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Pulsed Doppler and Color Duplex Sonography in the Assessment of Tubal Patency
Fig. 7.5 Original image showing contrast medium in the uterine fun-
dus. The sample volume is positioned over the intramural segment of the left tube. The lower part of the image shows the Doppler shift as­sociated with contrast arrival and decreased outflow due to partial tubal obstruction.
7
Fig. 7.7 Original images.
a Color Doppler appearance of contrast medium entering the proxi-
mal and middle thirds of the left tube.
Fig. 7.6 Original image showing contrast medium in the intramural part of the left tube. The proximal and middle thirds of the tube can be identified in the high-resolution B-mode image.
b Outflow of contrast medium from the fimbriated end of the left tube (turbulent zones are encoded in blue and red next to a polyfollic­ular left ovary).
72
Fig. 7.8 Original images.
a The sample volume is positioned over the proximal third of the right
tube.
b Initial inflow of thecontrast medium is marked by strong turbulence in the tubal lumen.
Fig. 7.8c Maximum turbulent flow in the tubal lumen (red = flow toward the transducer).
Assessment of Tubal Patency
Table 7.2 Results of Doppler ultrasound hysterosalpingography of
404 tubes in 210 patients
a Proximal segments
Diagnostic method Patent Obstruction
Partial Complete
B-mode 373 (92%) 31 (8 %) B-mode and pulsed
Doppler
b Distal segments
Diagnostic method Patent Obstruction
B-mode 292 (72 %) 112 (28 %) B-mode and pulsed
Doppler
72% (292 tubes) as patent and 28 % (112 tubes) as obstructed. Subsequent Doppler analysis indicated patency in 73% (297 tubes), partial obstruction in 7 % (30 tubes), and complete ob­struction in 20 % (77 tubes).
350 (87%) 24 (6%) 30 (7%)
Partial Complete
297 (73%) 30 (7%) 77 (20%)
Infertility Evaluation and Assisted Reproduction
Fig. 7.8d Outflow of contrast medium from the proximal into the distal segment.
Fig. 7.8e Contrast spillage from the fimbriated end of the right tube.
Results
Comparison of B-mode and Doppler. A total of 404 fallopian
tubes in 210 patients were sonographically assessed for patency using B-mode and Doppler flow analysis. The results
were determined separately for the proximal and distal tubal segments (Table 7. ment, patency was diagnosed in 92% (373 tubes) and partial or complete obstruction in 8 % (31 tubes). When Doppler analysis
was added, the results were more differentiated: 87% of the tubes (350) were patent, 6 % (24 tubes) were partially ob­structed, and 7% (30 tubes) were completely obstructed. Using B-mode in the distal tubal segment (Table 7.
2). Using B-mode in the proximal tubal seg-
2b), we classified
Comparison with laparoscopy. These sonographic findings in
404 tubes werechecked by laparoscopyin 62% of all cases (252
tubes) (Table 7.
3). To determine the rate of agreement between
the two methods, we subdivided the whole population (404 tubes) into three sonographic groups:
Patent (297 tubes)
Partial obstruction (35 tubes)
Complete obstruction (72 tubes)
The rates of agreement between sonographic and laparoscopic findings for the three groups were 95%, 95 %, and 84%. Between 60% and 65% of the tubes had been examined endoscopically in all three of the groups.
The overall rate of agreement for the three subgroups was
92% (233 of 252 tubes examined).
Treatment. The patients diagnosed by ultrasound tubal imag­ing were either referred for ovarian stimulation therapy, fol­lowed in some cases by intratubal gamete transfer,or admitted to the IVF program.
Table 7.3 Results of laparoscopic control examinations compared
with Doppler ultrasound hysterosalpingography of 404 tubes in 210 patients, rates of agreement between ultrasound and endoscopy
Result Total Checked by
laparoscopy
Patent (297) 184 (62 %) 174 (95 %) Partially
obstructed Completely
obstructed All categories (404) 252 (62%) 233 (92%)
(35) 21 (60 %) 20 (95 %)
(72) 47 (65 %) 39 (84 %)
Agreement between ultrasound and laparoscopy
73
Pulsed Doppler and Color Duplex Sonography in the Assessment of Tubal Patency
74
Table 7.4 Side-effects associated with ultrasound contrast imaging of the fallopian tubes
Side-effects Patients (210 total)
Vasovagal reaction, pain 14 (7%) Adnexitis 4 (2 %) Allergic reactions 0
Side-effects. Table 7.4 lists the side-effects that were registered in the study. Fourteen of the 210 patients (7%) experienced a vasovagal reaction after several milliliters of the absorbable contrast medium had been injected, necessitating discon­tinuation of the examination. Four patients (2 %) developed mild pelvic inflammatory disease, which resolved quickly in response to antibiotic treatment. Other complications, es­pecially allergic reactions, were not observed.
