoAVC provides automated measures of antral follicle number and size. Using this technique, the number of antral follicles measuring 2.1-4.0 mm in diameter is an independent, significant predictor of pregnancy following in vitro fertilization treatment.

Can quantitative three-dimensional power Doppler angiography be used to predict ovarian hyperstimulation syndrome?

OBJECTIVE: To test the hypothesis that ovarian vascularity is increased in women developing ovarian hyperstimulation syndrome (OHSS) and to assess its value as a predictor of OHSS during in-vitro fertilization (IVF). METHODS: 118 subjects undergoing their first cycle of IVF had a three-dimensional (3D) transvaginal ultrasound scan in the early follicular phase of the menstrual cycle preceding IVF treatment. 18 of them developed moderate or severe OHSS and 100 subjects had normal ovarian response. Antral follicle count, ovarian volume, and ovarian vascularity (vascularization index (VI), flow index (FI) and vascularization flow index (VFI)) were compared between OHSS and control groups. Multiple regression analysis was used to assess the predictive value of these variables against age, body mass index and basal follicle-stimulating hormone level for the development of OHSS. RESULTS: The ovarian blood flow indices VI (11.1 +/- 11.6 vs. 8.6 +/- 7.3%; P = 0.23), FI (38.0 +/- 4.8 vs. 38.0 +/- 5.5; P = 0.95) and VFI (4.2 +/- 3.3 vs. 3.5 +/- 3.1; P = 0.40) were similar in the OHSS group and the normal responders. While antral follicle count was significantly higher in women developing OHSS (33.0 +/- 15.1) than in the control group (19.2 +/- 9.9, P < 0.001), ovarian volume did not differ between the two groups (10.6 +/- 3.8 vs. 8.9 +/- 4.8 cm(3), respectively, P = 0.11). On multiple regression analysis, antral follicle count was the only significant predictor of OHSS (P < 0.01). CONCLUSIONS: Women developing OHSS during IVF do not demonstrate an increased ovarian blood flow as measured by 3D ultrasound but do have a significantly higher antral follicle count, which is the only significant predictor of OHSS.

Intraobserver and interobserver reliability of automated antral follicle counts made using three-dimensional ultrasound and SonoAVC.

OBJECTIVE: To assess the reliability of automated measurements of the total antral follicle count (AFC) made using Sono-Automatic Volume Count (SonoAVC), and to compare these to two-dimensional (2D) and manual three-dimensional (3D) techniques. METHODS: Fifty-five subjects aged under 40 years who had 3D transvaginal ultrasound examination in the early follicular phase of their menstrual cycle were prospectively recruited. 3D datasets were acquired and subsequently analyzed. The total AFC (2-10 mm antral follicles) was calculated by two observers using three independent methods: 2D real-time equivalent (2D-RTE), 3D manual multiplanar view (3D-MPV), and SonoAVC. For measurements made using SonoAVC, the initial automated count (sAVC-AA) was recorded and postprocessing (sAVC-PP) then applied to identify follicles that had been missed or incorrectly included. Intraclass correlation and limits of agreement were used to evaluate the methods. RESULTS: The intra- and interobserver reliability of measurements of total AFC was best with SonoAVC with postprocessing followed by 3D-MPV and 2D-RTE. The initial count calculated by sAVC-AA missed follicles and this was reflected in the significantly lower mean total AFC (6.51 +/- 4.79) than that made after postprocessing techniques (sAVC-PP, 18.42 +/- 10.53, P < 0.001; 3D-MPV, 19.38 +/- 10.85, P < 0.001; and 2D-RTE, 19.26 +/- 10.55, P < 0.001). The mean total AFC became more comparable with postprocessing (sAVC-PP) but still remained significantly lower than counts made with 2D-RTE and 3D-MPV (P < 0.05). CONCLUSION: SonoAVC with postprocessing is a reliable method for measuring total AFC. It takes longer to perform, because of the need for postprocessing, and obtains values that are lower than those obtained by the 2D and 3D-MPV techniques. However, the AFC obtained by sAVC-PP is likely to be lower because this method measures and color codes each follicle preventing recounting.

Changes in myometrial 'perfusion' during normal labor as visualized by three-dimensional power Doppler angiography.

