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Study Question: Can of Clinical Genetics, Maastricht University Medical Centre, Maastricht kisspeptin and its analogues regulate the motility of human decidual stromal cells and what intracellular signaling pathways are involved?, Summary Answer: Kisspeptin analogue-mediated cell motility in human decidual stromal cells via the focal adhesion kinase (FAK)-steroid receptor coactivator (Src) pathway suggesting that kisspeptin may modulate embryo implantation and decidual programming in human pregnancy., What Is Known Already: The extravillous trophoblast invades the maternal decidua during embryo implantation and placentation. The motile behavior and invasive potential of decidual stromal cells regulate embryo implantation and programming of human pregnancy., Study Design, Size, Duration: Human decidual stromal cells were isolated from healthy women undergoing elective termination of a normal pregnancy at 6- to 12-week gestation, after informed consent., Participants/materials, Setting, Methods: Kisspeptin analogues were synthetic peptides. Cell motility was estimated by an invasion and migration assay. Immunoblot analysis was performed to investigate the expression of kisspeptin receptor and the effects of kisspeptin analogues on the phosphorylation of FAK and Src. Small interfering RNAs (siRNAs) were used to knock down the expression of kisspeptin receptor, FAK, Src, matrix metallo-proteinases (MMPs) 2 and 9, and extracellular signal-regulated protein kinase (ERK) 1/2., Main Results and the Role of Chance: The kisspeptin receptor was expressed in human decidual stromal cells. Kisspeptin agonist decreased, but antagonist increased, cell motility. Kisspeptin agonist decreased the phosphorylation of FAK and Src tyrosine kinases, whereas antagonist increased it. These effects on phosphorylation were abolished by kisspeptin receptor siRNA. The activation of cell motility by kisspeptin analogues was suppressed by siRNA knockdown of endogenous FAK (decreased 66%), Src (decreased 60%), kisspeptin receptor (decreased 26%), MMP-2 (decreased 36%), MMP-9 (decreased 23%), and ERK 1/2 inhibitor (decreased 27%)., Limitations, Reasons for Caution: Human decidual stromal cells were obtained from women having terminations after 6-12 weeks of pregnancy and differences in timing could affect their properties., Wider Implications of the Findings: Kisspeptin acting within the endometrium has a potential modulatory role on embryo implantation and decidual programming of human pregnancy., Study Funding/competing Interest(s): This work was supported by grant NSC-104-2314-B-182A-146-MY2 (to H.-M.W.) from the Ministry of Science and Technology, Taiwan, and grants CMRPG3E0401 and CMRPG3E0402 (to H.-M.W.). This work was also supported by grants from the Canadian Institutes of Health Research to P.C.K.L. P.C.K.L. is the recipient of a Child & Family Research Institute Distinguished Investigator Award. The authors have no conflicts of interest to disclose., Trial Registration Number: N/A., (© The Author(s) 2019. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

The interleukin (IL)-1 system is a major regulator of local cellular interactions during embryonic implantation. Because IL-1beta and IL receptor antagonist (IL-1ra) are both expressed in human endometrium, we hypothesized that an appropriate ratio of IL-1beta to IL-1ra might favor the process of embryo implantation. Therefore, we investigated IL-1 regulation of the quantitative ratio of IL-1beta/IL-1ra messenger RNA (mRNA) expression in human endometrial stromal cells using quantitative competitive PCR, as well as intracellular protein expression after stromal cell solubilization. Confluent stromal cell cultures were stimulated with human IL-1beta (0-1000 IU/mL) for 24 h. After 24 h, total RNA was extracted, reverse transcribed, and coamplified by PCR with a defined amount of internal standard. The quantitative ratio was determined by the density of target to the internal standard. After culture with IL-1beta for 24 and 48 h, stromal cells were solubilized, and the intracellular protein levels of IL-1beta and IL-1ra were measured by enzyme-linked immunosorbent assay. The IL-1beta and IL-1ra mRNA were both up-regulated, and IL-1R tI mRNA was down-regulated, by IL-1beta in a dose-dependent manner. The quantitative ratio of IL-1beta to IL-1ra mRNA was constant with the presence of increasing concentrations of IL-1beta (1-1000 IU/mL). IL-1beta and IL-1ra protein was not detected in conditioned media of cultures before addition of IL-1beta. IL-1beta and IL-1ra protein levels increased with increasing amounts of IL-1beta after solubilization of stromal cells. The IL-1beta was detectable after 12 h of culture, in comparison with IL-1ra, which was detectable after 24 h of IL-1beta stimulation. These results suggest that IL-1 may play a crucial role in embryo-maternal interaction by regulating stromal cell expression of IL-1beta and IL-1ra, resulting in an appropriate ratio during the process of embryonic implantation.