Discussion of the Value of the Test Procedures
B-mode imaging. Advances in ultrasound technology, includ-
7
ing the development of high-resolution endovaginal trans­ducers, have enabled anatomically precise visualization of the female internal genital organs gans of interest, these transducers can be operated at higher frequencies to provide better resolution of very small details. As a result, we can evaluate the uterus and ovaries and appre­ciate their cycle-dependent changes on plain ultrasound im-
1, 8
ages
. B-mode is a sensitive, noninvasive, ambulatory method for the detection of internal genital malformations in infertile patients and in women with palpable stage of the infertility workup, ultrasound imaging and es­pecially transvaginal sonography can already provide an effec­tive, noninvasive screening method (sensitivity 43%, speci­ficity 98%)
12
. Menstrual function studies such as folliculometry and endometrial testing have also become established ultra­sound procedures in infertility patients
Radiographic hysterosalpingography and laparoscopic chro­mopertubation. Tubal infertility has a prevalence of 20–30%,
which is currently increasing owing to the rising incidence of salpingitis, ectopic pregnancy, and their conservative manage-
5
ment
. In the past, the only reliable and established methods available to the fertility specialist in the pretherapeutic inves­tigation of an unexplained tubal factor were radiographic hys­terosalpingography (HSG) and laparoscopic
11, 15 , 17
tion
Both of these methods have drawbacks such as radiation exposure, possible contrast allergies, pain, invasive­ness, and risks relating to general anesthesia, bleeding, and in­fection. There has been a pressing need for a noninvasive, risk­free screening method for tubal patency that can be performed in an outpatient setting
Abdominal sonography after uterine fluid instillation. Plain ul­trasound examination of the lesser pelvis has been unable to provide information on the dynamics of tubal transit. Only ab­normal fluid collections (hydrosalpinx, sactosalpinx) could be visualized. Richman et al. (1984) and Randolph et al. (1986) re-
2
. With their proximity to the or-
abnormalities
1, 8
.
12
chromopertuba-
. At this
ported on the assessment of tubal patency with abdominal ul­trasound following intrauterine fluid instillation
13, 14
. This method was basically indirect, however, relying on the detec­tion of free fluid in the cul-de-sac to confirm tubal patency. It also required the use of large amounts of saline solution (up to 200 ml), leading to an increased risk of infection. The examina­tion was performed under general anesthesia before a planned laparoscopy or HSG.
Transvaginal hysterocontrast sonography. Deichert et al. (1988, 1989) introduced hysterocontrast sonography as a new diagnostic procedure for the differentiation of intrauterine and myometrial findings
4
. They used saline solution for examina­tion of the uterine cavity and a galactose microparticle suspen­sion for contrast imaging of the fallopian tubes. These studies, unlike our own, were performed under general anesthesia. Turbulent flow within the tube, contrast spillage from the fimbriated end, and increased fluid in the cul-de-sac were the criteria used for the B-mode assessment of tubal patency
5
. The authors reported 65% agreement with laparoscopy and radio­graphic HSG, with a 33% rate of partial agreement.
Doppler sonography. After Fitzgerald and Drumm introduced the Doppler principle into obstetric medicine in 1977
6
, Taylor et al. (1985) were the first to perform blood flow measure­ments of the ovarian and uterine arteries during the menstrual cycle, thus applying the principle to the field of reproductive medicine
16
. They used abdominal PW sector transducers as well as endovaginal CW transducers under general anesthesia before a scheduled laparotomy. Since the early 1990s, authors have reported on a varietyof results using transvaginalB-mode and Doppler imaging techniques in almost all areas of repro­ductive and prenatal medicine
9c
. With the high-resolution en­dovaginal transducers currently available, flow signals can be selectively recorded from the lesser pelvis using spectral and color Doppler techniques down to penetration depths of 10–20cm. But because some internal genital structures (fal­lopian tubes) generally cannot be evaluated in their anatomy or function by plain imaging, it is necessary to incorporate con­trast administration into the ultrasound protocol
3a
.
Doppler sonography with contrast medium. After our group had established B-mode contrast visualization of the fallopian tubes in good agreement with classic methods
10
, we attempted to achieve an even more accurate, dynamic, noninvasive assessment of tubal patency by combining this technique with the pulsed Doppler
principle
9a
. The examination can be done on an ambulatory basis, but for safety reasons we combined it with one night of hospital observation in the early phase of the study.The method is wellaccepted by patients as it does not in­volve premedication, general anesthesia, or pain. If the cervical canal is stenotic or poorly estrogen-primed, or if the patient has a uterine position anomaly, the cervix may be grasped with a tenaculum to straighten the uterus, or a special intrauterine catheter may be used that has greater stability and a smaller lumen (Zinnanti Instruments, Chatsworth, CA, USA).
Contrast medium. The absorbable contrast medium incited a vasovagal response in a few patients (7 %), requiring the exami­nation to be terminated. Four patients developed mild pelvic

Summary

inflammatory disease, which responded well to antibiotics. We observed no serious allergic reactions like those associated
with iodinated contrast media.
Standard protocol. The examination should follow a standard protocol using designated planes of section. Following contrast instillation, the uterine cavity is first surveyed in longitudinal and transverse sections. Gross uterine malformations and ab­normalities (bicornuate uterus, subseptate uterus, leiomy­omas, etc.) should be excluded. This is followed by a dynamic examination of both fallopian tubes, starting with a B-mode analysis and proceeding to Doppler spectral analysis. The tubes cannot be fully visualized with B-mode because of their tor­tuosity, but portions of the tubes can be identified, even pe­ripherally, and the flow pattern of the contrast medium can be analyzed by proper placement of the sample volume. This can significantly increase the quality and information content of the B-mode
examination
9b
(Table 7.2).
