Indoleamine-dioxygenase is expressed in human decidua at the time maternal tolerance is established

The semi-allogeneic fetus has to be tolerated by the maternal immune system. In mice, it has been shown that inhibiting indoleamine-dioxygenase (IDO) leads to fetal rejection, suggesting a central significance for IDO in establishing maternal tolerance. Consequently, we have analyzed IDO expression in human endometrium and decidua to determine whether it may be of significance in human reproduction. Endometrial (n=60) and decidual (n=68; first and second trimester) tissue samples and isolated cells were analyzed for IDO mRNA and protein expression by real-time PCR, Western blot and immunohistochemistry. IDO expression in the decidua of proven fertile women (n=34) was compared to women presenting with their first pregnancy (n=22) and women with a history of miscarriages (n=12). Expression of IDO was localized in glandular epithelial cells and scattered stromal leukocytes. Expression started at the mid-luteal phase in the menstrual cycle and was high until the second trimester of pregnancy. However, glandular expression of IDO decreased during the second trimester, whereas expression in villous trophoblast started at this time. There were no significant differences in decidual IDO expression between proven fertile women and women presenting with their first pregnancy or women with a history of miscarriages. From the expression pattern we conclude that IDO may play a central role in human pregnancies for the establishment of maternal tolerance of fetal antigens. Thereby, IDO expression may be needed in each pregnancy independently from prior pregnancies, and a history of miscarriage may not reflect a general deficiency in IDO expression.     

Induction of infertility in adult male bonnet monkeys by immunization with phage-expressed peptides of the extracellular domain of FSH receptor

Active immunization of proven fertile adult male bonnet monkeys (Macaca radiata) with phage-expressed follicle-stimulating hormone receptor (FSHR)-specific peptides from the extracellular domain resulted in a progressive drop in sperm count with all animals becoming azoospermic by day 100. However, serum testosterone concentrations were unaltered during the entire course of study and animals exhibited normal mating behaviour. Breeding studies with proven fertile female monkeys revealed that all the immunized males were infertile. Following interruption of immunization on day 225, sperm counts returned to normal with restoration of fertility. These results indicate that infertility can be induced in adult male monkeys by interfering with the action of FSH using specific peptides of the extracellular domain of FSHR as antigens, without the risk of producing cross-reacting antibodies to the other glycoprotein hormones.     

Deoxyribonucleic acid synthesis in mesometrial arteries of guinea pigs during oestrous cycle, pregnancy and treatment with oestradiol benzoate

Terminal portions of mesometrial arteries which supply the myometrium and the placenta, respectively, were excised at various stages of the ovarian cycle and pregnancy, as well as after oestrogen treatment, and incubated in solutions containing [3H]thymidine. The incorporation of radioactivity into the acid-insoluble material was measured in order to evaluate the actual rate of DNA synthesis before and during pregnancy and after oestrogen administration. We found that the rate of incorporation rises 30-fold before oestrus and 40-fold in the beginning of pregnancy. A 70-fold rise in incorporation was initiated by 10 micrograms oestradiol benzoate (administered subcutaneously) in dioestrous and ovariectomized animals. We conclude that similar peaks of DNA synthesis in mesometrial arteries occur during oestrus and at the beginning of pregnancy. Oestradiol is capable of inducing the observed surge of DNA synthesis. The results are in agreement with the concept that oestradiol is a chemical regulator of arterial growth during pregnancy.     

Effect of a specific estrogen antibody on pregnancy establishment in the bonnet monkey (Macaca radiata)

The requirement for estrogen for pregnancy establishment has not been conclusively demonstrated in primates. Selective neutralization of estrogens was achieved in mated female monkeys during preimplantation and postimplantation periods by injecting characterized estrogen antiserum from either day 14 to 18 or day 28 to 32 of cycle. While estrogen deprivation during preimplantation period in 5 animals exposed to 14 ovulatory cycles resulted in only one pregnancy, only 3 of 13 monkeys treated during postimplantation period continued pregnancy to term. In comparison with controls (4 of 5 monkeys becoming pregnant), the percent protection against pregnancy in animals treated during preimplantation period was 93. The pregnancy termination in 10 of 13 monkeys treated during postimplantation period when compared with normal postimplantation pregnancy wastage in our colony (2%) is also highly significant (P less than 0.01). The present study demonstrates a critical need for estrogen during the peri-implantation period for a successful pregnancy establishment in primates.     

Total urinary oestrogen excretion during normal menstrual cycle and normal pregnancy in Nigerians.

