The baboon model (Papio hamadryas) of fetal loss: maternal weight,age, reproductive history and pregnancy outcome

Background Several risk factors are associated with the incidence of human stillbirths. The prevention of stillbirths in women is a pressing clinical problem. Methods We reviewed 402 pathology records of fetal loss occurring in a large baboon (Papio spp.) colony during a 15‐year period. Clinical histories of 565 female baboons with one or more fetal losses during a 20‐year period were analyzed for weight, age, and reproductive history. Results Fetal loss was most common at term (35.57%) and preterm (28.61%) and less common in the first half of gestation (11.20%) and post‐term (5.22%). Greater maternal weight, older age, history of stillbirth and higher parity were independent predictors for stillbirth. An exponential increase in the incidence of fetal loss was observed beginning at age 14 years in baboons. Conclusions Fetal loss and maternal risk factors associated with stillbirths in baboons were similar to those documented in women.     

Recombinant human TNFRSF1A (r-hTBP1) inhibits the development of endometriosis in baboons: a prospective, randomized, placebo- and drug-controlled study

Endometriosis is associated with chronic inflammation, including an increased macrophage activity with increased secretion of cytokines, such as tumor necrosis factor (TNF) or TNF superfamily member 2, previously known as TNFalpha. In the present study, we tested the hypothesis that recombinant human TNFRSF1A (r-hTBP1) can inhibit the development of endometriotic lesions in the baboon, an established model for the study of endometriosis. Endometriosis was induced using intrapelvic injection of menstrual endometrium in 20 baboons with a normal pelvis. In the first part of the study, 14 baboons were randomly assigned to subcutaneous treatment with r-hTBP1, placebo, or GnRH antagonist (positive control). In the second part of the study, menstrual endometrium from 6 baboons was randomly incubated with either PBS or r-hTBP1 before intrapelvic seeding. Video laparoscopy was performed 25 days later to document the number, surface area, and estimated volume of endometriotic lesions and adhesions; to calculate the revised American Fertility Society (rAFS) score and stage; and to confirm the histological presence of endometriosis. In the first part, baboons treated with r-hTBP1 or with Antide (Bachem) had a lower endometriosis rAFS score, a lower surface area and estimated volume of peritoneal endometriotic lesions, and a lower histological confirmation rate compared with controls. Because of less adnexal and cul-de-sac adhesions, the number of baboons with endometriosis of stage II, III, or IV was lower among baboons treated with r-hTBP1 or Antide than among controls. In the second part, the surface area of endometriotic lesions was lower, and less severe endometriosis was observed in r-hTBP1-treated baboons. No hypoestrogenic effects were observed in baboons treated with r-hTBP1. In conclusion, r-hTBP1 can effectively inhibit the development of endometriosis without hypoestrogenic effects in baboons.     

Synthesis and release of polypeptides by the baboon (Papio anubis) uterine endometrium in culture

This study was designed to identify proteins released in culture by the baboon uterine endometrium. Endometrial tissues from cyclic baboons were minced and cultured in the presence of L-[3H]leucine or L-[35S]methionine for 24 h. The culture media and solubilized tissues were analyzed by one- and two-dimensional gel electrophoresis for secretory products that were uterine-specific. The fluorographs of the one- and two-dimensional gels demonstrated that the proteins released into culture media could be divided into two groups. Group I proteins were present throughout the menstrual cycle and showed minor cyclic variations in intensity, and Group II proteins were those that appeared to be hormonally modulated. Group I was comprised of several high molecular weight proteins (Mr greater than 200,000) and at least five additional proteins ranging in molecular weight from 80,000 to 37,000, with isoelectric points (pIs) of 5.1 to 6.0. Group II consisted of a protein (Mr 33,000; pI 7.6) that was observed only during the follicular stages of the cycle and two other groups of proteins (Mr 130,000 and 88,000) that were present during the luteal stage. Western blots of tissue culture media incubated with antibodies against human placental proteins (PP) and prolactin demonstrated that PP4 and PP7 were secreted throughout the cycle while PP12, PP16, and prolactin were only present during the luteal stage of the cycle. Thus, it appears that the baboon uterine endometrium, like that of the human, secretes a wide array of proteins in culture. Our results also suggest that a few of these proteins are immunologically similar. Endometrial differentiation during the menstrual cycle altered the secretion of some proteins, whereas the synthesis of others appeared to be dependent on either estrogen or progesterone and were stage-specific.     