Color Doppler analysis. Color Doppler analysis can further add to the diagnostic capabilities of the ultrasound examination
3, 7
Multiple color boxes are placed all over the B-mode image, al­lowing flow to be measured from one pixel to the next over the entire sectional image. Flow toward the transducer is encoded in red, flow away from the transducer in blue. Turbulent flow produces a mixed pattern of green and yellow pixels. In con­trast to unidimensional analysis with spectral Doppler, flow phenomena can be imaged simultaneously over the entire scanned region. This requires a considerably more time-con­suming adjustment of the color sensitivity of the ultrasound system, and the increase in examination time can be justified only in cases where the spectral Doppler findings are equivo-
cal. Moreover, the contrast visualization of flow in the tubal segments with a 9 MHz transducer can show details in the B­mode image that are comparable to those revealed by color Doppler. Generally, however, only color-flow imaging can demonstrate fluid spillage from the fimbriated end of the tube.
With the Doppler technique described above, it is not possible to evaluate the mobility of the infundibulum or peri­tubal region, even when ultrasound confirms tubal patency.
This explains why postsonographic laparoscopic findings were used in admitting patients to the IVF program. The 92% con­cordance between the studies (233 of 252 tubes examined in 210 patients) (Table 7. ported in a review by Campbell et al.
3) agrees with the rates of 80–91% re-
3a
and identifies the method as a serious alternative to the invasive procedures of radiographic HSG and diagnostic laparoscopy.
Comments. The assessment of tubal patency by transvaginal pulsed Doppler scanning and in selected cases by color duplex sonography provides an easy-to-use, noninvasive screening method for excluding tubal obstruction. It is superior to radio-
graphic HSG in the concomitant evaluation of uterine malfor-
.
mations and anatomical variants. It shows good agreement
with the classic diagnostic tests and is potentially safer owing to high patient tolerance of the contrast medium. Done as an outpatient procedure with no premedication or general an­esthesia, it is less costly and more comfortable for the patients.
If the findings are equivocal, the classic procedures of radio-
graphic HSG and laparoscopic chromopertubation should be employed. The use of color-flow imaging will improve diag­nostic accuracy only in selected cases with an inconclusive spectral Doppler analysis.
Infertility Evaluation and Assisted Reproduction
Summary
Tubal pathology is a major cause of infertility, and so the assessment of tubal patency is an important element in the di­agnostic workup of infertile patients. Conventional methods such as pertubation, radiographic hysterosalpingography, and laparoscopic chromopertubation have disadvantages that in­clude lack of precision, radiation exposure, and invasiveness.
An alternative is the sonographic assessment of tubal patency.
After initial positive experience with transvaginal ultrasound hysterosalpingography following the injection of contrast me­dium, the results could be improved by adding transvaginal pulsed and color duplex sonography to the examination. In our series of 210 infertility patients, tubal patency was evaluated by transvaginal sonography following the intrauterine injec­tion of an absorbable galactose-based contrast medium (Echovist, Schering, Berlin). Depending on the result of the ex­amination (tube patent, partially obstructed, or completely ob­structed), the finding was checked by laparoscopy in 62%, 60 %, and 65% of the cases. The agreement rates between ultrasound and laparoscopy were 95 %, 95%, and 84%, respectively. The contrast medium was very well tolerated. The method pro-
vides an effective, noninvasive screening test for the exclusion of tubal obstruction. The addition of pulsed Doppler ultra­sound provides a level of objectivity that cannot be achieved
with the use of B-mode imaging alone.
References
1 Bald R, Hackelöer BJ: Ultraschalldarstellung verschiedener Endomet-
riumformen. In: Otto R, Jann FX (eds.): Ultraschalldiagnostik. Thieme, Stuttgart 1983
2 Bernaschek G: Vorteile der endosonographischen Diagnostik in Gy-
näkologie und Geburtshilfe. Geburtshilfe Frauenheilkd. 47(1987)471– 476
3 Becker R, Fobbe F, Schlief R, Wolf KJ, Hammerstein J: Prüfung der
Durchgängigkeit der Tubae uterinae durch farbcodierte Duplexsono­graphie mittels eines Ultraschallkontrastmittels (Echovist). Ul­traschall Klin. Prax. Suppl.1 (1988)
3a Campbell S, Bourne TH, Tan SL, Collins WP: Hysterosalpingo contrast
sonography (HyCoSy) and its future role within the investigation of in­fertility in Europe. Ultrasound Obstet. Gynecol. 4 (1994) 245–253
4 Deichert U, van de Sandt M, Lauth G, Daume E: Die transvaginale Hys-
terokontrastsonographie (HKSG) – Ein neues diagnostisches Ver­fahren zur Differenzierung intrauteriner und myometraler Befunde. Geburtshilfe Frauenheilkd. 48 (1988) 835–844
5 Deichert U, Schlief R, van de Sandt M, Juhnke I: Transvaginal hystero-
salpingo-contrast-sonography (Hy-Co-Sy) compared with conven­tional tubal diagnostics. Human Reproduction 4 (1989) 418–424
6 Fitzgerald DE, Drumm JE: Non-invasive measurement of human fetal
circulation using ultrasound: a new method. Brit. Med. J. 2 (1977) 1450–1451
75
Pulsed Doppler and Color Duplex Sonography in the Assessment of Tubal Patency
7 Hata T, Hata K, Senoh D et al.: Transvaginal Doppler Color Flow Map-
