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PubMed2003

Fetal cells in cervical mucus in the first trimester of pregnancy.

OBJECTIVES: The aim of this study was to first evaluate the presence of fetal cells in cervical mucus samples collected in the first trimester of pregnancy and then to compare different laboratory methods for the detection of these cells. METHODS: Mucus samples were collected by using a cytobrush before termination of pregnancy (TOP) from 143 pregnant women between 7 and 12 weeks of gestation. None of the women had undergone an invasive diagnostic procedure prior to cervical mucus sampling. Samples of placental tissue were collected from each patient at TOP. Slides from each sample were first observed under an inverted microscope to detect possible sperm contamination. In the first part of our experiments, 40 mucus samples were treated with a mucolytic solution containing N-acetylcysteine (AC) and were analysed by a polymerase chain reaction (PCR) assay. The second series, consisting of 71 mucus samples, was treated with a mucolytic solution containing dithiothreitol (DTT): all 71 samples were analysed by a PCR-based assay, and an aliquot for fluorescent in situ hybridisation (FISH) analysis was also obtained from 48 out of 71 samples. In the third part of our experiments, performed on 32 mucus samples, mucus trapped on the cytobrush was directly spread on two slides for FISH analysis without any mucolytic treatment. All placental tissue samples obtained at termination were analysed by FISH for fetal sexing. RESULTS: Overall, the use of PCR-based or FISH analyses on 143 mucus samples resulted in correct sex prediction in 92/143 (64.3%) samples [20/66 (30.3%) cases from known male pregnancies and 72/77 (93.5%) cases from known female pregnancies]. In the AC group, Y-derived sequences were found in 7/23 samples (30.4%) from known male pregnancies and in 1/17 cases from known female pregnancies, with an overall correct sex prediction in 23/40 cases (57.5%). In the DTT group, Y-derived sequences could be amplified in 10/30 samples (33.3%) from known male pregnancies and in 4/41 cases from known female pregnancies, with an overall correct sex prediction in 47/71 cases (66.2%). In the DTT samples analysed by FISH, nuclei bearing XY signals were detected in 5/26 (19.2%) cases from known male pregnancies and in none from female pregnancies, the rate of correct sex prediction being 56.2% (27/48). On untreated mucus samples analysed by FISH, nuclei with XY signals were documented in 3/13 (23%) samples from male conceptuses and in none from known female pregnancies, with an overall correct sex prediction in 22/32 cases (68.7%). CONCLUSION: Fetal cells were not detected in a constant and reliable fashion in cervical mucus samples collected in the first trimester of pregnancy. The detection rate was poorly influenced by the use of different laboratory methods. This sampling technique cannot be regarded as a promising tool towards minimally invasive prenatal diagnosis.

Adult↗
PubMed1992

Reversible contraception for the woman over 35 years of age.

Methods of reversible contraception, oral contraceptives, intrauterine devices, and Norplant (systemic progestin-only contraceptive; Wyeth-Ayerst, Radnor, PA), can be used for women over 35 years of age. Oral contraceptive formulations are safe and effective for healthy women up to the age of menopause. Oral contraceptives in women who do not smoke cigarettes do not result in a significant increased risk for cardiovascular disease. The incidence of breast cancer is not increased in women who have used oral contraceptives. A slight increase was found in younger women who had been on oral contraceptives based on a reanalysis of the contraceptive and steroid hormone study of the Centers for Disease Control. A reduction in the incidence of ovarian epithelial neoplasia by 40% was found in three European case-control studies. Two intrauterine devices are currently available on the US market: Paragard (GynoPharma, Somerville, NJ) and Progestasert (Alza Corp., Palo Alto, CA). Both of these provide highly effective contraception. A World Health Organization prospective randomized study found that there was an increase in pelvic inflammatory disease rates in the first 20 days after intrauterine device insertion. The intrauterine device itself did not increase the pelvic inflammatory disease incidence rates. The Norplant system exerts its contraceptive action through ovulation inhibition and alteration of cervical mucus. The major consumer complaint is irregular or prolonged uterine bleeding, which can be controlled by oral estrogen.

Adult↗
PubMed2004

Human cervical mucus can act in vitro as a selective barrier against spermatozoa carrying fragmented DNA and chromatin structural abnormalities.

PURPOSE: We have carried out experiments to determine if human cervical mucus can act as an in vitro selective barrier against spermatozoa morphologically normal that carry genetic structural abnormalities. METHODS: Sperm chromatin abnormalities have been evaluated by Chromomycin A3 and "endogenous" nick translation. RESULTS: The data obtained have shown that spermatozoa possessing higher levels of DNA protamination are more proficient in crossing the cervical mucus barrier. Moreover, the levels of positivity to endogenous nick translation treatment was practically zero in such spermatozoa. CONCLUSIONS: We suggest that sperm penetration of cervical mucus could be used to select sperm preparations free of fragmented DNA or chromatin structural abnormalities for assisted reproduction.