Indirect inguinal hernia is neglected by many female patients. When women undergo laparoscopy for infertility work-up or other gynaecological conditions, the hernia appears pressurized by the pneumoperitoneum. A large opening on the peritoneum followed by the patent canal of Nuck-as seen in the patient in this case report-is accompanied by a bulge above the labia major. This situation was treated with simple ligation of the canal of Nuck from the internal inguinal ring, and the vulvar bulging mass quickly disappeared, even while under pneumoperitoneal pressure. Laparoscopic hernia repair is currently performed with a mesh prosthesis to cover the defect; however, in female patients it seems unnecessary to use such an invasive method designed largely for males. According to our experience, the concomitant simple closure method for female indirect inguinal hernia is a quick and simple procedure, and deals with the problem during the same laparoscopy. In addition, disappearance of the bulging mass under the pneumoperitoneum offers a useful diagnostic test to secure hernia management.

The case report of a young woman with recurrent pelvic desmoid tumour successfully treated with tamoxifen is described. The desmoid tumour recurred within 6 months after the initial exploratory laparotomy. Tamoxifen therapy led to complete relief of ascites within 2 months and complete tumour regression by the end of the fourth month, and the patient has remained stable for 6 years. Without sacrificing pelvic organs or major vessels and preserving reproductive ability, tamoxifen should be considered as the first drug of choice in such a recurrent condition.

The purpose of this study was to investigate the possible regulatory roles of insulin-like growth factors (IGF) and IGF binding protein-1 (IGFBP-1) on postmenopausal endometrium in women undergoing hormone replacement therapy (HRT). Endometrial tissues and blood samples were collected from 25 postmenopausal women with abnormal uterine bleeding before and after HRT. As a control, blood and endometrial samples were also obtained at mid-luteal phase (days 18-23 of the menstrual cycle) from 10 women with benign uterine leiomyoma during surgical intervention. Expression of mRNA for IGF-I, IGF-II, type 1-IGF receptor, IGFBP-1, oestrogen receptor (ER) and progesterone receptor (PR) in the endometrium was explored by a semiquantitative reverse transcription-polymerase chain reaction (RT-PCR). Serum concentrations of IGFBP-1, oestradiol, progesterone, follicle stimulating hormone (FSH) and sex hormone-binding globulin (SHBG) were also determined. In the endometrium obtained from postmenopausal women with abnormal uterine bleeding (before HRT), expression of IGFBP-1 was undetectable, whereas PR expression was abundant. HRT significantly up-regulated expression of IGFBP-1 but down-regulated PR. Moderate expression of IGF-I, IGF-II, type 1-IGF receptor and ER was identified in the postmenopausal endometrium before HRT. HRT significantly down-regulated IGF-I, up-regulated IGF-II, but had no effects on expression of type 1-IGF receptor and ER. In conclusion, up-regulation of IGFBP-1 collaboratively with down-regulation of IGF-I may account for the protective effect of progesterone in HRT on the endometrium.

The aim of this study was to investigate the second trimester concentrations of maternal urine human chorionic gonadotrophin beta-core fragment (HCGbetacf) in Asian pregnanci2es with fetal chromosomal abnormalities. HCGbetacf concentrations were analysed from 34 urine samples in chromosomally abnormal pregnancies, including 28 cases of Down's syndrome, one case of trisomy 18, and five cases of other chromosomal abnormalities (one mosaic deletion and four translocations), and in a cohort of 268 normal pregnancies receiving second trimester amniocentesis. Results were normalized to urine creatinine (Cr) concentration and converted to the multiple of the median (MOM) concentration for the appropriate gestation. The median HCGbetacf MOM concentrations of Down's syndrome pregnancies (12.89) was significantly higher than that of normal pregnancies (1. 06) (P < 0.00001). Wide variations of HCGbetacf concentrations were observed in other chromosomally abnormal pregnancies. There were 18 of 28 (64%) Down's syndrome cases but one of five (20%) other chromosomally abnormal cases with HCGbetacf concentrations above the 95th centile of the control values (8.22 MOM cut-off). These findings suggest that HCGbetacf could be a potential marker in urine screening for fetal Down's syndrome in Asians.