ping. Gynecol. Obstet. Invest. 27 (1989) 217–218
8 Hackelöer BJ, Nitschke S, Daume E, Sturm G, Buchholz R: Ultraschall-
darstellung von Ovarveränderungen bei Gonadotropinstimulierung. Geburtshilfe Frauenheilkd. 37 (1977) 185–190
9 Henkel B, Schlief R: Die ambulante Hysterokontrastsonographie
(HKSG) – eine frühzeitige Selektionsmethode in der Diagnostik der mechanisch bedingten Sterilität. Ultraschall Klin. Prax. 1 (1986) 1–11
9a Hüneke B, Lindner Ch, Braendle W: Untersuchung der Tubenpassage
mit der vaginalen gepulsten Kontrastmittel-Doppler-Sonographie. Ultraschall Klin. Prax. 4 (1989) 192–198
9b Kleinkauf-Houcken A, Hüneke B, Lindner Ch, Braendle W: Combining
B-mode ultrasound with pulsed wave Doppler for the assessment of tubal patency. Human Reproduction 12 (1997) 2457–2460
9c Kurjak A: An Atlas of Transvaginal Color Doppler. Parthenon Publish-
ing Group, London 1994
10 Lindner Ch, Braendle W, Schlief R, Bispink L, Luckhardt M, Bettendorf
G: Die sonographische Hysterosalpingographie. Alete Wissen­schaftliche Reihe, 100. Tagung der Nordwestdeutschen Gesellschaft für Gynäkologie und Geburtshilfe (1988) 101–104
7
11 Maathuis JB, Horbach JGM, Van Hall EV: A comparison of the results of
hysterosalpingography and laparoscopy in the diagnosis of fallopian tube dysfunction. Fertil. Steril. 23 (1972) 428–431
12 Nicolini U, Belotti M, Bonazzi B, Zamberletti D, Candiani GB: Can ultra-
sound be used to screen uterine malformations? Fertil. Steril. 47 (1987) 89–93
13 Randolph JR, Ying YK, Maier DB, Schmidt CL, Riddick DH: Comparison
of real-time ultrasonography, hysterosalpingography, and laparas­copy/hysteroscopy in the evaluation of uterine abnormalities and tubal patency. Fertil. Steril. 46 (1986) 828–832
14 Richman TS, Viscomi GN, de Cherney A, Polan ML, Alcebo LO: Fallopian
tubal patency assessed by ultrasound following fluid injection. Radiol­ogy 152 (1984) 507–510
15 Sanfilippo JS, Yussman MA, Smith O: Hysterosalpingography in the
evaluation of infertility: a six-year review. Fertil. Steril. 30 (1978)636– 643
16 Taylor KJW, Burns PN, Wells PNT, Conway DI, Hull MGR: Ultrasound
Doppler flow studies of the ovarian and uterine arteries. Brit. J. Obstet. Gynecol. 92 (1985) 240–246
17 Tristant H, Benmussa M: Atlas der Hysterosalpingographie. Enke,
Stuttgart 1984
76

8 Abnormalities of Corpus luteum Function

S. Kupesic, A. Kurjak, and T. Zodan
Many authors have described the clinical importance of nor­mal corpus luteum function in the initiation of a normal preg-
12
nancy more than two days in the histological development of the en­dometrium relative to the calculated date. It is often a direct re­sult of hormonal dysfunction of the corpus luteum. This dys-
. An inadequate luteal phase is defined as a delay of
function can have a variety of causes, most notably decreased levels of follicle-stimulating hormone (FSH) in the follicular phase, inadequate secretion of luteinizing hormone (LH), decreased LH and FSH levels at the time of ovulation, and poor endometrial responsiveness to progesterone.

Morphology and Biochemistry of the Corpus luteum

Cell types and hormone production. The differentiation of the
corpus luteum is an important event in the ovarian cycle and the critical factor in sustaining an early pregnancy. After ovula­tion has occurred, vessels sprout from the theca to form a net-
work of blood vessels, marking the start of corpus luteum for­mation (Fig. 8.
various cells: K cells in addition to large and small luteal cells.
The large luteal cells develop from the granulosa cells, and the small luteal cells from the thecal cells. The large luteal cells produce more progesterone than the small luteal cells, but the latter appear to be more sensitive to stimulation by LH and chorionic gonadotropin. Also, the small luteal cells appear to produce various angiogenesis factors, and this may occur inde­pendently of the production of steroid hormones. Production of the prostaglandins I cell cultures. The prostaglandins, whose formation is in­fluenced by lipoxygenase products of arachidonic acid such as 5-HETE (hydroxyeicosatetraenoic acid) and is independent of chorionic gonadotropin, act directly on progesterone metabo­lism. Prostaglandin I
1). Ultimately the corpus luteum contains
, and F2αhas been demonstrated in
2,E2
and prostaglandin E2promote pro-
2
gesterone formation, while prostaglandin F action. Besides being regulated by the hypothalamic–pituitary axis (FSH, LH), the corpus luteum has its own paracrine regula­tory mechanism whose details require further investigation.
Luteal phase. The luteal phase begins with the release of the oocyte and the formation of the corpus luteum, accompanied by a significant rise of LH and FSH. The small luteal cells in­creasingly produce LH receptors, which stimulate pro-
gesterone production. The midluteal phase is characterized by peak levels of circulating LH and progesterone and by the lowestresistance index (RI) in the corpus luteum blood vessels, as Kupesic et al. Doppler sonography (Fig. 8. tration finally suppresses the secretion of gonadotropins, the LH and progesterone levels fall, and the RI in the luteal blood
vessels increases. A condition known as luteal phase defect (LPD) can result from faulty “internal” regulation as well as ad-
verse external factors (e.g., strenuous exercise or ovulation­stimulating medications).
has a luteolytic
2α
11
demonstrated by transvaginal pulsed color
2). The rising progesterone concen-
Infertility Evaluation and Assisted Reproduction
Fig. 8.1 Transvaginal ultrasound scan of the ruptured follicle (left).