Cytohistologic correlation between AGUS and biopsy-detected lesions in postmenopausal women

OBJECTIVE: To evaluate histologic findings in patients aged 50 and older whose cervical smears revealed atypical glandular cells of undetermined significance (AGUS). STUDY DESIGN: Computerized records spanning a four-year period were retrospectively analyzed. Thirty patients over age 50 had cervical smears interpreted as AGUS and had follow-up biopsies within 12 months following the abnormal smear. The most important histologic diagnosis from the biopsy specimens was correlated with the subcategory of the cervical smear. RESULTS: Five smears interpreted as AGUS, favor reactive, revealed abnormal histology in four cases: three endometrial polyps and one squamous carcinoma. Two smears interpreted as AGUS, favor dysplasia, revealed squamous intraepithelial lesions on biopsy in both cases. Seventeen smears interpreted as AGUS, favor endometrial cells, revealed abnormal histology in 13 cases: 1 endocervical polyp, 6 endometrial polyps, 3 endometrial hyperplasias and 3 adenomyosis. Six patients with smears interpreted as AGUS, unclassifiable, revealed abnormal histology in five cases: two endocervical polyps, one endometrial polyp, one endometrial carcinoma and one ovarian carcinoma. CONCLUSION: The presence of AGUS in cervical smears from women over 50 was highly predictive of abnormal lesions detected by histologic examination. Although three cancers were detected on histologic follow-up, the most common lesions detected were endometrial polyps.     

Patterns of inhibin and FSH localization in endometrium of baboon (Papio anubis) during menstrual cycle and early pregnancy.

Immunoreactive 10.5 KDa moiety of inhibin and hFSH was present in the baboon endometrium during menstrual cycle, early pregnancy and in castrated animals treated with steroid hormones, estrogen and/or progesterone. Endometrial differences during the menstrual cycle altered the intensity of immunostaining of inhibin and FSH. Maximum staining was observed in late luteal phase for both the hormones. In early pregnancy (35th day), the conceptus increased the staining for inhibin in the adjoining endometrial glands. Treatment of castrated animals with steroids for 14 days caused increased staining for inhibin. Maximum staining was observed when treated with estradiol or progesterone, whereas combination of estrogen and progesterone treatment decreased the staining reaction. In conclusion, both inhibin and FSH were localized in baboon endometrium and were under the influence of estrogen and progesterone.     

The estrogen early response gene FOS is altered in a baboon model of endometriosis

Endometriosis, the presence of a functional endometrium outside of the uterine cavity, is associated with infertility. In our simulated model of pregnancy in baboons with experimental endometriosis, hCG infusion fails to induce expression of the immunoregulatory protein glycodelin. To test the hypothesis that the development of endometriosis is associated with an aberrant endometrial immunological environment, we examined the expression of a series of immunoregulatory genes in endometrium from baboons with and without endometriosis. Six months following intraperitoneal inoculation with menstrual endometrium, eutopic endometrium was surgically collected between Days 9 and 11 postovulation. Control endometrium was similarly collected from disease-free animals. Total RNA was extracted, and biotinylated cDNA probes were hybridized to the SuperArray GEArray Q series Th1/Th2/Th3 cDNA array, representing 96 genes. Gene expression levels were determined using ScanAlyze and GEArray Analyzer software. Seven genes were upregulated, including JUND, FOS, CCL11, NFKB1 and others, in the endometrium from baboons with endometriosis compared with the endometrium from disease-free animals; one gene, IL1R1, was downregulated. Quantitative RT-PCR confirmed upregulation of FOS and CCL11 in endometriotic eutopic endometrium. Immunohistochemical analysis revealed altered levels and distribution of FOS protein in the eutopic endometrium of baboons with induced endometriosis. These data suggest that in an induced model of endometriosis an aberrant eutopic immunological environment results in a decreased apoptotic potential and in rapid alterations in endometrial gene expression. We propose that the reduced fecundity associated with endometriosis has a multifold etiology in spontaneous and induced disease.     

