Ectopic pregnancy in a non-patent fallopian tube following transfer of embryos to the contralateral tube.

A case is reported of ectopic pregnancy in the non-patent leftFallopian tube after transfer of two pronuclear-stage embryosinto the right tube following in-vitro fertilization.     

Effect of mifepristone on the expression of cytokines in the human Fallopian tube

Cytokines are believed to play a critical role as mediators between the oviduct and the developing embryo. A synchronous development of embryo and endometrium is essential to successful implantation. It seems to be beneficial for embryo development to rest for some time in the Fallopian tube. Expression of cytokines in the human Fallopian tube and the effect of mifepristone were investigated. Fourteen women with regular menstrual cycles and proven fertility, admitted to the hospital for tubal ligation, were randomly allocated to control or treatment groups. Mifepristone 200 mg was given on day LH+2. Surgery was performed on day LH+3 to LH+5. Biopsies were obtained from the ampullar and isthmic regions of the tubes. Expression of interleukin 8 (IL-8), tumour necrosis factor alpha (TNFalpha), transforming growth factor beta (TGFbeta) and leukaemia inhibitory factor (LIF) was analysed using immunohistochemistry. All cytokines except IL-8 showed the same staining intensity both in the ampullar and isthmic region, while IL-8 was more pronounced in the ampullar region in both epithelial and stromal cells. Exposure to mifepristone made the spatial difference in IL-8 disappear and increased the expression of TNFalpha in the epithelium of the isthmus, but had no effect on the expression of TGFbeta1 or LIF. Changes in cytokine expression in the Fallopian tube are likely to influence embryo development, which could contribute to the contraceptive effect of mifepristone.     

Differential impact on pregnancy rate of selective salpingography, tubal catheterization and wire-guide recanalization in the treatment of proximal Fallopian tube obstruction

A total of 66 patients with proximal Fallopian tube (113 tubes)obstruction, as diagnosed by both laparoscopy and hysterosalpingogram,were each subjected to a transcervical recanalization proceduresequentially using selective salpingography followed, if necessary,by tubal catheterization with a soft Teflon 2-French catheterand finally, if needed, wire-guide cannulation. Each procedurewas terminated once patency had been achieved without recourseto the next technique. Bilateral obstruction was present in47 patients and unilateral in 19 patients. Patency was achievedin 39 (34.5%) Fallopian tubes by selective salpingography alone,in 52 (46.0%) by tubal catheterization and in 10 (8.9%) by wireguide, with 12 (10.6%) tubes remaining obstructed. Pregnancyoccurred in 24 (36.4%) patients without recourse to other treatment(mean follow-up, 17 months). Where patency was achieved (59patients), 19 out of 43 (44.1%) of those treated for bilateralobstruction and five out of 16 (31.3%) of those treated forunilateral obstruction achieved a pregnancy. Pregnancy occurredin six out of 22 patients (27.3%) where selective salpingographywas used to produce tubal patency, in 17 out of 30 patients(56.7%) where tubal catheterization was used and in one outof seven (14.3%) where a wire guide was used, which was an ectopicpregnancy. The difference between the ongoing pregnancy ratesfollowing tubal catheterization (50.0%) and wire-guide cannulation(0.0%) was significant (P = 0.033). While wire-guide cannulationis the most effective method used to achieve tubal patency,these results indicate that when it is truly necessary, as opposedto electively used by clinicians, the prognosis with regardto pregnancy is poor and alternative therapy such as microsurgeryor in-vitro fertilization should be considered early.     

Leukaemia inhibitory factor receptor and gp130 in the human Fallopian tube and endometrium before and after mifepristone treatment and in the human preimplantation embryo