Maldevelopment of the Müllerian duct system may result in various urogenital anomalies including didelphic uterus with a hypoplastic cervix and obstructed hemivagina. We report a patient with this anomaly who was treated by laparoscopic hemi-hysterectomy and hysteroscopic resection of hemivagina. A 17 year old patient who had complained of vaginal pus-like discharge on and off for 1 year was diagnosed by MRI to have a double uterus with obstructed right hemivagina and ipsilateral renal agenesis. After hysteroscopic identification of hypoplasia of the right uterine cervix, laparoscopic resection of the right uterus and right Fallopian tube and hysteroscopically assisted resection of the vaginal septa were performed successfully. From our experience, combined laparoscopy and hysteroscopy may be an efficacious alternative in the management and diagnosis of Müllerian anomalies.

A case of laparoscopic management of an ectopic pregnancy in a previous Caesarean section scar is reported. A 30 year old woman was admitted to our hospital for profuse vaginal bleeding 2 weeks after an abortion had been performed. A urine pregnancy test was positive. Abdominal ultrasound revealed a well-encapsulated bulging mass over the lower anterior uterine wall measuring 7x5 cm. Hysteroscopy revealed retained gestational tissue in the lower corpus despite a normal uterine cavity. An incision was made over the most prominent area of the mass by operative laparoscopy. Dark reddish tissue suggestive of the products of conception was removed using grasping forceps. One-layer of continuous endoscopic sutures along the affected uterine wall was made with 1-0 Prolene. Laparoscopy enabled the successful treatment of an unruptured ectopic pregnancy in a previous Caesarean scar and made it possible to preserve the patient's reproductive capability.

In patients with agenesis of the vagina and cervix but with a functional endometrium, the traditional treatment is hysterectomy with construction of a neovagina. We report successful treatment by laparoscopically assisted full thickness skin graft for reconstruction in a patient with congenital agenesis of the vagina and uterine cervix concomitant with haematometra and ovarian endometrioma in a 12 year old girl. Postoperatively, the vaginal skin graft healed well, and menstruation first appeared 4 weeks later. In our opinion, a combined laparoscopic and vaginal procedure with full thickness skin graft is an efficacious alternative in managing such genital defects.

To determine the risks when the primary methotrexate (MTX) treatment of cervical pregnancy has an unsatisfactory outcome, we conducted a Medline search on relevant literature published from January 1983 to June 1997. The search yielded 28 publications of 48 cases of cervical pregnancy. These and four new cases from our institutions were used in our study. A cervical pregnancy that presented with a serum beta-human chorionic gonadotrophin concentration of > or = 10,000 mIU/ml [odds ratio (OR) 10.82, 95% confidence interval (CI) 2.59, 45.14], gestational age at > or = 9 weeks (OR 6.44, 95% CI 1.46, 28.52), embryonic cardiac activity (OR 14.29, 95% CI 2.95, 76.92), and crown-rump length of >10 mm (OR 13.33, 95% CI 1.46, 120.48) was considered to be associated with a higher unsatisfactory rate of primary MTX treatment. A concomitant feticide was found to enhance the therapeutic effect of MTX treatment if embryonic cardiac activity was evident (OR 0.13, 95% CI 0.02, 0.68). Administration of a high dose of MTX did not seem to be more effective than a lower one. Our findings supported some previous observations and, more importantly, provided useful clinical information in selecting appropriate candidates for MTX treatment in cases of cervical pregnancy.