An increased blood flow velocity and decreased RI (0.44) are typical
signs that indicate ovulation and the start of corpus luteum formation.
Fig. 8.2 Increased blood flow in the mature corpus luteum (left). The Doppler trace indicates a high blood flow velocity and low RI (0.47).
77
Abnormalities of Corpus luteum Function

Conventional Methods in the Diagnosis and Treatment of Luteal Phase Defect

78
Definition. Various terms have been applied to this disorder: luteal phase defect, luteal phase deficiency, short luteal phase, luteal insufficiency, and inadequate luteal phase. All these terms describe the same condition, which consists of a pro­gesterone deficiency, a luteal phase shorter than 11 days, and a delay of 2 or more days in the secretory transformation of the endometrium.
Possible Causes of Luteal Phase Defect
Effect of LH. Zeleznik and Little-Ihrig studied the effect of LH on
corpus luteum function in rhesus monkeys ing hormone (GnRH) was administered to induce gonadotropin secretion. Various plasma LH concentrations were measured, de­pending on the amount of GnRH administered. It was found that an LH concentration of 50 % normal was still able to sustain pro­gesterone secretion during the late luteal phase.
These results confirm the hypothesis that regression of the cor­pus luteum in the nonfertile cycle is due primarily to a reduced
8
luteal cell responsiveness to LH rather than a reduction in gonadotropin secretion.
Jones showed that an imbalance between the FSH and LH levels is responsible for inadequate folliculogenesis and for in­adequate transformation of the granulosa and theca cells into the granulosa luteal cells and theca luteal cells of the corpus lu-
10
teum
. This leads to luteal phase defect. Corpus luteum dys­function with a normal length of the luteal phase may result from impaired granulosa cell function or from an inadequate LH surge with a fairly normal total LH secretion and theca cell response. A short luteal phase is associated with a poor LH surge and low LH secretion. There appears to be a critical LH level that must be maintained after ovulation to ensure the morphological and functional transformation of the granulosa and theca cells and the induction of enzymes for the steroido­genesis of regulatory peptides and various peptide receptors.
Strenuous exercise. Beitinis et al.2studied 28 female students with regular cycles who performed regular, strenuous physical exercise during a two-month training program. All subjects collected daily overnight urine samples for three months: at the start of the study, during a control cycle without exercise, and during two exercise cy­cles. LH, FSH, estriol, and free progesterone were determined and related to creatinine excretion. Twenty cycles with a luteal phase defect were observed in 18 participants. Four women had an inade­quate luteal phase in the first month of the training program, com­bined with decreased free progesterone secretion and a luteal phase shorter than nine days. During the second exercise month,
two inadequate and four short luteal phases were observed. Be­cause disturbances of LH andestriol secretion were also observed in
women with short luteal phases, it may be assumed that regular, strenuous exercise, which was associated with significant weight loss in 12 cases, can indeed lead to anovulation. It is interesting to note, however, that only two of the women had a luteal phase de-
fect in both cycles.
These results show that physical exercise, change of living con­ditions, stress, or other extraneous factors can cause menstrual disturbances that may affect the entire cycle or only the luteal
20
. Gonadotropin-releas-
phase. It appears, however, that these effects are transient in the majority of cases. Clinical practice has shown that many women will show menstrual abnormalities when followed for several months, but that very few of these problems are per­manent.
Ovarian stimulation. There is still disagreement whether ovar­ian stimulation causes luteal insufficiency. Hecht et al. showed that luteal insufficiency is rare in clomiphene-induced cycles. On the other hand, Reshef et al.
15
found that in 30 women who had been treated with gonadotropin and hCG, 27% of the patients showed inadequate endometrial develop­ment.
Diagnosis of Luteal Phase Defect
Endometrial biopsy. One of the greatest problems for scientists
and clinicians is the detection of luteal insufficiency. One op­tion is the histological analysis of endometrial biopsies. This method is fairly precise, since the amount of progesterone pro­duced by the corpus luteum can be estimated by the transfor­mation that the endometrium has undergone for implantation. In a regular cycle, the biopsy is taken shortly before the start of menstrual bleeding on the basis of an assumed cycle length of 28 days and a luteal phase of 14 days duration. Because the luteal phase may last from 12 to 14 days, however, the op­timum timing of the biopsy is disputed. Biopsies that are taken in the early and midluteal phase show greater histological var­iation than biopsies from the late luteal phase. Timing the bi­opsy close to menstruation will best reflect the cumulative progesterone activity.
A major problem is different biopsy interpretations ren­dered by different evaluators or by the same evaluator at different times.
Gibson et al.5obtained duplicate endometrial biopsy samples from 25 women in one sitting. Five colleagues evaluated the two biopsy series on two separate occasions at least two weeks apart. In 43.1% of the cases, the same evaluator gave exactly the same reading for duplicate slides from the same patient, and in 5% of cases the same evaluator saw a difference in endometrial transformation of three or more days in duplicatereadings. Even greater inconsistencies oc­curred among different evaluators. The readings agreed in only 25% of the cases, and in 22 % of cases the discrepancy was greater
than two days.