Leukaemia inhibitory factor (LIF) is a cytokine, which is associated with reproductive processes such as embryo development and implantation. The objectives of this study were to detect the presence of LIF receptor (LIFR) and glycoprotein 130 (gp 130) in the human Fallopian tube, endometrium and preimplantation embryo and to study the effect of mifepristone on the expression of LIFR and gp130 in the Fallopian tube. Twenty-two healthy fertile women received a single dose of 200 mg mifepristone or placebo immediately after ovulation (LH + 2). Biopsies were obtained from the Fallopian tubes during laparoscopic sterilization once between days LH + 4 and LH + 6 and from endometrium once between days LH + 6 and LH + 8. Preimplantation embryos were received from couples undergoing in vitro fertilization treatment. Immunohistochemistry was used to detect the presence of LIFR and gp130 in the Fallopian tube, endometrium and preimplantation embryo. Real-time PCR was used to study LIFR and gp130 expression in the Fallopian tube and endometrium. LIFR and gp130 were localized in the Fallopian tube, preimplantation embryo and endometrium. LIFR was more abundant in the Fallopian tube than in the endometrium. In the blastocyst, the staining of gp130 was mainly localized in the inner cell mass, whereas LIFR was expressed in all cells. The presence of LIFR and gp130 in the Fallopian tube and preimplantation embryo indicates a role for LIF in communication between the embryo and the Fallopian tube. Mifepristone did not affect the expression of LIFR and gp130 in the Fallopian tube, nor in the endometrium suggesting that progesterone might not be directly involved in the regulation of LIFR or gp130.     

Molecular aspects of implantation: Integrins {beta}5, {beta}3 and {alpha}v are apically distributed in endometrial epithelium

Several adhesion molecules have been shown to occur at the surfaceof endometrial cells. One of these is the integrin     

Prostaglandin E2 and F2alpha receptors in the human Fallopian tube before and after mifepristone treatment

Prostaglandins are associated with several reproductive processesin addition to their effects on the vascular system and muscularcontractility. The aim of this study was to gain informationabout the localization of the receptors for PGE₂ (EP1–EP4)and PGF₂ (FP) in the human Fallopian tube and their regulationfollowing treatment with mifepristone. Sixteen healthy fertilewomen received a single dose of 200 mg mifepristone or placeboimmediately after ovulation (LH+2). Laparoscopic sterilizationwas performed on days LH+4 to LH+6. Biopsies were taken fromthe Fallopian tubes bilaterally. The expression of EP1, EP2,EP3, EP4 and FP was analysed using immunohistochemistry andRT–PCR. The co-localization of prostaglandin receptorsand c-kit or e-nos was analysed using confocal microscopy. Theeffect of progesterone, mifepristone and prostaglandin on tubalcontractility was studied. The presence of EP1–EP4 andFP in the Fallopian tube was detected using immunostaining.The receptors were expressed in serosal cells, luminal epithelialcells, and the muscular wall and vessels of the Fallopian tube.Co-localization studies showed that the endothelial cells stainedpositive for EP1–EP4 and FP and that co-localization wasseen for EP4 and c-kit. Decreased contractility was seen afterprogesterone treatment, whereas increased contractility wasseen after PGF₂ and PGE₂ treatment. These data suggest thatboth the transport of the embryo and the communication betweenthe embryo and the Fallopian tube involve the action of prostaglandinsthrough EP and FP receptors in addition to the effect of prostaglandinson the vascular system and muscular contractility.     

Immunology: Evidence for the in-vitro de-novo synthesis of immunoglobulin and a previously undescribed 17 kDa protein (TEP-2) by the mucosa of the Fallopian tube

De-novo synthesis and secretion of proteins by short-term explantsof matched Fallopian tube mucosa and endometrium were studiedusing radiolabelled L-[³⁵S]methionine and [³H]glucosamine. Tocompare directly each anatomical site of the Fallopian tubeand endometrium from the same source, newly synthesized proteinswere separated on one-dimensional polyacrylamide gel electrophoresisand examined by autoradiography. De-novo synthesis of two proteinbands provisionally designated as tubal epithelial protein 1(TEP-1) and tubal epithelial protein 2 (TEP-2), was observedin explants of the Fallopian tube mucosa obtained from eachanatomical site throughout the ovarian cycle (n = 20). TEP-2was not apparent in tubal mucosa obtained from post-menopausalwomen (n = 5). De-novo synthesis of TEP-1 and TEP-2 was notapparent in autoradiographs of radiolabelled proteins from short-termexplants of endometrium. From the autoradiographs the molecularmass of TEP-1 and TEP-2 was calculated to be 25 kDa and 17 kDa,respectively. Incorporation of glucosamine into newly synthesizedprotein occurred in TEP-2 but not TEP-1. TEP-1 was observedto be immuno-chemically identical to immunoglobulin x lightchains.     