Interleukin-1 (IL-1) is expressed in human endometrium and has been shown to play an integral role in local cellular interactions during implantation. In addition, the matrix metalloproteinase (MMP) and its inhibitor, the tissue inhibitor of metalloproteinase (TIMP), are crucial during implantation, mediating in vitro trophoblast penetration, and are regulated by several cytokines expressed by trophoblast cells. We have investigated the roles of IL-1 beta and transforming growth factor-beta (TGF beta) in regulating TIMP-1, TIMP-3, and 92-kDa type IV collagenase messenger ribonucleic acid (mRNA) expression in human endometrial stromal cells using quantitative competitive PCR. Confluent stromal cell cultures treated with progesterone and estradiol for 9 days were stimulated with IL-1 beta, IL-1 beta plus anti-IL-1 beta antibody, TGF beta, and TGF beta plus anti-TGF beta antibody for an additional 24 h. Competitive complementary DNA fragments were constructed by deletion of a defined fragment from each of the target complementary DNA sequences and coamplified in quantitative competitive PCR as an internal standard. TIMP-1 and TIMP-3, but not 92-kDa type IV collagenase mRNA, were expressed in stromal cells. The 92-kDa type IV collagenase mRNA was only expressed after stimulation with IL-1 beta. IL-1 beta both augmented 92-kDa type IV collagenase mRNA expression and decreased TIMP-1 and TIMP-3 mRNA expression in a dose-dependent manner. Conversely, TGF beta augmented TIMP-1 and TIMP-3 mRNA expression, but did not affect 92-kDa type IV collagenase expression. IL-1 and TGF beta-mediated changes were both neutralized by specific antibodies. These results provide indirect evidence that IL-1 and TGF beta may play crucial roles at the embryo-maternal interface during trophoblast invasion by regulating stromal cell expression of TIMP-1, TIMP-3, and 92-kDa type IV collagenase, all of which are known to be important in trophoblast invasion.

Chylous ascites is an extremely rare complication of laparoscopic presacral neurectomy (LPSN), and treatment is still controversial. Four patients undergoing LPSN for dysmenorrhoea or chronic pelvic pain were complicated with chylous ascites. Two were successfully treated with bipolar cauterization and one, after the failure of initial treatment by bipolar cauterization, was then effectively managed by compression with Gelform and closure of the peritoneum of the presacral area by suture through laparoscopy. The fourth patient had persistent chyle leakage from the drainage tube after electrocauterization and was finally cured by conservative management including removal of the drainage tube and a low-fat diet for 3 weeks. Chylous ascites has not been reported in laparoscopic presacral neurectomy. Management that is quick, effective and subjects the patients to the least amount of suffering is still unresolved. Repeated laparoscopy can be considered to identify the possibility of injury to lymphatic vessels, to relieve abdominal distention due to chyle accumulation, and to apply electrocauterization or compression with Gelform and closure of the peritoneum. Conservative treatment with a low-fat diet may need a longer time. The use of a drainage tube may provide negative pressure allowing a continuous leakage of chyle. However, more controlled study is required to identify the most proper and effective management.

A 36 year-old infertile female developed a stage IV (FIGO) ovarian carcinoma consisting of a poorly differentiated Sertoli-Leydig cell tumour after receiving one course of ovulation induction with follicle stimulating hormone (FSH), human menopausal gonadotrophin (HMG) and human chorionic gonadotrophin (HCG) followed by gonadotrophin-releasing hormone analogue (GnRHa). The patient died of liver metastasis and hepatic failure 4 1/2 months after first diagnosis, despite aggressive treatment consisting of debulking surgery and aggressive adjuvant chemotherapy.

Seven patients with secondary amenorrhoea were diagnosed as having severe uterine synechiae by hysterosalpingography (HSG) and hysteroscopy, which revealed short, narrow and/or scarred uterine cavities as cone or column shapes. Laminaria tents were used to distend the uterine cavity prior to transcervical resectoscopy to completely dissect the dense adhesions. A more traditional postoperative management included an intrauterine device, oestrogen and antibiotics. Transcervical resectoscopy combined with laminaria appears to be a safe and effective means of restoring the uterine cavity. All seven patients not only achieved normal menstruation but also normal uterine cavity as confirmed by subsequent HSG and hysteroscopy. In addition, three patients thereafter became pregnant, two of whom have had successful term deliveries.

The case reports of 22 patients with cervical pregnancies treated by methotrexate (MTX) administration and published in English literature between 1983 and 1995 were reviewed, by either original paper review or follow-up under international collaboration, to determine the subsequent reproductive performance and obstetric outcomes. Out of 22 cases, 18 (78%) MTX chemotherapy attempts succeeded with complete remission and four (22%) failed. Of the 13 women who wished to conceive and could be followed for at least 3 years, nine succeeded in having live births without congenital malformations, one spontaneously aborted and three suffered infertility. In general, MTX chemotherapy alone or combined with adjuvant methods such as subsequent cervical curettage or cervical tamponade, or intracervical potassium chloride injection, appears to be a convenient and effective method for the treatment of the majority of cervical pregnancies before 12 weeks gestation, and has not been shown to have detrimental effects on subsequent reproductive capacities, obstetric outcomes and progeny health for those cases with successful preservation of the uteri.

