11 research outputs found
Pathogenesis of Ectopic Pregnancy: Is there a Role for the Endocannabinoid System in Modulating Embryo-Tubal Transport?
No full textBackground: The molecular mechanisms of ectopic pregnancy remain unclear. Studies from knockout mice suggest that perturbations in oviductal endocannabinoid levels, endocannabinoid receptors (CB1) or endocannabinoid degrading enzyme (fatty acid amide hydrolase, FAAH) expression result in infertility secondary to physical trapping of embryos in their oviducts. Perturbations in endocannabinoid metabolism and action may therefore underlie ectopic pregnancy.
Aims: To (1) quantify plasma and tubal endocannabinoid levels (2) evaluate blood activity of FAAH and the endocannabinoid degrading enzyme N-acylphosphatidyl-ethanolamine phospholipase-D (NAPE-PLD) and relate that to β-hCG levels (3) evaluate the expression of cannabinoid receptors (CB1, CB2), FAAH and NAPE-PLD in Fallopian tubes and (4) examine the effect of endocannabinoids [N-arachidonoylethanolamine (AEA), N-oleoylethanolamide (OEA) and N-palmitoylethanolamide (PEA)] on cilia beat frequency (CBF) in tubal epithelial cells ex-vivo.
Methods: Whole blood collected from women with ectopic pregnancy and suitable controls were used for quantification of endocannabinoids by UHPLCMS/MS and FAAH and NAPE-PLD activity by HPLC methods. Fallopian tubes were fixed in formalin for immunohistochemistry and had RNA and protein extracted for RT-qPCR and immunoblotting respectively. Fallopian tube explants were exposed to endocannabinoids and changes in CBF evaluated using highspeed digital camera.
Results: Plasma AEA, OEA, PEA and tubal AEA were significantly higher (p < 0.05) in ectopic pregnancy compared to controls. Tubal OEA and PEA showed a similar trend though the results were not statistically significant. Blood FAAH but not NAPE-PLD activity was attenuated (p < 0.05) in ectopic pregnant women consistent with the higher endocannabinoid level observed in plasma. CB1 and FAAH were localised in Fallopian tube and showed significant attenuation (p < 0.05) in ectopic pregnancy compared to luteal phase controls. Exposure of Fallopian tube epithelial cells to OEA unlike methanandamide and PEA resulted in a significant reduction (p < 0.05) in CBF.
Conclusion: The results implicate ECS dysfunction in the pathogenesis of ectopic pregnancy
Laparoscopic surgery for ovarian pregnancy using diathermy hook with conservation of ovary
No full textA 31-year-old woman presented with a 7-week history of irregular vaginal bleeding without abdominal pain. She had been using the intrauterine contraceptive device (IUD) for the last 3 years. A pregnancy test was positive and subsequent serum beta human chorionic gonadotropin (beta-HCG) was 4992 mIU/mL. A transvaginal ultrasound scan demonstrated an empty uterus with an associated adnexal mass but no free fluid. A right primary ovarian ectopic pregnancy was diagnosed a laparoscopy. This was managed laparoscopically using monopolar diathermy hook with conservation of the ovary and minimal blood loss. Ovarian pregnancy is rare, especially in women without the classical risk factors for tubal pregnancy, and efforts should be made to exclude ectopic pregnancy in the absence of abdominal pain in a woman of reproductive age presenting with prolonged and irregular vaginal bleeding. Methods to conserve the ovary are also encouraged in cases of ovarian pregnancy
Variation in stability of endogenous reference genes in fallopian tubes and endometrium from healthy and ectopic pregnant women
No full textRT-qPCR is commonly employed in gene expression studies in ectopic pregnancy. Most use RN18S1, β-actin or GAPDH as internal controls without validation of their suitability as reference genes. A systematic study of the suitability of endogenous reference genes for gene expression studies in ectopic pregnancy is lacking. The aims of this study were therefore to evaluate the stability of 12 reference genes and suggest those that are stable for use as internal control genes in fallopian tubes and endometrium from ectopic pregnancy and healthy non-pregnant controls. Analysis of the results showed that the genes consistently ranked in the top six by geNorm and NormFinder algorithms, were UBC, GAPDH, CYC1 and EIF4A2 (fallopian tubes) and UBC and ATP5B (endometrium). mRNA expression of NAPE-PLD as a test gene of interest varied between the groups depending on which of the 12 reference genes was used as internal controls. This study demonstrates that arbitrary selection of reference genes for normalisation in RT-qPCR studies in ectopic pregnancy without validation, risk producing inaccurate data and should therefore be discouraged
The Role of Ultrasound Simulation in Obstetrics and Gynecology Training: A UK Trainees’ Perspective
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From Fertilisation to Implantation in Mammalian Pregnancy—Modulation of Early Human Reproduction by the Endocannabinoid System
No full textThere is an increasing recognition that the endocannabinoid system is the crucial cytokine-hormone system regulating early human pregnancy. The synchronous development of the fertilized embryo and the endometrium to ensure timely implantation has been shown to be one of the pivotal steps to successful implantation. This development is thought to be regulated by a finely balanced relationship between various components of the endocannabinoid system in the endometrium, the embryo and the Fallopian tube. In addition, this system has also been shown to be involved in the regulation of the development and maturation of the gametes prior to fertilization. In this review, we will examine the evidence from animal and human studies to support the role of the endocannabinoid system in gametogenesis, fertilization, implantation, early pregnancy maintenance, and in immunomodulation of pregnancy. We will discuss the role of the cannabinoid receptors and the enzymes involved in the synthesis and degradation of the key endocannabinoid ligands (e.g., anandamide and 2-arachinoylglycerol) in early reproduction
Selection of suitable endogenous reference genes for relative gene expression studies in human spermatozoa using quantitative real-time PCR
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