Total oestrogens were measured during normal menstrual cycles and during pregnancy in healthy Nigerian women. During the menstrual cycle, maximum excretion was consistently observed in the luteal phase in contrast to findings in European and American populations. During pregnancy, oestrogen excretion was similar to that reported amongst Europeans only in early pregnancy; oestrogen excretion did not show the usual rapid rise in late pregnancy.     

Regulation of expression and localisation of the Na+/H+ exchanger (NHE) 3 and the NHE regulatory factor 2 in baboon placental syncytiotrophoblast by oestrogen

Our understanding of the regulation of the expression of the sodium hydrogen exchangers (NHE) and their regulatory factors (NHERF), which play important roles in fetal-placental homeostasis, is incomplete. We previously showed that the expression and localisation of NHE3 and NHERF2 in the juxtanuclear compartment of the placental syncytiotrophoblast were markedly decreased between mid and late baboon pregnancy. In the current study, immunocytochemical fluorescence localisation and level of NHE3/NHE1 and NHERF1/NHERF2 proteins were determined in late gestation in baboons untreated or treated throughout the second half of gestation with an aromatase inhibitor CGS 20267 alone (reduced oestrogen levels by >95%) or with oestradiol to determine whether oestrogen regulated antiporter developmental expression. The immunocytochemical expression of NHE3 and NHERF2 in the juxtanuclear compartment was minimal in baboons untreated or treated with CGS 20267 plus oestradiol (i.e. oestrogen-replete) but extensive in oestrogen-suppressed animals. Moreover, the abundant expression of NHERF2 in fetal vascular endothelium of oestrogen-replete baboons was decreased in oestrogen-suppressed animals. In contrast, expression and localisation of NHE1 and NHERF1 in the placental syncytiotrophoblast were not altered by oestrogen deprivation in baboons. Based on our current and previous findings, we propose that oestrogen plays an important role in regulating localisation and expression of components of the NHE system within and consequently development and function of the primate placental syncytiotrophoblast.     

Cryopreservation of human spermatozoa: correlations of ultrastructural sperm head configuration with sperm motility and ability to penetrate zona-free hamster ova

Freezing and storage of human male gametes is associated with a reduction in the overall semen quality and establishment of pregnancy. This study was done to evaluate the integrity of sperm head ultrastructure (SHU) with computerized and vapor freezing. Comparisons were made between the effect of cryopreservatives glycerol (G) and dimethylsulfoxide (DMSO) on SHU. Twelve ejaculates from five proven fertile donors were studied with the use of routine semen analysis, zona-free hamster ova, and SHU. Both cooling processes, regardless of the preservative used, significantly reduced sperm function and the number of SH with intact plasma membranes. The staged cooling technique was substantially superior to vapor freezing in all parameters analyzed (P less than 0.01). G was less detrimental to the postthaw SHU than 1 M DMSO. A significant positive correlation (r = 0.98; P less than 0.01) was noted between the total number of intact SH and motile sperm. Computerized freezing in a G-diluted semen rendered a sperm environment that allowed the highest number of forms with intact SH membranes and with the best chances to penetrate zona-free hamster ova.     

A Review of 50 Cases of Primary Amenorrhoea

Summary: Fifty cases of primary amenorrhoea are reviewed with reference to the aetiology. In all patients the buccal smear chromatin complement was measured and chromosomal study proceeded to if this revealed less than 25% of cells to be chromatin positive. Total urinary gonadotrophin and oestrogen assays were performed in all patients and the spectrum of values seen is presented.
The most common cause was chromosomal abnormality (24%). Although investigation to the extent performed is often not necessary to establish a precise diagnosis, such study is indicated to assess gonadal potential and subsequent fertility potential. In the present series, 31 (62%) patients were potentially fertile. With rational and physiological therapy all patients should be capable of leading normal, sexually active lives. To the present time, pregnancy has resulted from gonadotrophin therapy in the 5 patients in whom it has been indicated.     