OBJECTIVES: Myometrial contractions are one of the most important aspects of effective labor. For cells within the myometrium to work efficiently they need to be well oxygenated and this requires an adequate blood supply. This study used quantitative three-dimensional (3D) power Doppler angiography to calculate the percentage change in myometrial blood flow during a relaxation-contraction-relaxation cycle of active labor. METHODS: Transabdominal 3D power Doppler ultrasound imaging was used to acquire volumetric data during the first stage of spontaneous labor in 20 term, nulliparous women. 3D datasets were acquired during a single cycle of uterine relaxation, contraction and subsequent relaxation for each subject. The resultant datasets were analyzed independently by two investigators on two occasions using Virtual Organ Computer-aided AnaLysis to define a volume of interest within the myometrium; the power Doppler signal within this volume was quantified to provide 3D indices of vascularity: vascularization index (VI), flow index (FI) and vascularization flow index (VFI). The percentage change in these indices, during a uterine contraction, was calculated from the baseline value during the initial uterine relaxation phase (taken as a maximum of 100%). RESULTS: Myometrial blood flow fell significantly during the uterine contraction and returned during the subsequent relaxation phase of the cycle (P < 0.001 for VI and VFI, P = 0.002 for FI). From the initial baseline relaxation value, VI dropped to 43.9%, FI to 85.5% and VFI to 40.8% during uterine contraction, and returned to 86.7%, 98.1% and 89.1%, respectively, during the subsequent relaxation. The intraclass correlation coefficients in blood flow measurements of 0.982-0.999 between the two investigators were indicative of good interobserver reliability. CONCLUSIONS: This study confirms that myometrial perfusion, as measured by quantitative 3D power Doppler angiography, significantly falls during uterine contractions, returns during the subsequent relaxation phase, and can be quantified reliably from stored datasets. Further work is now required to establish clinical applicability for this non-invasive investigation.

In vitro dual perfusion of human placental lobules as a flow phantom to investigate the relationship between fetoplacental flow and quantitative 3D power doppler angiography.

Flow phantoms have been used to investigate and quantify three-dimensional power Doppler data but this is the first study to use the in vitro, dual perfused, placental perfusion model. We used this model to investigate and quantify the effect of variation in fetal-side flow rates and attenuation on 3D power Doppler angiography. Perfusion of a placental lobule was commenced within 30 min of delivery and experimentation was successful in 8 of the 18 placenta obtained. Fetal and maternal perfusate was modified Earle's bicarbonate buffer which, following equilibration, was supplemented on the fetal side with whole heparinised cord blood. Imaging was performed with a Voluson-i ultrasound machine. A 'vascular biopsy' the thickness of the placental lobule was defined and signal quantified within using VOCAL (GE Medical Systems, Zipf, Austria). Three vascular indices are generated: vascularisation index (VI) defined as the percentage of power Doppler data within a volume of interest; flow index (FI), the mean signal intensity of the power Doppler information; and vascularisation flow index (VFI), a combination of both factors derived through their multiplication. Attenuation was investigated in this model with the addition of tissue mimic blocks. Our results showed a predictable relationship between flow rates and the vascular indices VI and VFI. However the FI was a less reliable predictor of flow; thus it should be interpreted with caution. The power Doppler signal was markedly affected by attenuation leading to a complete loss of information at a depth of 6 cm in the model used. In conclusion this model can be adapted to provide a phantom to analyse and quantify 3D power Doppler signals and demonstrates that vascular indices within a tissue remain related to volume flow. This model provides further evidence that depth dependent attenuation of signal needs to be accounted for in any in vivo work where the probe is not in direct contact with the tissue of interest.

Evaluation of the effect of machine settings on quantitative three-dimensional power Doppler angiography: an in-vitro flow phantom experiment.

OBJECTIVES: Three-dimensional (3D) ultrasound is being used increasingly to acquire and subsequently quantify power Doppler data within the clinical setting. One proprietary software package calculates three 3D vascular indices: the vascularization index (VI), the flow index (FI), and the vascularization flow index (VFI). Our aim was to evaluate how different settings affect the Doppler signal in terms of its quantification by these three indices within a 3D dataset. METHODS: A computer-driven 'flow phantom' was used to continuously pump a nylon particle-based blood mimic (Orgasol(trade mark)) around a closed system through a C-flex(trade mark) tube embedded in an agar-based tissue mimic. The test tanks were insonated with a modified 3D transvaginal 4-8-MHz ultrasound transducer (V530D) and power Doppler data were acquired over a series of different settings. Each experiment involved the manipulation of just one Doppler setting in order to study it in isolation. RESULTS: As expected, all of the power Doppler settings, when altered, were found to effect significant changes (P < 0.05) in the VI, FI and VFI. The gain and signal power had the greatest effect, producing no Doppler signals at the lowest settings and the highest recordable indices at the maximum settings. The pulse repetition frequency (PRF) was the next most influential setting but a Doppler signal was seen and measurable at all of the different settings. The other Doppler settings had a much less profound effect on the vascular indices, with subtle but significantly different measures across the full range of settings. The speed of data acquisition was also found to affect the vascular indices, all of which were reduced when the fast mode was used although the only significant effect was on the VFI. CONCLUSIONS: The VI, FI and VFI are all affected significantly by variations in power Doppler settings and by the speed of acquisition. The gain and signal power have the greatest effect on the power Doppler signal, followed closely by the PRF. The other settings and speed of acquisition also influence the signal, but to a much lesser degree. It is essential to maintain Doppler settings if any meaningful comparisons are to be made within and between subjects.