Heterotopic triplet pregnancy: report of a case with bilateral tubal pregnancy and an intrauterine pregnancy

The first report of an ectopic pregnancy following IVF was published in 1976, and since then heterotopic pregnancies (HPs) have been reported at an increasing rate. Although cases of the co-existence of a bilateral tubal and an intrauterine pregnancy following IVF-embryo transfer have been reported, a case of heterotopic triplet pregnancy caused by unilateral tubal embryo transfer has not yet been published in the literature. Here we report on a 38-year-old women (gravida 3, para 1) with a history of infertility who presented to our infertility clinic for evaluation. Hysterosalpingography revealed bilaterally patent Fallopian tubes and stricture of the cervical canal. She conceived after receiving HMG combined with pure FSH, followed by IVF-tubal embryo transfer. Four embryos were replaced into the right tube. Approximately 5 weeks after tubal embryo transfer, the patient presented with lower abdominal tenderness and shock due to internal bleeding. She underwent an emergency laparotomy under the impression of HP. Bilateral tubal pregnancy with right tubal rupture was noted during the operation. The post-operative course was uneventful. Early intervention and thorough inspection of the peritoneal cavity in patients with haemodynamic instability can prevent jeopardizing the life of the mother as well as the ongoing pregnancy.     

Analysis of the incidence and risk factors associated with ectopic pregnancy following in-vitro fertilization and embryo transfer

Ectopic pregnancy is a well known complication of in-vitro fertilization(IVF) and embryo transfer. From March 1983 to December 1993,3000 clinical pregnancies were achieved at Bourn Hall Clinic,including 135 ectopic pregnancies (4.5%). Of these ectopics20 were heterotopic, eight ovarian, six bilateral tubal andthe remainder were singleton tubal pregnancies. The main riskfactor identified in the series was a history of pelvic inflammatorydisease (P < 0.001). The data also showed that ectopic pregnancyis at present more prevalent among patients in whom tubal damageis the reason for treatment. There was slight statistical evidence(P = 0.05) that patients having ectopic pregnancies receiveda higher volume of culture medium than those having normal deliveries.There was also an apparent trend (P = 0.07, not significant)that high progesterone/oestradiol ratio on the day of embryotransfer was associated with ectopic pregnancy. There was nostatistical evidence of association between ectopic pregnancyand a history of ectopic pregnancy, abortion, still birth, terminationof pregnancy, neonatal death, tubal surgery, ovarian stimulationprotocol, plasma concentration of oestradiol, luteinizing hormoneand progesterone, number of oocytes retrieved, number or qualityof embryos transferred, administration of general anaesthesiafor embryo transfer, and the number of patent Fallopian tubes.Awareness of the risk factors associated with ectopic pregnancyplays an important part in the early diagnosis of this potentiallyfatal condition.     

Rare ectopic pregnancies after in-vitro fertilization: one unilateral twin and four bilateral tubal pregnancies

Between 1985 and 1989, one unilateral twin and four bilateral tubal pregnancies were encountered among 124 extrauterine pregnancies and 1648 intrauterine pregnancies following in-vitro fertilization and embryo transfer. The two factors associated with this high incidence of single and multiple extrauterine pregnancies were tubal damage and multiple embryo transfer. Embryos at different stages of development appear to have the capacity to implant ectopically. Despite advances in diagnostic capabilities, ectopic pregnancy remains a major cause of maternal mortality. Early diagnosis prior to rupture must be made if mortality and morbidity are to be abolished. The use of transvaginal sonography has improved the diagnosis of ectopic pregnancy and should be routinely used in all pregnancies following assisted conception. The identification of an intrauterine pregnancy should not be sufficient to rule out the possibility of an extrauterine pregnancy or even bilateral tubal pregnancies.     