Pregnancy: Function of the corpus luteum, the endometrium and the trophoblast after treatment of tubal pregnancy by prostaglandin F2{alpha}

The activity of the corpus luteum, the endometrium and the trophoblastwas studied after local medical treatment of 31 women with tubalpregnancy. We measured the serum concentration of progesterone,the secretory endometrial protein placental protein 14 (PP14),and human chorionic gonadotrophin (HCG) before and after treatmentby injection of prostaglandin F₂ into the site of the gestationand into the corpus luteum. There was no significant differencein the pre-treatment serum progesterone and serum PP14 concentrationsof 26 women who were treated successfully and of five women,who were operated on after failure of the treatment. After theprostaglandin treatment the serum progesterone and PP14 concentrationsdecreased simultaneously with the serum HCG concentration orremained at a low, constant concentration. We conclude thatmeasurement of serum progesterone and PP14 cannot be used forselection of patients for treatment by prostaglandin F₂ or formonitoring the effect of the treatment. The injection of prostaglandininto the ovary has either no effect on the activity of the corpusluteum or induces only a partial luteolysis.     

The expression of {alpha}V, {alpha}5, {beta}1, and {beta}3 integrin chains on ejaculated human spermatozoa varies with their functional state

Evidence has been presented suggesting the involvement of integrinsand their ligands in mammalian fertilization. In this studywe asked whether the     

Localization and variable expression of G alpha(i2) in human endometrium and Fallopian tubes

BACKGROUND: Heterotrimeric G proteins take part in membrane-mediated cell signalling and have a role in hormonal regulation. This study clarifies the expression and localization of the G protein subunit G alpha(i2) in the human endometrium and Fallopian tube and changes in G alpha(i2) expression in human endometrium during the menstrual cycle. METHODS: The expression of G alpha(i2) was identified by Polymerase chain reaction (PCR), and localization confirmed by immunostaining. Cyclic changes in G alpha(i2) expression during the menstrual cycle were evaluated by quantitative real-time PCR. RESULTS: We found G alpha(i2) to be expressed in human endometrium, Fallopian tube tissue and in primary cultures of Fallopian tube epithelial cells. Our studies revealed enriched localization of G alpha(i2) in Fallopian tube cilia and in endometrial glands. We showed that G alpha(i2) expression in human endometrium changes significantly during the menstrual cycle, with a higher level in the secretory versus proliferative and menstrual phases (P < 0.05). CONCLUSIONS: G alpha(i2) is specifically localized in human Fallopian tube epithelial cells, particularly in the cilia, and is likely to have a cilia-specific role in reproduction. Significantly variable expression of G alpha(i2) during the menstrual cycle suggests G alpha(i2) might be under hormonal regulation in the female reproductive tract in vivo.     

Progressive rise in the expression of interleukin-6 in human endometrium during menstrual cycle is initiated during the implantation window

In order to be prepared for implantation, human endometriumundergoes a predictable series of proliferative and secretorychanges. Cytokines play an important role in regulation of thesechanges. Therefore, in this study, we immunolocalized the cytokine,interleukin-6 (IL-6), its receptor and the signal transducergp130 in human endometrium throughout the menstrual cycle. Duringthe entire menstrual cycle, the IL-6 receptor and gp130 werefound primarily in the endometrial glands and to a lesser extentin the stroma. The immunoreactivity of these proteins did notchange in endometrial cells during the entire menstrual cyclewith an exception of reduced immunoreactivity of gp130 in endometrialglands during menstrual phase. Immunostaining showed that immunoreactiveIL-6 was weakly expressed in human endometrium during the proliferativephase. Strong immunoreactivity for IL-6 appeared in endometriumduring the putative 'implantation window'. Expression was byfar most pronounced both in the glandular and surface epithelialcells. The amount of immunoreactive IL-6 in the epithelium progressivelyincreased during the secretory/menstrual phases. During thelate secretory phase, only stromal cells in the upper functionalisexhibited immunoreactivity for IL-6. Western blot analysis corroboratedthe immunohistochemical data. Human endometrial IL-6 consistedof a protein with an apparent mobility of 26 kDa. The immunoreactiveband of IL-6 was weak in the proliferative phase. The expressionof this protein increased progressively during the secretory/menstrualphases. The findings show a cell-specific pattern of distributionfor immunoreactive IL-6 in human endometrium. The menstrualcycle-dependent expression of IL-6 suggests that this cytokinemay play a role in changes in endometrium that prepare thistissue for implantation and menstrual shedding.     