Despite the expanding role of laparoscopic surgery in many gynaecological fields, some discrepancies still exist regarding the efficacy of laparoscopic myomectomy in treating patients with large symptomatic leiomyoma. In this report, a better operative procedure and the results of treatment are evaluated. Patients (n = 14) presenting with infertility, menorrhagia, pressure symptoms or pelvic mass associated with a large leiomyoma were managed with laparoscopic myomectomy using airlift gasless laparoscopy. Uterine size ranged from 14 to 24 weeks gestational age and the weight of the myoma ranged from 246 to 669 g (mean 454); operative time ranged from 78 to 165 min (mean 104) and blood loss from 90 to 580 ml (mean 201). No major complication occurred during the operation or follow-up. All except one patient were discharged within 72 h of the operation and resumed normal activity within 1 week. When myomectomy is indicated, the airlift gasless laparoscopic approach appears to offer a better alternative to abdominal or pneumoperitoneum laparoscopic surgery in selected cases. Airlift gasless laparoscopy has several advantages: (i) small abdominal incisions and minimal endoscopic equipment are required; (ii) the excised leiomyomata mass can be easily cut into strips and removed through the small abdominal incision; (iii) the uterine defect can be more efficiently repaired using easily performed suture techniques; (iv) high-pressure irrigation and large-volume suction devices can be used without fear of decompressing the pneumoperitoneum; and (v) the potential risk of metabolic and haemodynamic derangements during pneumoperitoneum laparoscopy are obviated. Gasless laparoscopy also has some disadvantages. The exposure obtained with gasless laparoscopy is not as good, under some circumstances, as that achieved by pneumoperitoneum. For patients who are thin, and even those with moderate obesity, the exposure obtained with airlift mechanical suspension is adequate; however, morbidly obese patients with previous abdominal surgery with suspected pelvic adhesions can incur some problems during the operation because of a poor operative field.

Co-culturing embryos on helper cells can mimic the in-vivo environment, thereby enhancing embryo development in vitro. Insulin-like growth factors (IGF) and their binding proteins (IGFBP) also enhance embryo development. To investigate the kinds of IGFBP produced by various cell monolayers and the effects of IGFBP-3 on mouse embryo co-culture systems, 2-cell ICR mouse embryos were cultured in either human tubal fluid medium alone or in the presence of Vero cells, human oviductal cells or endometrial cells. The helper cells were analysed immunohistochemically to investigate the types of IGFBP produced by various cell monolayers. The concentrations of IGF-I and IGFBP-3 in media obtained from the culture of embryos alone, cells alone or cells plus embryos were determined by radioimmunoassays. On day 7, more blastocysts hatched in the co-culture groups (73% in the Vero cell group, 76% in the endometrial cell group and 74% in the oviductal cell group) than in the control group (43%) (P < 0.0001). The results of immunohistochemistry revealed that (i) all three cell groups produced a lot of IGFBP-1, -2 and -3, but only a little of IGFBP-4 and -5; and (ii) IGFBP-1, -2, and -3 were present in blastocysts in either the presence or absence of helper cells. The IGF-I secreted by cell monolayers or embryos was undetectable (detection limit 0.83 microg/l). The IGFBP-3 concentrations in media obtained from co-cultured embryos and cells were significantly higher than in media without embryos (median values in oviductal cell culture medium, 165 versus 127 microg/l, P = 0.04; median values in endometrial cell culture medium, 277.5 versus 183.5 microg/1, P = 0. 0002; median values in Vero cell culture medium, 219 versus 120 microg/l, P = 0.011). Although IGFBP-3 concentration in the medium that contained embryos alone was undetectable by radioimmunoassay (detection limit 1.1 microg/l), immunohistochemistry demonstrated the presence of IGFBP-3 in the embryos. Co-culture in systems in which there was an increased production of IGFBP-3 led to an improved development of mouse embryos. IGFBP can improve the binding of IGF to cell surface receptors of target tissue, and thus enhance the effect of limited IGF concentrations in promoting embryo development in a co-culture system. We conclude that Vero cells, human endometrial cells and oviductal cells produce IGFBP-1, -2, -3, -4 and -5. IGFBP-3 may play a role in embryotrophic potential by either regulating the action of IGF or directly enhancing embryo development.