Bilateral tubal ectopic pregnancies: a report of two cases

Bilateral tubal ectopic pregnancies are rare occurrences. Two recently managed cases are discussed. The first was a single, sexually active 23-year-old nullipara with family history of twinning who presented with eight weeks amenorrhea, positive pregnancy test, lower abdominal discomfort and other clinical and ultrasound findings suggestive of unruptured left tubal pregnancy. Intra-operatively, unruptured bilateral tubal pregnancies were found and bilateral salpingotomy performed with uneventful recovery. Histology of the specimens confirmed the intra-operative diagnosis. She was appropriately counseled. Case 2, a 30-year-old multiparous housewife who had been on clomid for secondary infertility, presented with signs and symptoms of ruptured tubal ectopic. Intra-operatively, ruptured left and unruptured right tubal pregnancies were found and salpingectomy and salpingotomy were done respectively, with uneventful recovery. The diagnosis was also confirmed histologically and counseling given as in case 1. Bilateral tubal ectopic pregnancies appear to be increasing with twin proneness and use of fertility drugs as risk factors. Whether spontaneous or induced, the hallmarks of good management include early presentation, high index of suspicion, meticulous ultrasound scanning, good case selection, judicious intra-operative inspection of the contralateral tube, histology of specimens and appropriate patient counseling.     

Analysis of ectopic pregnancies resulting from in-vitro fertilization and embryo transfer

The purpose of this study was to analyse the risk factors, stimulation characteristics, site and outcome of pregnancy and future fecundity of patients who develop ectopic pregnancies after in-vitro fertilization (IVF). Of 3145 transfer cycles between January 1981 and July 1989, 27 (3.3%) of the resulting 825 pregnancies were ectopic. There was a significantly greater incidence of a prior ectopic pregnancy in the study group compared to the controls. Compared to matched controls with intrauterine pregnancies, the study group had significantly higher peak oestradiol levels. Twenty-one ectopic pregnancies were ampullary, two were interstitial, one was abdominal, one was cervical and two were heterotopic. Sixteen of the patients subsequently underwent 40 IVF attempts with a pregnancy rate of 28% per transfer. We conclude that patients with a prior ectopic pregnancy are at risk for an IVF ectopic pregnancy. The subsequent IVF outcome of those who develop ectopic pregnancies after IVF is encouraging.     

Ectopic pregnancy after tubal sterilization

We report 13 cases of ectopic pregnancy following tubal ligation out of 287 ectopic pregnancies seen during a six year period (1984-1989). These findings suggest that tubal sterilization does not invariably confer infertility. Ectopic pregnancy must not be disregarded in women who have undergone tubal ligation, especially if two or more years have elapsed since the sterilization.     

Modelling of the probability of success of the stages of in-vitro fertilization and embryo transfer: stimulation, fertilization and implantation

The aim of this study was to find the factors explaining the probability of success of in-vitro fertilization (IVF)-embryo transfer and its different stages: stimulation, fertilization and implantation. The sample came from a retrospective cohort followed in the IVF-embryo transfer centre of the Centre Hospitalier Universitaire of Pellegrin, Bordeaux, France; the data from 471 couples giving rise to 923 IVF- embryo transfer cycles were recorded. Four logistic regression models were specified for global process, stimulation, fertilization and implantation stages. Random effect models were used for taking into account the correlations of the different cycles for the same woman. The main outcome measures were: ongoing pregnancy, number of oocytes, number of embryos. A total of 135 ongoing pregnancies was observed. The significant explanatory variables were for global process: age [> or = 38 years: odds ratio (OR) = 0.28], donor sperm IVF-embryo transfer (OR = 2.1), number of ampoules (OR = 0.98), previous IVF-embryo transfer livebirth (OR = 2.36); for stimulation: age (> or = 35 years: OR = 0.38) and number of days of treatment > 13 days (OR = 0.20); for fertilization: accident during previous IVF-embryo transfer gestation (OR = 0.39), absolute tubal infertility (OR = 1.38) and the number of ampoules of human menopausal gonadotrophin (HMG) per day (OR = 0.85); for implantation: age (age > or = 38 years: OR = 0.34), donor sperm IVF- embryo transfer (OR = 4.58), number of ampoules (OR = 0.98), number and quality of the embryos. Estimates of the probabilities of success are also given for the global process.      