Cervix Mucus↗
PubMed1996

Reliability of trans-cervical recovery of placental cells from the lower uterine pole using a minimally invasive procedure. Evidence based on fetal sexing and analysis of recovered cell populations.

OBJECTIVE: Efficient recovery of placental cells (and their subsequent characterisation) from the lower uterine pole (L.U.P) by trans-cervical flushing or aspiration. SUBJECTS: Women attending for termination of pregnancy (7-17 weeks' gestation) for social reasons. All patients gave their consent to the procedures outlined below. METHODS: Trans-cervical intrauterine flushing (using 0.15 M NaCl) or mucus aspiration. Embryo transfer catheters were used in both procedures. Fetal sexing was achieved by gene amplification of Y-specific DNA sequences (Y-PCR), and by in situ hybridisation (bright-field and fluorescence) to the Y-chromosome. Data were compared with results obtained from fetal tissues recovered following termination of pregnancy. Gender-independent tests for fetal cells utilised immunocytochemistry with trophoblast-specific antibodies and dual immunocytochemistry/ISH, where appropriate. RESULTS: (1) Fetal sexing by Y-PCR: 71/122 (58%) aspirates contained Y-specific DNA. In addition, the sexing of 72/86 (84%) aspirates and their corresponding samples of placental tissue, agreed exactly. (2) Microscopic detection of fetal cells. Placentally-derived syncytiotrophoblast was detected in 17/45 (38%) flushings and 39/173 (23%) aspirates. In most other Y-PCR+ samples which were negative for syncytiotrophoblast, Y-chromosome-bearing nuclei of unknown origin, were observed by ISH and immunocytochemical evidence for cytotrophoblastic cells was also uncovered. CONCLUSIONS: Since Y-derived DNA can be detected in > 50% of flushings and aspirations, and gender-independent evidence for placental cells was obtained, regardless of fetal sex, we believe that most or all of these samples contained placental cells, including trophoblasts and naked nuclei. Trans-cervical placental cell recovery is a potentially valuable alternative to more invasive methods of aneuploid detection which require amniocentesis and CVS, provided its level of accuracy and above all, safety, can be evaluated.

Cervix Uteri↗
PubMed1979

The investigation of immunological infertility.

Immune reactions to spermatozoa may interfere with fertility in both males and females. These mechanisms may operate in a small, but significant, proportion of infertile couples, particularly when the duration of infertility is greater than three years. Antibodies to sperm may appear both in the blood and in genital tract secretions and their presence should be sought in both sites. The most suitable screening test for circulating anti-sperm antibodies in either partner is a method based on the complement-dependent antibody immobilization of sperm. This test may also be applied to cervical mucus as a routine in females. In males the detection of sperm immobilizing antibodies in blood serves as a reasonable guide to the existence of antibodies in seminal plasma where their presence may be confirmed using a gelatin agglutination test. Further information about the presence of locally secreted antibodies either in cervical mucus or in semen may be gained by using the sperm-cervical mucus contact (SCMC) test in which cross-testing with donor mucus or sperm will identify the site of local antibody production. Results based on immunofluorescence and sperm microagglutination methods are unreliable and lacking in correlation with continuing infertility and the routine use of these tests is not recommended.

Agglutination Tests↗
PubMed1967

Contraception.

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Cervix Mucus↗
PubMed1991

Assessment of the relevance of zona pellucida antibodies in serum and cervical mucus in patients who have fertilization failure during in vitro fertilization.

OBJECTIVE: To assess whether antibodies to the zona pellucida can cause fertilization failure during in vitro fertilization (IVF). DESIGN: Two methods for detecting zona pellucida antibodies were used to assess 20 women undergoing IVF. Nine patients had failed fertilization during the treatment cycle. SETTING: Academic IVF department of a teaching hospital. PATIENTS: Infertile patients previously assessed as requiring IVF. INTERVENTIONS: Gonadotropin-releasing hormone analogue to achieve pituitary down regulation and human menopausal gonadotropin for superovulation. Transvaginal ultrasound directed oocyte recovery. Cervical mucus and serum samples were obtained. MAIN OUTCOME MEASURES: The presence of antizona pellucida antibodies in serum or cervical mucus in relation to fertilization. RESULTS: Seven of 20 patients had antizona antibodies in serum and 7 of 20 patients had antibodies in mucus. Only 1 patient had antibodies in both samples. No correlation between the presence of antibodies in either sample and failure of fertilization was found. CONCLUSION: Detection of antizona pellucida antibodies by the methods described have no role in the prediction of IVF.

Antibodies↗
PubMed1988

Nitroblue tetrazolium reduction by leukocytes in the cervix of pregnant women.

We examined nitroblue tetrazolium reduction activity in leukocytes from cervical mucus as well as peripheral blood from pregnant and nonpregnant women by histochemical methods. More than 90% of the cells examined were polymorphonuclear cells. Nitroblue tetrazolium reduction in polymorphonuclear cells from peripheral venous blood was significantly enhanced in pregnant women compared with that in nonpregnant women. However, there was no difference in activity between pregnancy periods. On the other hand, nitroblue tetrazolium reduction in cervical polymorphonuclear cells excluding women with cervicitis was also greater in pregnant women than in nonpregnant women. Moreover, activity in cervical polymorphonuclear cells gradually increased as pregnancy progressed. This enhanced activity in polymorphonuclear cells may be involved not only in the defense mechanism but also in cervical ripening in pregnancy.