Construction of a neovagina using pelvic peritoneum via a laparoscopically assisted approach was used in 18 patients with congenital absence of the vagina. A better operative procedure is reported, which was modified from our preliminary technique, and the results of treatment are evaluated. Pelvic peritoneum was used for construction of a vagina, replacing a laparotomy by a minimally invasive laparoscopic approach. During follow-up, the advantages of our procedures are: (i) minimal likelihood of 'poor take' or later contraction because an autograft peritoneal epithelial line is used; (ii) minimal short and long term care is required; (iii) the technique is simple in experienced hands and has all the well-recognized benefits of minimal invasive surgery; (iv) the average length of neovagina is adequate and patency and depth can be maintained with minimal dilatation; (v) the neovagina, with epithelial lining resembling that of a normal vagina, facilitates comfortable sexual intercourse; (vi) the procedure is unaccompanied by dyspareunia or scarring, which are frequently associated with other techniques; (vii) less emotional stress and better cosmetic and economic benefits are noted.

We report on a case of ureteral injury during laparoscopically radical hysterectomy to treat a patient with a stage IA2 carcinoma of the uterine cervix. The advantage of laparoscopically radical hysterectomy is the prevention of the identification and dissecting of the ureter from a vaginal approach, the main problem in the Schauta technique. However, ureteral injury may still occur while resecting the cardinal ligament without good visualization of the ureter during a vaginal procedure. Because the ureter was well identified and freed laparoscopically, a vaginal approach after uterine removal may be an option for treatment. Moreover, using the ureteral stent as a marker during a modified Schauta technique is helpful in the prevention of ureteral injury during laparoscopically assisted radical vaginal hysterectomy.

Laparoscopic rescue after tubal anastomosis failure is reported for the first time. The patient was a 33 year old woman sterilized by Pomeroy's method. Reconstruction of fertility was achieved by mini-laparotomy with isthmo-isthmic end-to-end anastomosis of bilateral tubes. Unfortunately, the patient did not become pregnant and tubal occlusion was diagnosed 6 months later by hysterosalpingography. Laparoscopic rescue with repeat isthmic-ampullary anastomosis and adhesiolysis was performed. Post-operatively, the patient had one menstruation and then achieved an intrauterine pregnancy. Instead of in-vitro fertilization/embryo transfer, laparoscopic rescue may be an alternative option for the patient with failed anastomosis.

From July 1992 to May 1994, 31 cases of obstructive azoospermia-induced infertility underwent 35 epididymal sperm aspiration procedures and assisted reproductive techniques. These included in-vitro fertilization (IVF), zygote intra-Fallopian transfer (ZIFT), subzonal insemination (SUZI) and intracytoplasmic sperm injection (ICSI) with embryo transfer or tubal embryos transfer. The motile spermatozoa were retrieved using a gauge 24 Medicut cannula and flush medium. Total oocyte fertilization rate was 23.4% and the clinical pregnancy rate was 12.5% (four out of 32 treatment cycles). Congenital absence of the vas deferens was found in 16 cases (51.6%) and secondary genital duct obstruction was present in 15 cases (48.4%). In all, 29 aspirations were retrieved from the caput of the epididymis and six from the epididymal body or tail. The fertilization rate for the caput spermatozoa was much less than that for other areas of the epididymis (P < 0.05). Though there were no predictable differences in fertilization rates between the congenital and acquired groups, by using epididymal spermatozoa for assisted reproduction, the congenital group seemed to have a stronger tendency to achieve pregnancy (20 versus 5.9%).

Ovarian preservation before abdominal irradiation may be recommended for young patients with various types of invasive cancer. The most common site for ovarian transposition is just below the iliac crest or posterolateral to the uterus. Here, we demonstrate laparoscopic ovariopexy with an automatic stapling device to transpose the ovaries to an uncommon site, i.e. lower anterolateral abdomen, in a patient with medulloblastoma prior to her receiving irradiation of the craniospinal axis. The anterolateral transposition of the ovary not only kept it away from the irradiated field but allowed it also to be shielded by the uterus. Thus, ovarian function could be well preserved.