Immunohistological distribution of the secretory endometrial protein, 'pregnancy-associated endometrial {alpha}2-globulin', a glycosylated {beta}-lactoglobulin homologue, in the human fetus and adult employing monoclonal antibodies

We have previously demonstrated that pregnancy-associated endometrial₂-globulin (₂-PEG), the human glycosylated (-lactoglobulin homologue(HG-BLG), is quantitatively the major secretory soluble proteinproduct of the secretory endometrium during the latter halfof the menstrual cycle and decidua spongiosa of the gestationalendometrium during early pregnancy, and is principally localizedto the glandular epithelium. In the present study employingmonoclonal antibodies in immunohistological techniques, thedistribution and localization has been examined in normal andpathological tissues of the adult and first-trimester fetus.No significant staining for ₂-PEG was detected in any non-reproduction-associatedtissue in the normal adult nor any tissue in the fetus. In theadult, most intense staining was associated with the endometrialglandular epithelium in the uterus or in ectopic sites in patientswith endometriosis. During the menstrual cycle and pregnancy,appearance of ₂-PEG in endometriosis was strongly linked withits appearance in uterine endometrial tissue, suggesting thatendometriotic tissue exhibited competence to respond to thesame hormonal milieu required to induce synthesis in the uterineendometrium. Localization to the mucosal epithelium of the Fallopiantube was consistent with synthesis of ₂-PEG, albeit at low levels,and staining at this site reflected fluctuations of stainingwithin the uterus. Of the pathological specimens examined, stainingwas only detected in a proportion of ovarian carcinomas. Nostaining was detected in the mammary gland, a site of -lactoglobulinsynthesis, whether obtained during pregnancy or lactation. Theseobservations support the proposal that during the menstrualcycle and pregnancy, the endometrial glandular epithelium representsthe major source of the glycosylated -lactoglobulin homologueand serum measurements may be employed to assess its functionalpresence. Mechanisms of regulation of its production that wouldaccount for the histological observations are discussed.     

Clomiphene citrate does not affect the secretion of alpha3, alphaV and beta1 integrin molecules during the implantation window in patients with unexplained infertility.

BACKGROUND: The expression of integrin molecules on the endometrium suggests that certain integrins may participate in the cascade of molecular events leading to successful implantation. A prospective, controlled study was carried out to investigate the effect of clomiphene citrate (CC) on secretions of beta1, alpha3 and alphaV integrin molecules in the endometrium of patients with unexplained infertility during the implantation window. METHODS: A total of 40 endometrial samples was evaluated in both spontaneous (n = 13) and ensuing clomiphene-treated cycles (100 mg on days 5-9) and also from fertile women serving as controls (n = 14) during postovulatory 7th or 8th day of menstrual cycle. A semiquantitative grading system (H-score) was used to compare the immunohistochemical staining intensities. Endometrial thickness and serum oestradiol and progesterone concentrations were also measured on the day of sampling. RESULTS: Staining of alpha(v) but not beta1 and alpha3 integrins was significantly less intense in infertile cases than fertile control cases (1.42 +/- 0.12 versus 2.21 +/- 0.13 respectively, P = 0.012) and this was not restored to normal concentrations with treatment. CONCLUSIONS: Our study indicated that cc treatment significantly decreased the endometrial thickness and increased oestradiol and progesterone concentrations. However, secretion of alpha(v), beta1 and alpha3 integrin molecules, which might play a role in implantation, was not affected.     

The effect of different hormone therapies on integrin expression and pinopode formation in the human endometrium: a controlled study

BACKGROUND: Integrin expression and pinopode formation have been proposed as a means of distinguishing receptive endometrium from non-receptive in clinical practice, thus offering new directions for the development of contraceptive approaches targeted to the endometrium as well as a better understanding of occult causes of infertility in women. The aim of the present study was to investigate the effect of ovulation induction, oral contraception, treatment with dehydrogesterone, and different regimens of hormone replacement therapy on endometrial alphavbeta3 integrin expression and pinopode formation using a prospective, controlled study design. METHODS: Histological dating, alphavbeta3 integrin expression and pinopode formation were evaluated in control and treated cycles in six groups of women including eight subjects per group and who received clomiphene citrate, ovarian stimulation for IVF, oral contraception, dehydrogesterone for endometrial luteal phase defect, or two different regimens of hormone replacement therapy. Twelve healthy fertile women served as a general control group. RESULTS: Alphavbeta3 integrin expression and pinopode formation in the human endometrium were closely related to endometrial maturation as defined by histological dating and this was irrespective of endometria being in-phase or out-of-phase and the hormonal treatment received. Only for clomiphene citrate did a direct effect with reduction in pinopode formation in the midluteal phase seem to exist. CONCLUSION: Alphavbeta3 integrin expression and pinopode formation in the human endometrium are processes closely related to endometrial maturation and this is irrespective of endometria being in-phase or out-of-phase and the hormonal treatment received. The potential usefulness of those two so-called endometrial markers of implantation as targets for contraceptive approaches or fertility-promoting strategies seems unlikely.     