Transcervical endometrial biopsy in the baboon: Effects on the menstrual cycle and relations to protein and dry matter content and histology

The baboon has been used increasingly for reproductive studies. While hormonal regulation of the menstrual cycle and ovulation as well as the endocrinology of gestation have been reported, little information is available describing endometrial parameters. It is the purpose of this paper to describe the ease with which repeated transcervical biopsies can be performed, to describe baseline endometrial protein and dry weight data and to demonstrate that the biopsy procedure itself does not significantly affect the baboons' ability to continue normal menstrual cycle function. Endometrial biopsy samples were taken throughout the menstrual cycle under light ketamine anesthesia. Protein and dry weight contents were determined. Endometrial biopsies Protein and dry weight contents were determined. Endometrial biopsies averaged 25 mg (wet weight) and contained 7.54% protein and 16.3% dry matter. The formulas (Y = a + bx) which expressed the linear relationships between wet weight (mg), protein (μg) and dry matter (μg) content and the correlation coefficients (r) were as follows: between wet weight and protein content – wet weight = 5.58 + 10.0 (protein), Sxy = 4.83, r = 0.883; between wet weight and dry weight – wet weight = 1.99 + 7.94 (dry weight), Sxy = 4.52, r = 0.904; between protein and dry weight – protein = 0.446 + 0.446 (dry weight), Sxy = 4.82, r = 0.870. All three linear regression coefficients were statistically significant (P < 0.001). No significant cyclical patterns in either protein or dry matter content were demonstrable throughout the menstrual cycles. The average length of all nonbiopsy cycles was 32.4 ± 2.7 days and 32.8 ± 3.6 days for those in which biopsies were taken. Similarly, follicular and luteal phase lengths for nonbiopsy and biopsy cycles were 15.4 ± 2.3 and 15.5 ± 2.8 days and 16.9 ± 2.2 and 17.2 ± 3.2 days, respectively. The time required for sex-skin swelling to decrease from maximum to minimum during the luteal phase was shorter, but the quiescent stage was equally lengthened. It was concluded that the endometrium of the baboon was easily accessible for study without causing serious alterations in menstrual cycle function. These studies further demonstrate the potential of the baboon as a model o reproductive studies. In fact, the baboon may well the only practical primate model available for endometrial studies.     

Record review of baboons with histologically confirmed endometriosis in a large established colony

Spontaneous endometriosis was diagnosed in 43 baboons over a 14-year period. Thirty-seven have died; five remain alive; one was sold and lost to follow-up. The average age at diagnosis was 17.2 years; 29 (67%) were between 12 and 21 years of age. Fifteen (35%) were diagnosed by biopsy and received surgical excision of the endometriotic tissue; four of these were identified during caesarian section, confirming one prior report of endometriosis in pregnant animals. Twenty-eight (65%) were diagnosed at or shortly preceding necropsy. When diagnosed by a palpable abdominal mass, there was a significantly greater likelihood the animal died or was killed as a result of complications of endometriosis. When diagnosis was at necropsy, there was a significantly greater likelihood that the animal died from causes unrelated to endometriosis. Early identification with surgical removal appears to provide a benefit for both survival and delivering offspring after diagnosis. In twenty-one baboons (49%), endometriosis affected multiple sites within the peritoneal cavity. In the remaining baboons, lesions were more localized. Ovarian involvement was seen in sixteen (37%) of these baboons. This paper is the first to describe significant ovarian involvement in baboons, previously considered a limitation of the usefulness of this species as an animal model. We also describe the first reported endometriosis seeding of an abdominal surgery scar in a baboon. Many of these baboons were middle aged, had few or no offspring, or had evidence of a long duration of uninterrupted menstrual cycles, consistent with risk factors for women. Endometriosis was an incidental finding in 17 (40%) of these baboons, consistent with previous reports of minimal endometriosis as a common asymptomatic finding in baboons and in women. Overall, endometriosis in baboons presents a spontaneously occurring animal model that shares important features with the disease in women and the rhesus macaque.     