Cervix Mucus↗
PubMed1988

The influence of copper intrauterine device on fatty acid composition of cervical mucus lecithin.

Fatty acid composition of lecithin was analysed in midcycle cervical mucus obtained from 19 women using a copper intrauterine device (Cu-IUD) and 16 controls. In both groups, palmitic (16:0) acid was the predominant fatty acid and oleic (18:1) acid the second largest component. In women using a Cu-IUD, levels of myristic (14:0) acid were significantly lower, while levels of linoleic (18:2) and docosahexaenoic (22:6) acid were significantly higher than in controls. These changes in fatty acid composition could not be related to duration of IUD use and were very similar to those observed in women with primary unexplained infertility, suggesting that the Cu-IUD-induced modification of fatty acid pattern in midcycle mucus lecithin may be part of the contraceptive mechanism of action.

Adolescent↗
PubMed1989

Pharmacodynamics of a contraceptive vaginal ring releasing 3-keto-desogestrel.

The pharmacodynamic effects of a new type of 3-keto-Desogestrel [3kDOG] releasing vaginal ring was studied in a group of 20 normally ovulating women during a period of 21 days continuous use. Peripheral blood samples were taken for the estimation of progesterone and oestradiol levels, ultrasound tracking for follicular growth and cervical mucus sampling for scoring [modified Insler] and sperm penetration testing during a control, treatment and recovery period. Additional blood samples were taken for the estimation of 3kDOG levels during the treatment period. After a control cycle, twenty normally ovulating women were selected and randomly divided into two groups. Group A were given a vaginal ring on day 5 of the menstrual cycle releasing 30 ug/24 hours of 3kDOG and group B a similar ring releasing 15 ug/24 hours of kDOG. Of the treatment cycles, none showed an ovulatory pattern in group A and there was only one in group B, the respective mean plasma levels of 3kDOG were 0.38 and 0.25 pmol/ml. The formation of "cysts" or persistent follicles was common to both groups, 6 out of 10 subjects in group A and 8 out of 10 subjects in group B had "cysts" greater than 25mm in diameter in the treatment cycle, this had become 5 out of 10 and 8 out of 10, respectively, in the recovery cycle. A comparison of the cervical mucus scores and sperm penetration of cervical mucus showed a significant reduction between the control and treatment groups but no significant difference between the two groups. The bleeding profiles showed an increase in percentage of bleeding days in both treatment groups to 25% in group A and 21.7% in group B [cf, 17.1%, 15.9%, respectively, in control month]; there being no statistical difference between groups. In conclusion, it is believed that given the efficacy demonstrated and the low level of menstrual disturbances found, this system warrants further investigation as a means of contraception as it appears to offer a better compromise than previous vaginal ring systems.

Administration, Intravaginal↗
PubMed1996

Symmetry and ovulation in women.

Women have cryptic ovulation, and self-observation methods of determining the timing of ovulation, such as monitoring cervical mucus symptoms or recording basal body temperature, are not reliable. It has recently become apparent that the symmetry of paired soft tissue traits, such as breast size and digit length, changes across the menstrual cycle. This paper presents evidence that symmetry in four paired soft tissue traits showed a marked increase on the day of ovulation. The difference (i.e. the asymmetry) between the size of the left and right trait in ears, 3rd, 4th and 5th digits was measured. The timing of ovulation was confirmed by real-time pelvic ultrasonography and trait measurements were made without knowledge of scan results. Asymmetry was lowest on the day of ovulation (day 0), decreasing by about 30% from day -1, and significant within-subjects changes occurred from days -2 to day -1, and day -1 to day 0. The practical and evolutionary implications of these findings are discussed.

Adult↗
PubMed1992

Infertility in women: diagnostic evaluation with hysterosalpingography and other imaging techniques.

This article examines the current roles of hysterosalpingography and other imaging studies used in diagnostic assessment and treatment of infertility in women. The advantages and limitations of hysterosalpingography, transabdominal or transvaginal sonography, and MR imaging in detecting abnormalities of the uterus (congenital anomalies, intrauterine adhesions, myometrial and endometrial derangements) and tubal and peritubal disease are presented. The use of sonography to evaluate cyclical endometrial changes, to assess development of the follicle and corpus luteum, to determine whether ovulation has occurred, and to detect dysfunctional ovulatory dynamics is described, as is the role of this technique in improving the prognostic value of the postcoital test for assessing the quality of cervical mucus. With the development and availability of newer and improved methods for the treatment of infertility disorders in women, comprehensive and detailed evaluation to determine the precise underlying cause or causes of the infertility is important to allow formulation of the most appropriate therapeutic plan. With careful and thorough assessment, the cause of infertility can be identified in 90-95% of couples evaluated.

Adult↗