Serum progesterone level. The serum progesterone level is also used in the diagnosis of luteal insufficiency. Minimum levels of 2.5–5.0 ng/ml in the midluteal phase indicate ovula­tion, while levels of 10–15ng/ml in this phase correspond to normal corpus luteum function tions in progesterone secretion can cause deviations of more than 30 % from the mean value, a single progesterone determi­nation does not reflect either corpus luteum function or its ef­fect on the endometrium.
19
. But because diurnal varia-
8

Ultrasound and Doppler Sonography in the Detection of Luteal Phase Defect

Placental protein. A “placental protein 14” has been described,
which is expressed by the endometrial glands and has occa­sionally been detected in the blood of women with anovula­tory cycles
Dawood
4
. A test procedure has not yet been established.
3
states that the combined use of all test procedures
is necessary in order to draw relevant conclusions.
Treatment of Luteal Phase Defect
Stimulation with clomiphene and hCG. The current treatment
for luteal phase defect is based on optimum follicular develop­ment by stimulation with clomiphene and hCG (human chorionic gonadotropin) or hMG (human menopausal gonadotropin). The response to hCG depends on the age of the corpus luteum. A good increase in progesterone secretion was achieved by administering 5000 IU of hCG 8–12 days after the LH surge. A moderate increase was achieved by administering hCG four days after the LH surge, and no increase was achieved
when it was administered at the time of the LH surge on these observations, the support of corpus luteum function should begin in the midluteal phase, approximately 7–8 days after the LH surge or 6–7 days after the rise in basal body temperature (i.e., on day 21 of a 28-day cycle) domized controlled study, no significant benefit was derived from treatment with progesterone suppositories or oral dehy­drogesterone compared with no treatment. Three comparative studies showed no difference with or without treatment.
19
. Based
3
. In a ran-
FSH. Insler9states that luteal insufficiency is only one of a num- ber of disorders that include an imbalance of intraovarian and extraovarian hormones, peptide interactions, and follicle for­mation at the wrong time of the cycle. Consequently, there are only a small number of women in whom treatment with FSH alone during the early follicular phase can prevent an en­dometrial delay observed in previous cycles from occurring in the next luteal phase.
LUF Syndrome
Another functional disturbance that should be addressed is the syndrome of the luteinized but unruptured follicle (LUF syn­drome). In LUF syndrome, rupture of the preovulatory follicle does not occur despite luteinization. Progesterone production may be deficient in LUF syndrome or may be within normal limits. The postovulatory parameters may also be essentially normal. This can mimic a “presumed ovulatory” cycle, making it very difficult to diagnose LUF syndrome. The syndrome can have various causes: a deficient LH surge, an absence of pre­ovulatory progesterone secretion, primary oocyte abnormali­ties, and changes in progesterone synthesis or other mediators leading to rupture of the follicle. The condition was initially di­agnosed laparoscopically, and later its diagnosis was aided by detecting low concentrations of ovarian steroids in the cul-de­sac fluid. As ultrasound techniques are refined, it is reasonable to expect that new discoveries on this rare syndrome will be forthcoming.
Infertility Evaluation and Assisted Reproduction
Ultrasound and Doppler Sonography in the Detection of Luteal Phase Defect
No diagnostic methods used to date have been sufficiently ac­curate or reliable, making it necessary to introduce new methods into research. Ultrasonography is a highly promising approach, using transvaginal sonography for better identifica­tion of the corpus luteum and combining B-mode and real­time imaging (Fig. 8.
3) with pulsed color Doppler. These tech-
niques are opening up new capabilities for investigations of the corpus luteum, LPD, early pregnancies, and abnormal early pregnancies.
Corpus luteum volume and hormone production. Glock et al.6in-
vestigated whether the size of the corpus luteum as determined by ultrasound or a change in corpus luteum size in early pregnancy correlates with the serum levels of progesterone, E droxyprogesterone or might even serve as a criterion for predicting
the course of the pregnancy. They hypothesized that the corpus lu-
teum volume in early pregnancy correlates with steroid production by the corpus luteum, that the luteal echo pattern based on the relative amount of cystic components correlates with the serum hormone concentration or pregnancy outcome, and that regres­sion of the corpus luteum leads to pregnancy loss. In fact, their study data showed no correlation between changes in corpus lu-
teum volume and changes in steroid production during early preg­nancy. They did find, however, that a decreasing corpus luteum
volume before the 8th week of pregnancy was associated with an increased abortion risk. Color Doppler sonography was used to dis-
tinguish the dominant ovary with the corpus luteum from the con-
tralateral, nondominant ovary (Fig. 8.4). Early pregnancy was
, and 17-hy-
2
characterized by a low RI of 0.39–0.49 in the dominant ovary and a high RI of 0.69–1.0 on the contralateral side. In one patient the RI
was 0.74 in the dominant ovary and 0.79 on the other side. This
high RI on both sides was associated with failure of the pregnancy.
11
Intraovarian RI. Kupesic et al.
47 healthy subjects with ovulatory cycles with that in 28 patients with luteal phase defect (LPD) and 4 patients with LUF syndrome.