The presence of endometrial cells in cervical smears in relation to the day of the menstrual cycle and the method of contraception

The presence of endometrial cells in cervical smears was studied in a large series of women participating in a population screening program for cervical cancer, in relation to different time periods of the menstrual cycle and to the method of contraception practiced. In the total group of women studied, endometrial cells were present in an average of 12% of the cervical smears. In women who were menstruating cyclically, the percentage of cervical smears containing endometrial cells was not age dependent. Only in women over 52 years was a lower number of endometrium-positive cervical smears found: in postmenopausal women, 0.6% of smears were found to contain endometrial cells. In menstruating women, the frequency of endometrial cells in cervical smears was highest during the menses. After day four, through the proliferative phase, the percentages of cervical smears containing endometrial cells markedly decreased. During the secretory phase, an average of 2% of the smears contained endometrial cells; in the premenstrual phase (after day 25), the percentages of endometrial cell-positive smears rose again. When related to the method of contraception practiced, significant differences in the percentages of cervical smears with endometrial cells appeared. In women using oral hormonal contraceptives, the average numbers of smears containing endometrial cells for the whole cycle as well as for each period of the cycle were significantly lower. This phenomenon might be due to endometrial atrophy on the basis of prolonged use of oral hormonal contraceptives. In women wearing an intrauterine device, at any moment the frequencies of smears with endometrial cells present were significantly higher than the values found in women using any other method of contraception or not using contraceptives. The evaluation of cells originating from the endometrium requires considerable experience. The identification of endometrial cells can be made with greater confidence when the cytologist is aware of the exact date of the menstrual cycle and of the impact on the presence of endometrial cells in cervical smears caused by different methods of contraception.     

子宫内膜息肉研究进展

子宫内膜息肉(endometrial polyps,EMP)由子宫内膜腺体和含有厚壁血管的纤维化内膜间质构成,是局部子宫内膜过度增生形成的有蒂或无蒂的赘生物。子宫内膜息肉是最常见的子宫内膜病变之一,临床表现为子宫不规则出血,或月经量增多、不孕、绝经后出血等,也可无明显临床症状。子宫内膜息肉多数属良性病变,但其可恶变性已经被证实。子宫内膜息肉的发病机制目前尚不明确,传统观点认为其与慢性子宫内膜炎症有关,属慢性炎症范畴,即为生物致炎因子及长期反复机械性刺激所致的反应性增生物;近年来随着分子生物学研究的深入,发现子宫内膜息肉的发生与可能与激素调控下增殖与凋亡失衡相关。近来随着宫腔镜检查技术的推广及激素补充治疗人数的增多,子宫内膜息肉的发病率及检出率逐渐增加,加之经刮宫或电切治疗后复发率非常高,因此,子宫内膜息肉越来越受到临床医生的重视,现将子宫内膜息肉的研究进展作一综述。     

子宫腺肌病发病与干细胞关系的研究进展

子宫腺肌病(adenomyosis,AM)是一种常见的严重影响女性身心健康及女性生育并伴有恶性侵袭性的妇科良性疾病。以子宫内膜的腺体和间质异位至深部子宫肌层,并伴随相邻肌细胞增生和肥大为特征,其临床特点特殊,近期统计研究发现其发病率正逐渐升高,而且趋向年轻化。现有研究报道子宫腺肌发病涉及多方面的因素,但具体发病机制尚不明确。近期研究发现子宫内膜存在的基底层干细胞可能与子宫内膜随月经周期发生脱落、增殖相关,其功能紊乱可能导致子宫内膜增生异常性疾病包括子宫腺肌病的发生,子宫内膜干细胞侵入子宫肌层,在局部环境的诱导下增殖、分化或可形成子宫内膜的异位病灶。子宫内膜干细胞的研究可能为子宫腺肌病发病和治疗带来新的思路和希望。本文参考国内外文献,就近年来子宫腺肌病发病机制及与干细胞相关研究进展进行综述并提出展望。     