Uterus and endometrium: The three-dimensional organization of the smooth musculature in the ampulla of the human Fallopian tube: a new morpho-functional model

The three-dimensional organization of the smooth musculaturearound the human ampulla is revealed by means of scanning electronmicroscopy after NaOH maceration and ultrasonic microdissectionof the interstitial connective tissue. The muscular wall ofthe ampulla appears as a continuous network of randomly anastomosedsmooth muscle cell bundles that showed a multidirectional arrangement.The smooth muscle cell bundles modify their orientation alongtheir course, intertwine repeatedly with each other and dichotomize,generating new bundles with a different orientation from thatat the origin. These results demonstrate that the myosalpinxof the human ampulla is not organized into clear cut longitudinally,circularly or spirally arranged layers, as suggested in previouslight microscopy studies. In contrast, the presence of a networkof multidirectional smooth muscle cell bundles revealed in thisstudy suggests that there is no morphological evidence for unidirectionalperistalsis, and that the musculature is probably structurallydesigned to stir rather than push the tubal contents. Thesemorphological findings better explain the random pattern ofpropagation of the contraction waves and the electrical impulsesthrough the smooth musculature of the human ampulla, as postulatedin early experimental physiological studies. Further, they suggesta specific function for the ampullar musculature which may notbe only strictly related to tubal content transport.     

Expression of IgA and secretory component in the normal and in adenocarcinomas of Fallopian tube, endometrium and endocervix

The occurrence and localization of IgA and secretory components (SC) were examined in the normal and in adenocarcinomas of Fallopian tube, endometrium and endocervix. IgA-containing immunocytes were identified in the stroma of 90% of normal Fallopian tubes. It is suggested that the Fallopian tube may have an immunological function and may, together with the endocervix, constitute the local secretory immune system of the female genital tract. IgA and SC were frequently demonstrated in the cytoplasm and luminal secretion of adenocarcinomas of the endocervix, endometrium and Fallopian tube. This study has shown a decrease in immunoreactivity of SC among poorly differentiated adenocarcinomas but has failed to demonstrate any correlation between the expression of IgA and the degree of differentiation of the tumours. Secretory component appears, therefore, to be more useful than IgA as an indicator of secretory activity and differentiation of adenocarcinomas of the female genital tract.     

Differential expression of the metalloproteinase MMP3 and the alpha5 integrin subunit in human myometrium at labour

Extensive tissue remodelling occurs in the human myometrium before, during and after parturition. The aim of this study was to investigate the expression of two tissue remodelling molecules, matrix metalloproteinase 3 (MMP3) and alpha5 integrin (ITGA5) subunit in human myometrium, during pregnancy and labour. mRNA and protein were isolated from human pregnant labouring and non-labouring myometrial tissue, and also from human primary uterine smooth muscle cells (SMCs). Semi-quantitative RT-PCR, real-time fluorescence RT-PCR and western blotting were subsequently performed to determine the expression levels of MMP3 and ITGA5 in the myometrial tissues during pregnancy and labour, and in the primary uterine SMCs. The expression of MMP3 and ITGA5 mRNA and protein are reported for the first time during pregnancy and labour in human myometrium. Furthermore, a significant increase in expression of MMP3 mRNA (41-fold, P = 0.001), and a significant decrease in ITGA5 mRNA expression (4-fold, P < 0.001) at labour, were observed. Protein expression of these two molecules was also altered at labour, pro-MMP3 protein expression significantly increased while ITGA5 protein expression decreased, with labour onset. Expression of these molecules was also observed in primary cultured human uterine SMCs. The differential expression of these two tissue remodelling molecules at labour and their detection in uterine SMCs highlights their potential importance in myometrial function during pregnancy, labour and post-partum.