Oestrous cycle and pregnancy effects on the distribution of oestrogen and progesterone receptors in bovine endometrium

Reproductive cyclicity and pregnancy require cell and tissue specific expression of oestrogen (OR) and progesterone receptors (PR). To determine the effects of cycle day and early pregnancy (days 16-30) on the distribution of OR and PR, uteri were collected from cycling (n=39), pregnant (n=14), and steroid-treated ovariectomized cattle (n=10). Immunohistochemistry was used to localize OR and PR, and staining intensity was scored. Moderate staining for the PR was detected in the stroma at proestrus (days 17-20). Expression increased through oestrous, reaching maximal levels by metestrus (days 1-6) (P< 0.05). During diestrus (days 7-16), PR expression was downregulated (P< 0.05). Progesterone receptor expression was detected in the luminal epithelium in one of three animals each at days 3 and 6 of the oestrous cycle. In pregnant animals only, caruncular stroma showed reduced PR staining compared to intercaruncular stroma (P< 0.05). Reactivity to the OR antibody in cycling animals was detected in the stroma and glandular epithelium. Levels increased through proestrus and oestrous, were maximal during metestrus (P< 0.05), and decreased during diestrus (P< 0.05). Interestingly, there was transient upregulation in the luminal epithelium on day 16 or day 14 (P< 0.05). This was not evident at day 16 of pregnancy, although some OR reactivity was present in stroma and glands. By day 18 of pregnancy OR were undetectable (P< 0.05). Oestrogen-treated ovariectomized animals had increased OR and PR expression in comparison to progesterone treated cows (P< 0.05). Untreated ovariectomized cows showed constitutive steroid receptor expression. These results support the widely accepted role for sex steroids in the regulation of their receptors and indicate that there is paracrine regulation of steroid receptor expression in the bovine uterus.     

Sperm DNA fragmentation is a novel biomarker for early pregnancy loss

Research questionSpontaneous pregnancy loss affects 10–15% of couples, with 1–2% suffering recurrent pregnancy loss and 50% of miscarriages remaining unexplained. Male genomic integrity is essential for healthy offspring, meaning sperm DNA quality may be important in maintaining a pregnancy. Does sperm DNA fragmentation measured by alkaline Comet assay act as a biomarker for early pregnancy loss?DesignSperm DNA fragmentation was measured by alkaline Comet test in 76 fertile donors and 217 men whose partners had recently experienced miscarriage. Couples were divided into five groups for analysis: one miscarriage after spontaneous conception; two or more miscarriages after spontaneous conception; one miscarriage after fertility treatment; two or more miscarriages after fertility treatment and biochemical pregnancy.ResultsReceiver operator characteristic curve analysis was used to determine ability of the average Comet score (ACS), low Comet score (LCS) and high Comet score (HCS) to diagnose miscarriage and develop clinical thresholds comparing men whose partners have miscarried with men with recently proven fertility. Male partners of women who had miscarried had higher sperm DNA damage (ACS 33.32 ± 0.57%) than fertile men (ACS 14.87 ± 0.66%; P < 0.001). Average Comet score, HCS and LCS all have promise as being highly predictive of sporadic and recurrent miscarriage using clinical thresholds from comparisons with fertile men's spermatozoa: receiver operating characteristic curve AUC for ACS ≥26%, 0.965; LCS ≤70%, 0.969; HCS ≥2%, 0.883; P <0.0001.ConclusionsSperm DNA damage measured by the alkaline Comet has promise as a robust biomarker for sporadic and recurrent miscarriage after spontaneous or assisted conception, and may provide novel diagnoses and guidance for future fertility pathways.     

Folate-metabolizing gene variants and pregnancy outcome of IVF

There is growing evidence that folate status and variation in folate-metabolizing genes are involved in female reproductive functions. This study evaluated the influence of maternal blood folate, vitamin B(12), homocysteine and 10 folate pathway gene variants on IVF outcome. Also, the prevalence of these polymorphisms was compared in 439 female IVF patients and 225 fertile controls. MTHFR 677 CT heterozygotes had a higher proportion of good-quality embryos and an increased chance of pregnancy. MTHFR 1793 GA heterozygosity was associated with a lower percentage of previously failed IVF treatments. Heterozygosity for FOLR1 1816 C/delC and 1841 G/A was associated with a raised risk of pregnancy loss. The CTH 1208 GT genotype was associated with an increased chance of pregnancy and a smaller number of previously failed IVF cycles and the genotype frequency was lower in IVF patients with three or more previously failed IVF treatments compared with fertile controls. SLC19A1 80 GA heterozygotes had a decreased number of previously failed IVF treatments and were more prevalent among fertile controls. In conclusion, polymorphisms in folate-metabolizing genes may affect ovarian stimulation and pregnancy outcome of IVF, and heterozygous individuals, rather than the wild-type homozygotes, appeared to have more favourable outcomes.     