They measured the follicular diameter by daily examinations and observed the rupture of the follicle at ovulation as well as the de­marcation of the corpus luteum, presenting as a hypoechoic struc-
ture with an irregular border. The thickened endometrium and the presence of free fluid in the cul-de-sac were simultaneously docu­mented. All of these findings were interpreted as evidence of ovula-
tion. Equivocal cases (in which a corpus luteum was not visualized or follow-ups were not possible) were dropped from the ongoing study. LPD was diagnosed on the basis of progesterone levels and endometrial biopsies in the midluteal phase. The sonographic and Doppler sonographic findings were correlated with measurements of hormone levels and the histopathological data. LUF syndrome
was documented by daily ultrasound examinations and hormone assays. In all four cases of LUF syndrome, normal follicular growth and normal preovulatory follicular diameters were found. The fol­licles maintained their size and shape during the phase in which ovulation was expected to occur. Luteinization of the unruptured
follicle was manifested by increasingly high-level echoes at the
follicular margin.
In the group of subjects with normal ovulatory cycles (n = 47),
different ovarian RI values were measured at different times. Follic-
compared the intraovarian RI in
79
Abnormalities of Corpus luteum Function
Fig. 8.3 Transvaginal ultrasound scan of an ovary that contains a cor­pus luteum. It was not possible with B-mode ultrasound to determine
anything about the functional status of the ovary.
Luteal conversion. Merce et al.14explored all aspects of trans- vaginal sonography—its advantages, disadvantages, current capabilities, and future outlook. In their study on ovarian blood flow during the luteal phase, they introduced the term “luteal conversion” to describe the Doppler phenomena that occur in the luteal phase: conspicuous Doppler signals, an increase in the frequency spectrum, increased blood flow turbulence with a large scatter of maximum frequencies and superimposed waveforms presenting different peak systolic velocities, and fi­nally an increase in the intensity and area of the Doppler sig­nals recorded over the ovary. The same authors in their LPD study observed a fall of the RI in the dominant ovaryduring the luteal phase compared with the follicular phase, like that also occurring in a normal cycle. A significant correlation between RI and progesterone level was not described.
80
8
Fig. 8.4 Color Doppler clearly demonstrates the copious blood
supply to the corpus luteum. The low RI (0.41) is typical of the corpus
luteum.
ular growth and development were associated with moderate to high RI values (0.56 0.06). A significant decrease in RI (p 0.001)
was observed on the day of the LH surge (RI 0.44 ⫾ 0.04). The
lowest RI values were measured in the midluteal phase (RI 0.42
0.06), and these values increased again during the late luteal phase (RI 0.50 0.04). Normal endometrial dating was confirmed by bi­opsy in 15 patients. In the group of patients with LPD (n = 28), no change in ovarian RI was found during the follicular phase (p
0.05). The mean RI throughout the luteal phase was significantly higher (RI 0.56 0.04, p 0.001) than that in the women with nor­mal cycles. No differences were seen between the early, mid- and late luteal phases (p 0.05).
In the control group, the RI in the follicular and luteal phases
was significantly higher on the dominant side (p 0.001). In the LPD group, there were no differences (p 0.05) between the domi­nant and nondominant sides. The mean progesterone level was sig­nificantly lower in the LPD group (p ⬍ 0.001) (6.9 ⫾ 2.3 ng/ml) than in the control group (24.1 ⫾ 11.4 ng/ml). The histopathological
findings showed a delayed endometrial pattern in all of the LPD patients. Progesterone and the RI were correlated in the midluteal phase (r = 0.09, p 0.83). The patients with LUF syndrome (n =4) showed no difference in intraovarian RI values following the LH surge. Similar RI values were measured during the follicular and luteal phases (0.55 0.04 and 0.54 0.06). The mean pro­gesterone level in this group was 14.1 ⫾ 6.2 ng/ml.
LUF syndrome. Merce et al.
14
saw no decrease in the intraovar­ian RI after the LH surge in patients with LUF syndrome. The RI values were in the upper normal range within four days after the LH peak and during the growth and luteinization of the fol­licle. Later they were again similar to those during the follicular phase, indicating that the progression of RI values in LUF syn­drome does not show true cyclic variations, similar to the pat­tern in an anovulatory cycle. “Luteal conversion” does not take place, which shows that the changes in microvascularity de­scribed in normal cycles either do not occur in LUF syndrome or occur in an altered form, possibly due to the failure of follic­ular rupture.
Implantation. Merce et al.
14
also noted the importance of en­dometrial and ovarian blood flow for implantation. They specifically recommended the use of Doppler ultrasound tech­niques in studies that deal with blood flow phenomena during implantation and their relationship to the course of pregnancy.
Intraovarian blood flow and progesterone levels. Glock and Brumsted
values and the progesterone (P) level during the course of the ovar-
ian cycle. The mean P levels in the luteal phase of patients with LPD
were significantly lower than in women with normal cycles (p
0.001). The mean RI in the LPD patients was significantly increased in both the follicular and luteal phase (p= 0.02). The systolic and di­astolic blood flow velocities were decreased in the LPD patients, al-
though these differences were not statistically significant (p = 0.54,
p = 0.11). Correlations between P level and RI were observed in all portions of the luteal phase, thehighest correlation occurring in the midluteal phase (early luteal phase r = 0.73, p = 0.03; midluteal phase r = 0.80, p 0.01; late luteal phase r = 0.63, p = 0.07). The mean RI in the dominant ovary of women with normal cycles was significantly lower throughout the cycle than in the contralateral ovary (0.50 vs. 0.65, p = 0.001). This was not the case in patients
with LPD (0.60 vs. 0.66, p = 0.37). The RI in both ovaries remained elevated in one patient with an anovulatory cycle (mean value 0.76 [0.70–0.82]).