Pregnancy: Analysis of the risk factors with regard to the occurrence of ectopic pregnancy after medically assisted procreation

In a retrospective study (1985–1989) based on data fromthe Centre for Reproductive Medicine in Brussels, a total of23 ectopic pregnancies (2.24%) occurred after 3800 embryo, zygoteor gamete transfers. This number was low compared with the datapublished elsewhere. Tubal damage was a major risk factor towardsdeveloping an ectopic pregnancy after in-vitro fertilizationand embryo transfer. The number of ectopic pregnancies afterthe association of clomiphene citrate and human menopausal gonadotrophin(HMG) was significantly higher in patients with tubal (7.8%)and non-tubal indications (2.1%) compared with those stimulatedwith gonadotrophin-releasing hormone (GnRH) and HMG (2.18% and0.84%, respectively). The number of replaced embryos was notassociated with the rate of ectopic pregnancy and neither didtransfer technique (intra-uterine or intra-Fallopian transfer)influence the ectopic pregnancy rate.     

Pregnancy: The prediction of ectopic pregnancy after in-vitro fertilization and embryo transfer

Data from 135 patients who suffered ectopic pregnancies andfrom 135 patients who progressed to singleton deliveries afterin-vitro fertilization and embryo transfer have been analysedretrospectively. The ectopic pregnancies represent all suchcases observed at Bourn Hall Clinic between 1983 and 1993. Thedelivered group was randomly selected from the same time period.The ectopic pregnancies included 20 heterotopic, eight ovarianand six bilateral tubal pregnancies; the remainder were singletontubal pregnancies. The aim of this study was to identify thevariables which differed systematically for the two groups ofpatients and to explore whether such variables could be usedto predict ectopic pregnancy at an early stage. The mean plasmaconcentration of human chorionic gonadotrophin and progesteronefor the ectopic pregnancy group was significantly lower thanthat for the singleton delivery group (P < 0.001). However,there was such a degree of overlap that it was impossible todevise a cut-off concentration for either hormone which wouldoffer a clinically useful predictor of ectopic pregnancy. Nevertheless,using the discriminant function analysis of these data, togetherwith the history of pelvic inflammatory disease, we could predictup to 90% of cases of ectopic pregnancy by day 23 after embryotransfer, long before ultrasound imaging would be useful.     

Aberrant alteration of vascular endothelial growth factor-family signaling in human tubal ectopic pregnancy: what is known and unknown?

More than 98% of ectopic pregnancies occur in the Fallopian tube. Because many facets of tubal ectopic pregnancy remain unclear, prediction, prevention and treatment of tubal ectopic pregnancy are still a major clinical challenge. Compelling evidence suggests that angiogenic growth factors are involved in normal and abnormal implantation. While acknowledging the importance of an intrauterine pregnancy requires the development of a local blood supply and angiogenesis, we hypothesize that the hypoxic- and estrogen-dependent regulation of vascular endothelial growth factor/placental growth factor expression, secretion, and signaling pathways that are possibly involved in the pathophysiology of tubal ectopic pregnancy. Our hypothesis may also lead to a new therapeutic strategy for women with tubal ectopic pregnancy.     

An unexpected triplet heterotopic pregnancy after replacement of two embryos

We report a case of a triplet heterotopic pregnancy consisting of an intrauterine monozygous twin pregnancy and a tubal pregnancy after replacement of only two embryos in an in-vitro fertilization cycle with donor spermatozoa. This case demonstrates that sonographic demonstration of two intrauterine pregnancies after transfer of two embryos does not exclude the presence of an ectopic pregnancy. As both heterotopic pregnancy and spontaneous monozygotic twinning are more frequent after the use of assisted reproductive techniques, this combination, although extremely rare, must be kept in mind, especially in older patients with pre-existing tubal damage.      