Risk factors for dystocia in pigtailed macaques (Macaca nemestrina)

Dystocia (difficult labor) is an important component of the management of nonhuman primates and results in significant fetal and maternal morbidity and increased use of veterinary resources. Dystocias can arise from abnormalities of the maternal pelvis or fetus or uncoordinated uterine activity. Although risk factors for stillbirths have been established in nonhuman primates, risk factors for dystocias have not. The objective of this study was to determine maternal and fetal risk factors for dystocia in macaques. Retrospective data were collected from 83 pigtailed macaques (Macaca nemestrina) diagnosed with dystocia. The diagnosis of dystocia was made based on clinical or pathologic evidence. Maternal records of age, reproductive history, experimental history, clinical records, and fetal birth weight and any applicable fetal necropsy reports were reviewed. The gestational age of the fetus, the infant's birth weight, total previous births by the dam, and the proportions of both viable delivery (inverse effect) and surgical pregnancy interventions (direct effect) in the dam's history generated a model that maximized the experimental variance for predicting dystocia in the current pregnancy and explained 24% of the dystocia deliveries. The number of total previous births and proportion of previous cesarean sections accounted for the greatest effect. This model can identify individual dams within a colony that are at risk for dystocias and allow for changes in breeding colony management, more intense monitoring of dams at risk, or allocation of additional resources.     

Eicosanoids production in endometriosis

In order to investigate the production of eicosanoids in human endometrium, myometrium, leiomyoma, adenomyosis, normal ovary, non-endometrial cyst and endometrial cyst, slices of each tissue were incubated. 6-Keto-prostaglandin (PG) F1 alpha, thromboxane (TX) B2, PGF2 alpha and PGE2 concentrations in the incubation medium were measured by direct RIA. 6-Keto-PGF1 alpha production of adenomyosis was significantly higher than that of endometrium, myometrium and leiomyoma, especially in the menstrual phase. The production of eicosanoids in endometrial cyst was significantly higher than that of non-endometrial cyst and normal ovary. These results suggest that endometriosis is associated with increased eicosanoid production in vivo.     

O-9Uncommon subtypes of borderline nuclear changes: cytological features and predictive value

Introduction:  To establish the significance of cytological features which could predict clinically significant endometrial pathology, and therefore guide reporting practice in cervical samples.
Methods:  A retrospective review of SurePath liquid-based cytology (LBC) cervical samples between 2002 and 2006, obtained at screening and colposcopy. These smears contained normal endometrial cells present at inappropriate times of the menstrual cycle, endometrial cells with atypia (borderline change) and with features suspicious / diagnostic of endometrial carcinoma (glandular neoplasia). False negative and false positive cases detected on subsequent histology were also included. The control group comprised negative samples and a few abnormal smears. All smears were randomly assigned and blinded to menopausal status, age, use of oral contraceptive pill and hormone replacement therapy and presence of intrauterine device. Each smear was reviewed for 16 cytologic criteria and a cytological diagnosis was given for each.
Results:  A total of 219 smears were available for review; 137 were negative, out of which 85 contained normal endometrial cells, 41 contained endometrial cells with atypia, 10 contained endometrial cells with features suggestive of adenocarcinoma and 31 contained endometrial cells with features diagnostic of adenocarcinoma. The feature most associated with benign endometrial cells is top hat with central cell condensation. In contrast, the features associated with malignant endometrial cells are smooth nuclear membrane, pale chromatin, small nucleoli and scalloped borders.
Discussion:  The criteria identified in this study do not definitively define a neoplastic process, but appear to be helpful in individual cases. This study emphasises that endometrial changes should be always interpreted with the relevant clinical information, which would otherwise lead to overdiagnosis in premenopausal women.     

