Category Archives: Human Reproduction

BACKGROUND

Thin-section oblique axial magnetic resonance imaging (MRI) is useful in staging endometrial and cervical carcinomas but there are no data on its contribution to assessing deep endometriosis. We evaluated the contribution of this MRI technique to diagnosis of uterosacral ligament (USL) endometriosis.

METHODS

In this retrospective study, two radiologists, who were blinded to the surgical and histological results, compared the results from conventional sagittal and axial MRI with those from conventional plus thin-section (3 mm) oblique axial MRI in 100 symptomatic patients. Descriptive statistical analyses including sensitivity, specificity, positive and negative predictive values, accuracy and positive and negative likelihood ratios were performed. Kappa for inter-observer agreement was calculated.

RESULTS

Conventional MR images for the diagnosis of left/right USL endometriosis revealed accuracies of 69/76 and 59/75%, sensitivities of 66/71 and 52/71% and specificities of 76/86 and 76/82% for senior and junior readers, respectively. The combination of conventional and thin-section oblique axial MR images revealed accuracies of 82/87 and 74/81%, sensitivities of 89/93 and 73/81% and specificities of 61/72 and 76/79%, for senior and junior readers, respectively. When conventional MRI combined with oblique axial T2-weighted MRI was compared with conventional MRI use only, significant differences in diagnostic accuracies were observed for right (P= 0.04) and left (P= 0.01) USL endometriosis.

CONCLUSIONS

Thin-section oblique axial T2-weighted imaging can improve the success of conventional MRI for assessment of USL endometriosis. Further prospective studies are required before this new MR protocol is performed routinely for suspected pelvic endometriosis.

BACKGROUND

Heavy menstrual bleeding and dysmenorrhea are two top complaints from women with symptomatic adenomyosis, yet their etiology is poorly understood. Tissue factor (TF) has been shown to be upregulated in endometriosis and at the endometrial bleeding sites of women with long-term progestin only contraception. We sought to investigate the expression and localization of TF in eutopic and ectopic endometrium of women with adenomyosis and in endometrium of women without adenomyosis. We also sought to determine the relationship, if any, between TF immunoreactivity and the amount of menses, uterus size and severity of dysmenorrhea.

METHODS

We retrieved tissue samples of eutopic and ectopic endometrium from 50 women with adenomyosis and of endometrium from 18 women without adenomyosis. The tissue sections were subjected to immunostaining and microscopic evaluation to assess the presence and localization of TF in both proliferative and secretory phases in both eutopic and ectopic endometrium and normal endometrium. Information on the amount of menses, severity of dysmenorrhea and other information were collected.

RESULTS

We found that TF immunoreactivity was significantly increased in both eutopic and ectopic endometrium as compared with normal endometrium. In addition, we found that the elevated TF immunoreactivity is associated with heavy menses and increased severity of dysmenorrhea.

CONCLUSIONS

These results suggest that TF is involved in adenomyosis-associated heavy menstrual bleeding and dysmenorrhea and thus may be a potential therapeutic target in treating symptomatic adenomyosis and perhaps also chronic pelvic pain in women with adenomyosis.

In 2005, the European Society for Human Reproduction and Embryology (ESHRE) PGD Consortium published a set of Guidelines for Best Practice PGD to give information, support and guidance to potential, existing and fledgling PGD programmes. Subsequent years have seen the introduction of new technologies as well as the evolution of current techniques. Additionally, in light of recent advice from ESHRE on how practice guidelines should be written/formulated, the Consortium believed it was timely to update the PGD guidelines. Rather than one document that covers all of PGD, the new guidelines are separated into four documents, including one relating to organization of the PGD centre and three relating to the methods used: DNA amplification, fluorescence in situ hybridization and biopsy/embryology. Here, we have updated the sections on organization of the PGD centre. One area that has continued to expand is Transport PGD, in which patients are treated at one IVF centre, whereas their gametes/embryos are tested elsewhere, at an independent PGD centre. Transport PGD/preimplantation genetic screening (PGS) has a unique set of challenges with respect to the nature of the sample and the rapid turn-around time required. PGS is currently controversial. Opinions of laboratory specialists and clinicians interested in PGD and PGS have been taken into account here. Current evidence suggests that PGS at cleavage stages is ineffective, but whether PGS at the blastocyst stage or on polar bodies might show improved delivery rates is still unclear. Thus, in this revision, PGS has been included. This document should assist everyone interested in PGD/PGS in developing the best laboratory and clinical practice possible.