妊娠期宫颈癌筛查

妊娠期宫颈癌是严重影响母儿健康的围妊娠期合并症。了解妊娠期的生理特点与宫颈癌的关系,掌握不同妊娠期宫颈癌的特点及筛查方法,建立临床医师对妊娠期宫颈癌的筛查规范,加强妊娠期宫颈癌的早期筛查,对减少育龄妇女及围产儿病死率具有重要意义。     

Estimation of fertility and fecundity in women receiving artificial insemination by donor semen and in normal fertile women

Summary. The pregnancy rates after artificial insemination by donor semen (AID) have been compared with pregnancy rates in normal fertile women to assess the efficiency of AID. To do this, the curve y = a (1−(1−b) ^x ) was fitted to life-tabled cumulative pregnancy rates. The equation describes a model in which the parameter a is the proportion of women who are potentially fertile under the conditions of treatment, and in which the parameter b is the pregnancy rate per cycle (or fecundity) of these fertile women. For 259 AID patients with no previous pregnancy a was 65% while for 57 AID patients with a previous pregnancy after AID 'a' was 99.9%. The values of b were similar for the two groups of patients, being 20% and 22% respectively. Women without fertility problems who had become pregnant after discontinuing oral contraception provided the reference group. Since only pregnant women were selected, a was 100% by definition. The values of b for the reference group were 22% for 100 primigravid women and 20% for 100 multigravid women. Only 65% of the AID patients were potentially fertile with AID, but those that were fertile became pregnant at the same rate as normal women who discontinued oral contraception.     

Comparative Study of Total Urinary Oestrogen and First Morning Urinary Oestrogen in Monitoring Gonadotrophin Therapy

Total 24-hour urinary oestrogen has been used extensively and proven to be reliable in monitoring gonadotrophin therapy for induction of ovulation. However, the method is time consuming for the patient and incomplete collection, as expected, is not uncommon, hence interfering with the treatment result. Oestrone-3-glucuronide in first morning urine samples has been shown to correlate well with plasma oestradiol-17 beta levels during normal menstrual cycles. A comparative study was made to examine the correlation between the levels of 24-hour urinary oestrogen and first morning urinary oestrogen in patients on gonadotrophin therapy. Creatinine levels were also determined to exclude the factor of variable excretion of oestrogen. The correlation thus found is highly significant (p less than 0.001). Hence first morning urinary oestrogen to creatinine ratios may be used to replace 24-hour urinary oestrogen in biochemical monitoring of gonadotrophin therapy.     

Pregnancy rates in sequential in vitro fertilization cycles by oocyte donors(1)

OBJECTIVE: To evaluate the clinical outcome of in vitro fertilization (IVF) treatment cycles from individual oocyte donors who underwent multiple sequential donations. METHODS: We reviewed clinical outcome data from sequential anonymous oocyte donation cycles using donors who underwent multiple IVF stimulations. Donors were grouped by the interval between cycles and the cycle number (rank). The primary outcome measure was delivery rate by individual donor per retrieval from the combined derivative fresh and frozen embryo transfers. RESULTS: Duration and amount of gonadotropin therapy and the fertilization rates did not correlate significantly with the interval between cycles or cycle rank. Cumulative delivered pregnancy rates for cycles 1-6 were 51.5%, 54.6%, 50.5%, 51.5%, 51.1%, and 57.6%, respectively. Delivered pregnancy rates did not vary by interval between cycles. CONCLUSION: Young healthy presumed or proven fertile women can reliably donate oocytes for at least six cycles with the expectation of consistently high pregnancy rates.     

不明原因重复性自然流产妇女配偶精子DNA完整性

目的:探讨精子DNA完整性与重复性自然流产(RSA)的关系。方法:85例不明原因RSA妇女配偶(RSA组)和50例已生育的成年健康男性(对照组)的精液,应用精子染色质扩散实验(SCD)检测精子DNA完整性。将RSA组根据1年后怀孕结果分为3个亚组:怀孕组(30例)、流产组(26例)、未孕组(29例)。结果:RSA妇女配偶DNA损伤精子的百分率(14.6±6.9)%与对照组的(12.9±3.8)%相比无统计学差异(P0.05)。DNA损伤精子百分率大于20%视为精子DNA完整性异常,则有17.6%的RSA患者配偶的精子DNA完整性异常,6%的正常生育男性精子DNA完整性异常,但差异无统计学意义(P0.05)。怀孕组、流产组、未孕组配偶的DNA损伤精子百分率分别为(12.4±5.3)%,(14.6±6.5)%和(16.8±8.1)%,未孕组与对照组比较,差异有统计学意义(P0.05),怀孕组、流产组与对照组比较,差异无统计学意义(P0.05)。结论:精子DNA完整性异常与RSA继发不育有关,有必要筛查RSA患者配偶的精子DNA完整性。     