Postmenopausal bleeding of nonmalignant origin

A study was made of the medical records of 102 patients hospitalized because of postmenopausal bleeding. Diagnostic procedures used included vaginal examination, Papanicolaou smears, curettage and cervical biopsy. The major associated pathological conditions (possibly etiological factors) in the series were chronic cervicitis, fibromyoma of the uterus, endometrial polyps, cervical polyps and adenomyosis of the uterus. Sclerosis of the uterine vessels was suggested as another possible cause of this type of bleeding. Neither the amount and type of bleeding nor the pattern of associated symptoms were of diagnostic value.A history of hormonal therapy prior to the onset of bleeding is not sufficient evidence to establish that as the cause of the bleeding and the patient should be as completely investigated as if this history were not present. In over 61 per cent of cases in this series, uterine curettage with or without cervical biopsy, cauterization, conization or trachelorrhaphy, was the only treatment required for both diagnosis and therapy.     

Hormonal control of endometrial glycogen metabolism in the Macaca arctoides

Endometrial biopsies obtained throughout the menstrual cycle of the Macaca arctoides show the glycogen content paralleling the serum progesterone fluctuations which occur during the menstrual cycle. Secretory phase samples contained a three-fold higher concentration of glycogen when compared to follicular phase tissue. Changes in the activity levels of the glycogen metabolizing enzymes, glycogen phosphorylase and glycogen synthetase, during various stages of the menstrual cycle are in accord with the concept that the post-ovulatory increase in endometrial metabolism is a function of progesterone influence on this tissue. Endometrial glycogen synthetase activity remains low during the early proliferative phase of the cycle and becomes significantly elevated (two-to three-fold) during the early secretory phase of the cycle. Glycogen phosphorylase shows a similar cyclicity later in the luteal phase, reaching maximal activity between the seventeenth to nineteenth day of the cycle and remaining elevated through the twenty-sixth day of the cycle. The coincident nature of the rise in peripheral progesterone to increases in uterine glycogen metabolism suggest that progesterone may be the prime modulator of uterine endometrial metabolism during the post-ovulatory phase.     

In Vitro Effects of a Small-Molecule Antagonist of the Tcf/?-Catenin Complex on Endometrial and Endometriotic Cells of Patients with Endometriosis

Background

Our previous studies suggested that aberrant activation of Wnt/ß-catenin signaling might be involved in the pathophysiology of endometriosis. We hypothesized that inhibition of Wnt/ß-catenin signaling might result in inhibition of cell proliferation, migration, and/or invasion of endometrial and endometriotic epithelial and stromal cells of patients with endometriosis.

Objectives

The aim of the present study was to evaluate the effects of a small-molecule antagonist of the Tcf/ß-catenin complex (PKF 115–584) on cell proliferation, migration, and invasion of endometrial and endometriotic epithelial and stromal cells.

Methods

One hundred twenty-six patients (78 with and 48 without endometriosis) with normal menstrual cycles were recruited. In vitro effects of PKF 115–584 on cell proliferation, migration, and invasion and on the Tcf/ß-catenin target genes were evaluated in endometrial epithelial and stromal cells of patients with and without endometriosis, and in endometrial and endometriotic epithelial and stromal cells of the same patients.

Results

The inhibitory effects of PKF 115–584 on cell migration and invasion in endometrial epithelial and stromal cells of patients with endometriosis prepared from the menstrual phase were significantly higher than those of patients without endometriosis. Levels of total and active forms of MMP-9 were significantly higher in epithelial and stromal cells prepared from menstrual endometrium in patients with endometriosis compared to patients without endometriosis. Treatment with PKF 115–584 inhibited MMP-9 activity to undetectable levels in both menstrual endometrial epithelial and stromal cells of patients with endometriosis. The number of invasive cells was significantly higher in epithelial and stromal cells of endometriotic tissue compared with matched eutopic endometrium of the same patients. Treatment with PKF 115–584 decreased the number of invasive endometriotic epithelial cells by 73% and stromal cells by 75%.

Conclusions

The present findings demonstrated that cellular mechanisms known to be involved in endometriotic lesion development are inhibited by targeting the Wnt/β-catenin pathway.