Apoptosis versus necrosis in tubal ectopic pregnancies following Methotrexate

Methotrexate administration for the treatment of tubal ectopic pregnancies has been shown to cause tubal mass enlargement. Our hypothesis was that, by administrating Methotrexate, a local necrotic reaction occurs, leading to hematoma formation and eventually fallopian tube rupture. Salpingectomy specimens were collected, analysed and divided into three equal groups: patients who received Methotrexate but who ultimately failed medical treatment, patients who had a viable ectopic pregnancy and patients with a self-resolving ectopic pregnancy that were operated due to other medical indications. The specimens were dyed using the Cleaved Caspase-3 (Asp175) Rabbit mA. Specimens were divided into three equal groups and analysed. The patients in self-resolving ectopic pregnancy group were older and had more pregnancies. Rates of apoptosis were found to be less than 1% per slide. Necrosis was not evident in any of the pathological specimens. It seems Methotrexate administration does not lead to a significant tubal necrotic reaction. Further studies are required.     

Preliminary results on the role of embryonic human chorionic gonadotrophin in corpus luteum rescue during early pregnancy and the relationship to abortion and ectopic pregnancy.

The precise mechanisms by which corpus luteum (CL) function is modulated during early pregnancy are not known. Evidence in failed pregnancies (ectopic, abortions), shows that factors other than human chorionic gonadotrophin (HCG) could be involved in its regulation. The objective of this study was to investigate the dynamics of beta-HCG, progesterone and oestradiol production in early pregnancy and its relation to embryonic quality and topographic localization. Plasma concentrations of progesterone, oestradiol and beta-HCG were studied between days +12 and +21 after an in-vitro fertilization (IVF) embryo transfer in 11 intrauterine pregnancies, 10 intrauterine abortions and seven tubal pregnancies. Tubal pregnancies and abortions were grouped according to doubling time (DT) of HCG. Results showed that oestradiol concentrations were apparently reduced in both ectopic pregnancies and abortions compared with normal pregnancies. The fall in oestradiol concentrations was seen in ectopic pregnancies with an abnormal DT for HCG and in all abortions. When the ectopic pregnancy had a normal DT, oestradiol and progesterone concentrations were normal. In abortions, the fall in oestradiol and progesterone concentrations was less influenced by the DT of HCG. These findings suggest that corpus luteum function depends on an adequate DT of HCG more than an absolute value, and with normal trophoblastic tissue the site of implantation does not affect CL function.     

Serum HCG 12 days after embryo transfer in predicting pregnancy outcome

BACKGROUND: Assisted reproduction treatment (ART) entails a risk of ectopic pregnancy and early pregnancy loss. Serum HCG has been found to be predictive of pregnancy outcome. Our aim was to assess the clinical value of a single early HCG assay in ART pregnancies taking into account the aetiology and treatment of infertility. METHODS: During 1994-1999, we studied 774 embryo transfer cycles resulting in pregnancy defined as a serum HCG concentration of > or =5 IU/l on day 12 following embryo transfer. The treatment included IVF in 518, ICSI in 119, and frozen embryo transfer in 137 cycles. Serum HCG concentrations were measured by fluoroimmunometric assay. Pregnancies were classified as viable (live fetus at > or =22 weeks gestation) or non-viable (biochemical pregnancy, miscarriage, ectopic pregnancy and molar pregnancy). Data on the outcomes were retrospectively retrieved from the records. RESULTS: The median HCG concentration was 126 IU/l in viable pregnancies and 31 IU/l in non-viable pregnancies (P < 0.0001). The median HCG concentration was 115 IU/l in singleton pregnancies and 201 IU/l in multiple pregnancies (P < 0.0001). Male factor infertility was associated with viable pregnancies (P = 0.004) and tubal factor with non-viable pregnancies (P = 0.003); the lowest HCG level (88 IU/l) was observed in subjects with both male factor infertility and ICSI treatment (P = 0.001). An HCG value of 76 IU/l emerged as the most suitable cut-off point to predict viable pregnancy. Probabilities of each type of outcome related to the HCG level are given. CONCLUSIONS: A single HCG reading on day 12 after embryo transfer helps to plan the subsequent follow-up. Male factor infertility and ICSI are associated with relatively low HCG values in viable pregnancies.     