Surgical adhesives and tubal sterilization: an experimental study

AIM: Our aim was to assess the application of three currently used surgical adhesives in the tubal lumen of rabbits, to promote sterilization, using a transvaginal approach. METHODS: Fifty-seven female albino New Zealand rabbits (114 uterine tubes), which became pregnant and delivered before the experiment, were divided into four groups: GS (sham-24 tubes), GEFIBRI (0.25 mL of fibrin adhesive in 30 tubes), GE-GRF (0.25 mL of resorcin adhesive in 30 tubes) and GEBUTYL (0.25 mL of n-butyl-2-cyanoacrylate adhesive in 30 tubes). The animals were mated with proven fertile males after the experiment and observed over 30, 90 and 180 days. Pregnancy and patency were macroscopically evaluated. The tubal diameter, tubal mucosa, myosalpinx, total optical density and inflammatory process were microscopically evaluated. The statistical analysis was performed by McNemar and Wilcoxon tests for the subgroups, and Fisher's exact test and Kruskal-Wallis test for the groups, the differences identified by Dunn's multiple comparisons test (P=5%). RESULTS: GS showed patency and pregnancies in all subgroups. GEFIBRI showed patency and pregnancies in all subgroups. GE-GRF did not show patency or pregnancies, but was associated with severe inflammatory process and tubal morphology alterations. GEBUTYL did not show patency, pregnancies or morphological tubal mucosa alterations. CONCLUSIONS: The n-butyl-2-cyanoacrylate adhesive effectively promoted tubal obstruction, did not cause tubal morphological alterations, nor did it impair the rabbit pregnancy. The fibrin adhesive failed to cause the occlusion. The GRF adhesive, in spite of producing tubal occlusion, caused severe uterine tubes damage.     

Adipositas und Schwangerschaft

Obesity is a major worldwide health problem. The rate of obese women of fertile age is rising continuously. An end to this trend is not in sight. In fact, the direct and indirect associations of obesity and adverse pregnancy outcomes such as gestational diabetes, hypertension in pregnancy, thromboembolism, preterm birth and an increased rate of fetal malformations have been proven in hundreds of studies. Moreover birth itself may be significantly more frequently complicated by severe complications such as shoulder dystocia, prolonged labour, tears and lacerations and is associated with higher rates of caesarean sections. Nevertheless extreme obesity represents an extreme socio-economic burden by an estimated fivefold increase of pregnancy-associated health care costs. Long-term effects such as early onset juvenile diabetes in the offspring of obese mothers seem to be generally underestimated so far. Obstetricians and gynaecologists appear to be one key to address preventive measures (i.e. diet counselling, health programmes) in obese mothers and their families.     

Repeat ectopic pregnancy: a study of 123 consecutive cases.

In a study of 1,330 ectopic pregnancies, 123 (9 per cent) patients experienced repeat tubal pregnancies. Of these 123 patients with repeat ectopics, 96 (78 per cent) were surgically sterilized with their second operative procedure. The remaining 27 potentially fertile patients have had nine pregnancies, but only three living children. Cornual wedge resection of the uterine tube predisposed in interstitial pregnancy and to rupture of the uterus in four cases. Four patients had three ectopic pregnancies each. Repair of the involved tube is usually technically possible following linear salpingotomy and should be done if future fertility is desired. If the patient does not desire future pregnancy, she should be sterilized to avoid the risk of repeat ectopic pregnancy.     

育龄女性垂体泌乳素微腺瘤手术前后内分泌改变及疗效分析

目的:探讨育龄女性中垂体泌乳素微腺瘤患者微创手术治疗的疗效。方法:回顾性分析显微手术治疗的育龄女性垂体泌乳素微腺瘤24例,对手术前后内分泌改变及疗效进行分析。结果:本组24例全部手术全切除肿瘤,术后复查血清PRL,下降至正常水平23例。术后当月月经恢复正常6例;术后6月月经恢复正常18例。其中1例术后d3恢复月经,且术后3个月经B超查证实怀孕。13例溢乳患者均于治疗后2-16周内停止溢乳。结论:显微手术切除可作为育龄女性中症状性垂体泌乳素微腺瘤的首选治疗方法。