Conservative surgical treatment for symptomatic endometriosis is frequently associated with only partial relief of pelvic pain or its recurrence. Therefore, medical therapy constitutes an important alternative or complement to surgery. However, no available compound is cytoreductive, and suppression instead of elimination of implants is the only realistic objective of pharmacological intervention. Because this implies prolonged periods of treatments, only medications with a favourable safety/tolerability/efficacy/cost profile should be chosen. In the past few years, innumerable new drugs for endometriosis, which would interfere with several hypothesized pathogenic mechanisms, have been studied and their use foreseen. However, robust evidence of in vivo safety and efficacy is lacking and, at the moment, the principal modality to interfere with endometriosis metabolism is still hormonal manipulation. Regrettably, in spite of consistent demonstration of a major effect on pain even in patients with deeply infiltrating lesions, progestins are underestimated and dismissed in favour of more scientifically fashionable and up-to-the-minute alternatives. Moreover, oral contraceptives (OCs) dramatically reduce the rate of post-operative endometrioma recurrence and should now be considered an essential part of long-term therapeutic strategies in order to limit further damage to future fertility. Finally, women who have used OC for prolonged periods will be protected from an increased risk of endometriosis-associated ovarian cancer. To avoid the several subtle modalities for distorting facts and orientating opinions in favour of specific compounds, progestins and monophasic OC used continuously are here proposed as the reference comparator in all future randomized controlled trials on medical treatment for endometriosis.

In 2005, the European Society for Human Reproduction and Embryology (ESHRE) PGD Consortium published a set of Guidelines for Best Practice PGD to give information, support and guidance to potential, existing and fledgling PGD programmes. The subsequent years have seen the introduction of new technologies as well as evolution of current techniques. Additionally, in light of recent advice from ESHRE on how practice guidelines should be written and formulated, the Consortium believed it was timely to revise and update the PGD guidelines. Rather than one document that covers all of PGD, the new guidelines are separated into four new documents that apply to different aspects of a PGD programme, i.e. organization of a PGD centre, fluorescence in situ hybridization (FISH)-based testing, amplification-based testing and polar body and embryo biopsy for PGD/preimplantation genetic screening (PGS). Here, we have updated the sections that pertain to FISH-based PGD. PGS has become a highly controversial technique. Opinions of laboratory specialists and clinicians interested in PGD and PGS have been taken into account here. Whereas some believe that PGS does not have a place in clinical medicine, others disagree; therefore, PGS has been included. This document should assist everyone interested in PGD/PGS in developing the best laboratory and clinical practice possible. Topics covered in this guideline include inclusion/exclusion criteria for FISH-based PGD testing, referrals and genetic counselling, preclinical validation of tests, FISH-based testing methods, spreading of cells for analysis, set-up of local IVF centre and transport PGD centres, quality control/ quality assurance and diagnostic confirmation of untransferred embryos.

BACKGROUND

Cryopreservation of ovarian tissue for fertility preservation is based on the ovarian cortex that contains the vast majority of the follicular reserve, while the remaining tissue, the medulla is discarded. The present study describes the development of a gentle method for isolating pre-antral follicles from human ovarian medulla and evaluating its follicular content.

METHODS

Medulla was collected from 40 girls/women aged 3–35 years undergoing cryopreservation of the ovarian cortex. Follicle density was assessed for all patients and pre-antral follicles were isolated from 22 patients. On the basis of the neutral red (NR) staining of follicles and enzymatic digestion with a mixture of Collagenase IV and Liberase Thermolysin Medium, viable pre-antral follicles were isolated.

RESULTS

NR accumulated in follicles resulting in a distinct red staining within the medulla. Follicle density of the medulla varied from 0 to 9824 follicles/gram of medulla and was significantly higher (P< 0.001) in the 3–9-year age group when compared with older groups (10–35 years). Enzymatic digestion combined with follicle identification by NR yielded a high output of isolated and viable pre-antral follicles from medulla, of which, 3607 follicles were collected and classified. The percentage of primordial and growing follicles decreased and increased, respectively, with age (P< 0.0001 and <0.0007).

CONCLUSIONS

Discarded medulla contained a considerable pool of pre-antral follicles, especially in young girls. Our new method allowed the isolation of viable pre-antral follicles from human ovarian medulla and provides a unique opportunity for basic scientific studies and for culture and grafting purposes.