Spontaneous interstitial pregnancy on a tubal stump after unilateral adenectomy followed by transvaginal colour Doppler ultrasound

Transvaginal colour and angio Doppler blood flow analysis combined with serial measurement of human chorionic gonadotrophin (HCG) concentration is reported here for the first time to study the local vascularity of a cornual pregnancy and to monitor the effectiveness of medical therapy. Interestingly, a strong relationship between morphological changes of trophoblastic tissue and the intensity of neovascularization was noted. Systemic single-dose methotrexate (MTX) therapy allowed successful treatment of an interstitial ectopic pregnancy involving part of the proximal portion of a tubal stump. We suggest that, by adding colour Doppler to conventional transvaginal ultrasonography, the outpatient surveillance of ectopic pregnancy evolution following MTX therapy is greatly enhanced. This is of particular value in cornual pregnancies which are highly likely to develop harmful complications during surgical intervention or even during puncture for local MTX injection.      

Bcl-2 expression as a novel immunohistochemical marker for ruptured tubal ectopic pregnancy

Programmed cell death by apoptosis occurs in fetal and maternal tissues during early pregnancy and plays an important role during implantation, decidualization, and in fetal development. In the regulation of apoptosis, bcl-2 is one of the central controlling genes, and acts by protecting the cell against apoptosis. It is postulated that invasiveness of ectopic trophoblast towards and through the muscularis zone of the tubal wall consequently leading to tubal rupture might be due to disturbed regulation of apoptosis. By means of immunohistochemistry and a computerized image analysis, bcl-2 immunostaining was localized and quantified in 36 randomly selected paraffin-embedded ectopic trophoblast tissue specimens collected from women undergoing surgery for ruptured (n = 18) and non-ruptured (n = 18) tubal ectopic pregnancies. Immunostaining was found in the villi syncytiotrophoblast in all patients, while the percentage of positive bcl-2 immunostained area (%PA) (P = 0.0009) and staining intensity (P = 0.0042) were consistently greater in the group of ruptured ectopic pregnancies. Including the variables %PA and saturation into a logistic regression model for a probability threshold of 0.5 (<0.5 = non-ruptured ectopic pregnancy, >0.5 = ruptured ectopic pregnancy) to identify tubal rupture, a sensitivity and specificity of 94.4% were found. It is suggested that elevated bcl-2 immunostaining in the syncytiotrophoblast layer reflects unlimited cell survival of ectopic trophoblast and could lead to the establishment of a circulating marker for tubal rupture.     

Leukemia inhibitory factor (LIF) is immunohistochemically localized in tubal ectopic pregnancy

Leukemia inhibitory factor (LIF) is essential for implantation of the embryo in the endometrium. It is not clear whether the blastocyst requires expression of LIF for implantation into tissues other than endometrium. Immunohistochemical localization of LIF was performed in the fallopian tube of 20 women with ectopic pregnancies, 7 women with normal pregnancies and 20 healthy non-pregnant women. Fallopian tubes were evaluated from specimens taken during tubal ligation in normal pregnancies and non-pregnant fertile women or at operation for tubal surgery in ectopic pregnancies. Biopsies were assayed by immunohistochemistry. Semi-quantitative immunohistochemical reaction scores (IRS) were used for immunohistochemical analyses. Immunolabeling of LIF was detected in the surface epithelium and stroma of fallopian tubes in all subjects. IRS score in the epithelium and stroma of non-pregnant women and women with intrauterine pregnancy were similar (p>0.05). However, women with ectopic pregnancy had significantly increased labeling of LIF compared to others (p<0.05). Immunohistochemical labeling of LIF in the fallopian tube was found to be increased in ectopic pregnancies compared to non-pregnant and healthy pregnant controls. This may indicate a role of LIF in the ectopic implantation of embryos.     

Salpingectomy or proximal tubal occlusion of unilateral hydrosalpinx increases the potential for spontaneous pregnancy