7
studied the correlation between intraovarian blood flow
The study demonstrated a correlation between corpus luteum blood flow and progesterone level during the normal ovarian cycle. The strongest correlation exists in the midluteal phase— the period in which there is maximum neovascularization of the corpus luteum. Consistent with this finding, the authors observed a rise of vascular resistance in the late luteal phase— the period in which the corpus luteum begins to regress. These

Blood Flow in the Corpus luteum during Early Pregnancy

0.64
RI
0.62
0.58
0.54
0.50
0.46
0.42 –8–7 –6 –5 –4 –3 –2 –1 0 1 2 3 4 5 6 7 8 9 10
Days before and after ovulation
LPD patients Control group
Fig. 8.5 Changes in intraovarian blood flow before and after ovula-
tion in luteal phase defect (LPD) patients and a control group.
findings suggest that measuring the RI in the corpus luteum may be a useful adjunct to the plasma progesterone assay in the evaluation of luteal function.
Tinkanen et al.
18
, on the other hand, found no differences in luteal blood flow measurements between healthy subjects and infertile patients. The authors found that a shortened luteal phase was not associated with a premature regression of cor­pus luteum vascularization.
Strigini et al.
17
investigated the changes of vascular re­sistance in patients with FSH-treated cycles. The uterine pul­satility index (PI) in stimulated cycles was significantly lower than in normal cycles both before and after ovulation. This was attributed to an increased E
Doppler findings, endometrial biopsy, and hormone levels.
Kupesic et al.
12
correlated Doppler flow measurements with histo-
concentration in the plasma.
2
logical findings and hormone studies.They presumed that the diag­nosis of a luteal phase defect could be improved by combining ul-
trasound results with endometrial biopsy and hormonal markers. The mean progesterone levels in women with LPD were signifi-
cantly lower than in the control group (10.2 4.3 ng/ml vs. 21.0
4.2 ng/ml, p 0.01). The FSH/LH ratio was significantly lower in the group with a delayed endometrial pattern (p ⬍ 0.001). A close cor- relation was found between estradiol levels and follicular size on days – 5 to – 1. An increase in follicular diameter and endometrial
thickness was observed in both groups.
No differences in intraovarian flow resistance were found in the
proliferative phase (p 0.05). The control group showed a signifi-
0.72
RI
0.70
0.68
0.66
0.64
0.62
0.60
0.58
0.56
0.54
0.52
0.50
0.48 –8–7 –6 –5 –4 –3 –2 –1 0 1 2 3 4 5 6 7 8 9 10
Days before and after ovulation
LPD patients Control group
Fig. 8.6 Blood flow changes in the spiral arteries before and after ovulation in luteal phase defect (LPD) patients and a control group.
cant regression of the RI on the day of the LH surge (RI = 0.45
0.04, p 0.05). Thereafter the RI again rose to the values that were measured during the follicular phase (RI = 0.49 ⫾ 0.02). The mean RI was significantly higher in the LPD group (RI = 0.58 ⫾ 0.04, p
0.001) (Fig. 8.5) than in the control group. The RI in the dominant ovary of the control group was significantly lower than in the con-
tralateral ovary, whereas both sides were equal in most of the LPD patients. Blood flow in the spiral arteries was also measured. The RI in the control group was 0.53 0.04 in the periovulatory phase,
0.50 0.02 in the midluteal phase, and 0.51 0.04 in the late luteal phase (Fig. 8.6). In the LPD group, increased resistance values
were measured in the spiral arteries duringall phases (periovulatory phase RI =0.70 0.06, p 0.001; midluteal phase RI= 0.72 0.06,
p ⬍ 0.001; late luteal phase RI = 0.72 0.04, p ⬍ 0.001). A close correlation was found between plasma estradiol levels and follicular diameter.
The study shows that, in women with normal endometrial development, a decline of vascular resistance occurs in the uterine, radial, and spiral arteries from the follicular phase to the luteal phase. On the other hand, retarded endometrial development is marked by a rise of vascular resistance in the uterine vessels during the luteal phase. Since the most signifi­cant differences were found in measurements of the spiral ar­teries, the changes in endometrial blood flow could provide an important parameter for predicting endometrial development and the likelihood of implantation.
Infertility Evaluation and Assisted Reproduction
Blood Flow in the Corpus luteum during Early Pregnancy
Normal and abnormal early pregnancy. Salim et al.16compared
corpus luteum blood flow in normal and abnormal early pregnan­cies. Their study tested the hypothesis that the absence of meas­urable luteal blood flow did not correlate with a normal early preg­nancy. The resistance in the intraovarian vessels in women with an abnormal early pregnancy (missed abortion, incomplete or
threatened abortion) was significantly higher (p 0.01) than in women with a normal early pregnancy. This did not apply to
patients with a molar or ectopic pregnancy, however.
The differences between the subgroups of abnormal early pregnancy may be due to the different nature of the causes. Missed abortion and incomplete abortion present as an early loss of pregnancy with no potential for further development.
Threatened abortion is a similar situation. It is unclear whether decreased blood flow in the corpus luteum is a potential cause of this condition or a result. Molar and ectopic pregnancies present a different situation. In these cases the pathological
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