Leuprolide acetate (Lupron) belongs to the group of gonadotrophin-releasing hormone agonists (GnRHa). It is used in conjunction with human menopausal gonadotrophin (HMG) and/or follicle stimulating hormone (FSH) for ovarian stimulation in the so-called ultra-short, short or long protocols. In our programme, the long protocol is mainly used, which starts routinely with s.c. administration of Lupron, 1 mg/day, from day 21 of the cycle preceding ovum retrieval. After pituitary desensitization is achieved, gonadotrophin administration is started. However, unexpected, spontaneous pregnancy may occur during the flare-up phase of Lupron administration. We present six pregnancies and four deliveries which have occurred under these circumstances. The mechanisms by which pregnancy may occur and escape from luteolytic, abortifacient and teratogenic effects of prolonged use of GnRHa are discussed, as well as pregnancy outcome.

Our objective was to evaluate and compare the efficacy and mechanisms of co-culturing mouse embryos with Vero cells in both Dulbecco's modified Eagle's medium/Ham's F12 (DMEM/F12) and human tubal fluid (HTF) culture medium. Two-cell CB6F1 mouse embryos were cultured either in the presence of Vero cells (group A) or in culture medium alone (group B). In DMEM/F12 significantly more morulae developed in group A than in group B on day 3 (91 versus 23%; P less than 0.01). In contrast, the mouse embryos grew rapidly in HTF and significant differences were noted only in later embryonic stages (on day 5; 86% and 50%, P less than 0.01 of group A and B respectively, hatching or hatched). Similar experiments using DF1 and ICR mouse strains also revealed enhanced embryo development in the presence of Vero cells. To determine whether the embryo-enhancing effects of Vero cells were due to the removal of toxins or to the secretion of embryotrophic factors, ICR mouse embryos were cultured in fresh media with cells (group A), without cells (group B) and in cell-free conditions using cell-conditioned media which were obtained in the presence (group C) or absence (group D) of embryos. These results demonstrated that completion of hatching was highest (52%; P less than 0.01) in group A after 6 days in culture. There were no significant differences between groups B, C and D (rates of total hatching 18, 17 and 17%, respectively). It is concluded that Vero cells improve the development of mouse embryos and this is likely to be due to removal of substances inhibitory to development.(ABSTRACT TRUNCATED AT 250 WORDS)

The lymphocyte pore-forming protein (PFP)--perforin, also named cytolysin--is a potent mediator of cytotoxicity found in the granules of cytotoxic T lymphocytes and natural killer (NK) cells. Granulated metrial gland (GMG) cells found in the pregnant mouse uterus express perforin and are thought to be highly activated cytolytic lymphocytes related to NK cells. Their role in pregnancy is unknown. Human endometrial granulocytes (EGs) are phenotypically similar to murine GMG cells and, like them, express NK cell markers. However, up to now it was not known whether EGs also express perforin. By means of immunohistochemical analysis, using antisera specific for perforin and monoclonal antibodies to CD56 (NKH-1), CD2 (T11), CD3 (Leu-4), CD4 (Leu-3a), and CD8 (OKT-8), we demonstrated that perforin is present in EGs in the decidualized endometrial stroma and decidual tissue of first-trimester gestational endometrium. In fact, double immunohistochemical labeling demonstrated the co-expression of perforin and NKH-1. This population also expressed some T-cell surface antigens (Leu-4 and T11), but not Leu-3a or OKT-8. Chorionic villi, in contrast, lack perforin+ cells. The presence of a potent cytolytic mediator in NK-like cells in both murine and human pregnant uterus raises the issue of the function of such cells in pregnancy.

The polymerase chain reaction (PCR) was used to identify human papillomavirus (HPV) in cervicovaginal cells in normal individuals and in patients with cervical intraepithelial neoplasia (CIN). By use of a set of primers encoding the E6 region of the HPV genome, the presence of HPV DNA was demonstrated in the cervicovaginal cells of 43 (42.2%) of 102 normal individuals and in all 12 CIN patients. High sensitivity of the PCR method produced an additional 9 positive results on second sampling from 48 individuals who were initially HPV-negative. On the other hand, 26 (24.3%) of 107 HPV-positive individuals were HPV-negative when sampled a second time 5-7 days later. The data suggested that retest probably should be considered for patients clinically suspected of having HPV infection whose initial test results are negative for HPV DNA. Also, single HPV DNA-positive results should be accepted with caution.