BACKGROUND: Studies carried out over the past 10 years have suggested that hydrosalpinges reduce the pregnancy rate in IVF. Here we report our observations of spontaneous pregnancies in patients who underwent salpingectomy (n = 18) or proximal tubal occlusion (n = 7) following diagnoses of unilateral hydrosalpinges and patent contralateral tubes. METHODS: This multi-centre, retrospective study included 25 infertility patients with known unilateral hydrosalpinges with a patent contralateral Fallopian tube. Laparoscopic treatment of unilateral hydrosalpinges by either salpingectomy or tubal occlusion was performed in each patient. Rates of subsequently observed spontaneous pregnancy, and time to pregnancy, are reported. RESULTS: The average duration of infertility in these patients was 3 years with a range of 1-10 years. Following laparoscopic surgical treatment, a total of 22 patients (88%) achieved intrauterine pregnancies, all without IVF treatment. Pregnancies occurred in an average of 5.6 months with a range of 1-21 months. There were no ectopic pregnancies in the study population. CONCLUSIONS: Selected patients with unilateral hydrosalpinges and a patent contralateral Fallopian tube may exhibit increased cycle fecundity after salpingectomy or proximal tubal occlusion of the affected tube and conceive without the need for IVF.     

Occurrence of ectopic pregnancy among women with recurrent spontaneous abortion

Ectopic pregnancy is a major health problem accounting for about 10% of all maternal mortality. To determine whether patients with a history of recurrent spontaneous abortion (RSA) have a higher frequency of ectopic pregnancy than the general population, the obstetrical histories of 630 patients with a diagnosis of RSA from two centers were studied. Thirty-eight of 376 USA women, (10%) and 10 of 193 British women (5%) previously had suffered an ectopic pregnancy. These data provide an overall prevalence of 9% for ectopic pregnancy among patients who recurrently abort. The total number of pregnancies was 1,716 in the USA group and in the British group was 1,270, for a total of 2,986 including live births, stillbirths, abortions, hydatidiform moles, and ectopic pregnancies. In the US series, 45 of the 1,716 pregnancies were ectopic. If these women had the same relative proportion of ectopic pregnancies as the general population, there should have been 20.6 ectopic pregnancies. This represents a 2.2-fold increase in patients with a history of RSA. There was no association between pregnancy order and occurrence of ectopic pregnancy. The prevalence of known risk factors for ectopic pregnancy in patients with a history of ectopic pregnancy was PID, 3%; tubal surgery, 20%; and used IUD, 3%. Seventy-four percent of the women had no identifiable risk factor. These results suggest a comorbidity for ectopic pregnancy and RSA which may indicate shared etiologies.     

Immunolocalization of integrins and fibronectin in tubal pregnancy

Integrins are a large family of cell adhesion molecules that serve as receptors involved in cell-to-cell and cell-to-matrix interactions during implantation. We studied immunohistochemical staining of integrins (alpha 3, alpha V, beta 1, and alpha 2 beta 1) and fibronectin in ectopic tubal pregnancy. Thirty fallopian tube samples with ectopic pregnancies and five normal tubal segments were obtained during ligation operations; the latter specimens served as controls in the study. Formalin-fixed paraffin-embedded tissue sections were stained with hematoxylin-eosin or primary antibodies against alpha 3, beta 1, alpha V, and alpha 2 beta 1 integrins and fibronectin, using the avidin-biotin-peroxidase method. A semi-quantitative grading system was used to compare staining intensities. In the control samples, immunostaining of all integrins was found in a single layer of tall columnar epithelial cells, the lamina propria (Lp) and the muscular layer. Fibronectin staining was detected in the Lp and the muscular layer. Staining intensities of alpha 3 and beta 1 integrins and fibronectin were increased in the normal part of fallopian tubes with ectopic pregnancies. Staining of beta 1 integrin was more intense than staining of alpha 3 and fibronectin, whereas there was no difference in alpha V and alpha 2 beta 1 integrin expression between normal tubal tissue in the ectopic pregnancy group and control tubal tissue. In the tubal pregnancy group at the site of implantation, staining intensity of alpha 3 and beta 1 integrins and fibronectin was strong in decidual cells, supporting tissue and placental villi, whereas alpha V and alpha 2 beta 1 staining was mild. We concluded that integrins, especially beta 1 and alpha 3, and fibronectin may play a role in progression of tubal implantation. Although the role of integrins has not yet been clearly defined, these molecules may function as markers of normal and abnormal states of receptivity. We like to suggest that integrins and fibronectin, which are needed in utero implantation, are expressed in tubal tissues during ectopic pregnancy and are involved in ectopic implantation.