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Electrolyte interactions in cervical mucus and their relationship to circulating hormone levels.

The changes in electrolyte composition of cervical mucus (CM) were monitored during the menstrual cycle, using scanning electronic microscopy (SEM) and energy dispersive X-ray analysis (EDX). Correlations were sought between the electrolytes present in CM, the physical characterisitics of the CM, and circulating levels of gonadal steroids and protein hormones. The results suggest that there is a definite relationship between circulating estrogen levels and electrolyte composition of CM, and also provide information on the secretion of electrolytes during the human menstrual cycle. This information will be of use in development of methods for fertility control directed to the uterine cervix.

Adolescent↗

[Effect of successive ejaculation on semen analysis parameters in normal men].

OBJECTIVE: To study the effect of successive ejaculation on semen analysis parameters in normal men. METHODS: Eight ejaculates were collected at daily intervals from 8 normal men. The semen parameters were analyzed according to WHO laboratory manual for the examination of human semen and sperm-cervical mucus interaction. RESULTS: The semen volumes and total sperm counts decreased gradually day by day, significantly lower from the 5th to the 8th day than the 1st (P < 0.01). The sperm densities also declined day by day, but not significantly. The sperm viability and motility (a + b grade) presented an increasing trend, but with no significance except on the 7th day. CONCLUSION: With the increase of the ejaculatory frequency, human semen volumes and the total counts significantly decrease, while sperm viability and motility gradually increase. Successive ejaculation does not affect the quality of human semen.

Adult↗

Differential expression of MUC genes in endometrial and cervical tissues and tumors.

BACKGROUND: Mucin glycoprotein's are major components of mucus and are considered an important class of tumor associated antigens. The objective of this study was to investigate the expression of human MUC genes (MUC1, MUC2, MUC5B, MUC5AC and MUC8) in human endometrium and cervix, and to compare and quantitate the expression of MUC genes in normal and cancerous tissues. METHODS: Slot blot techniques were used to study the MUC gene expression and quantitation. RESULTS: Of the five-mucin genes studied, MUC1, MUC5B and MUC8 showed high expression levels in the normal and cancerous endometrial and cervical tissues, MUC2 and MUC5AC showed considerably lower expression. Statistically, higher levels of MUC1, MUC5B and MUC8 were observed in endometrial adenocarcinomas compared to normal tissues. In contrast, only MUC1 levels increased with no significant changes in expression of MUC5B and MUC8 in cervical tumors over normal cervical tissues. CONCLUSION: Endometrial tumors showed increased expression of MUC1, MUC5B and MUC8 over normal tissues. Only MUC1 appears to be increase, in cervical tumors. All the studied tissues showed high and consistent expression of MUC8 mRNA. Low to neglible levels of MUC2 and MUC5AC were observed in all studied endometrial and cervical tissues.

Adenocarcinoma↗

Ovarian function during use of a levonorgestrel-releasing IUD.

Ovarian function was studied for two complete menstrual cycles in 9 regularly menstruating women and for 8 weeks in three amenorrhoeic women who had used levonorgestrel-releasing IUDs (LNG-IUD) for more than four years. Nine patients using copper IUDs (Nova-T) were studied for two complete menstrual cycles as controls. According to progesterone levels, 15/17 cycles in women using LNG-IUDs were ovulatory, whereas only 8/17 cycles showed normal follicular growth and rupture as judged by ultrasound. In ovulatory cycles, the peak progesterone levels were lower than in the controls. The preovulatory estradiol and LH peak levels were also lower than in control subjects. SHBG levels were lower in LNG-IUD users than in copper IUD users. It is concluded that, although the dose of levonorgestrel released from the IUD is very low, it probably exerts an effect on the gonadotrophin secretion, which disturbs follicular development in many of the women studied, which in addition to the local effect on the endometrium, contributes to its high contraceptive efficacy.

Adult↗

Localization of binding sites of naturally occurring antisperm antibodies on human spermatozoa by immunofluorescence.

Antisperm antibodies (ASA) may affect sperm motility, acrosome reaction, sperm penetration of cervical mucus, binding to the zona pellucida, and sperm-egg fusion. We investigated the localization of ASA of infertile men or men after vasectomy bound on the sperm surface using an immunofluorescence method. Binding occurred in the acrosomal region, midpiece, and tail. Most of the ASA in both groups of patients bound to the midpiece alone or in combination with other regions of spermatozoa. Only few ASA samples showed binding to all the three sperm regions. A combination of binding to the acrosomal region and to the midpiece was never observed. In infertile patients with ASA, the binding site was compared with sperm parameters. ASA binding to the sperm head influenced the acrosome reaction. Binding of ASA on tail and/or midpiece was not associated with a significant alteration of viability and motility. Immunofluorescence appears to be a valuable tool in the diagnosis of immune infertility, in particular when impairment of the acrosome activity is suggested.

Acrosome↗

The leukocytic reaction of the human uterine cervix.

PROBLEM: Sperm interaction with the immune system of the human cervix is poorly understood. METHOD: The leukocytic response of the human cervix to sperm was examined in a closely monitored patient population (N = 10), using monoclonal antibody cell identification techniques. Baseline data were collected from both cervical mucus and smears sampled before treatment by donor insemination. Donor insemination was timed to coincide with ovulation by monitoring plasma LH concentrations twice daily. Following insemination the numbers of leukocytes were recorded in cervical mucus and smear samples taken over a 24-h period relative to the time of treatment. Controls treated with "pure sperm," seminal plasma, cryopreservative, and cervical smearing alone were also included in the study. RESULTS: Only those women treated with sperm cells exhibited substantial elevations in leukocyte numbers following inseminations. Additionally, serial cervical smearing induced an inflammatory response of the cervix. In all the women, the neutrophil was the predominant leukocyte of the cervix both during the baseline and treatment periods (median values ranged from 77 to 86%). Macrophages, T-helper lymphocytes, and T-suppressor lymphocytes were also detected, but only in low numbers (2-10.6%). Two patients and one control ("pure sperm") became pregnant during their study cycle. CONCLUSIONS: We conclude that the leukocytic reaction is a physiological response of the cervix to sperm, the function of which remains to be established.

Cell Movement↗

[Good rules for cervical cytologic sampling].

There are still many little problems concerning methods about cervical cytology sampling. Are there some risks of teaching Papanicolaou smear technique? In which day of the cycle is it better to do sampling? How long in advance is it better to avoid sexual relations, and do not use any vaginal douching? Is it better to remove in advance cervical mucus? When should the sampling be done if the colposcopy is made in the same time? How should you behave if the cervix is not exposed? How many slides have to be spreaded? Which is better, the fixation with alcohol or aerosol? Where should the sampling be repeated? Is the patient anamnesis reliable? What should it mean the presence of some parasites and some foreign bodies manifested on the slide? Different answers have been given to reply to these questions and to some others. Smear's collecting is a simple method, but it needs an exact manual skill and an adherence to some precise rules. A relevant part of the final result depends how sampling has be done. Improvements must be done in every link of chain events.

Colposcopy↗

Synthetic cervical mucus formulation.

A synthetic formulation has been developed with viscosity, spinnbarkeit, and pH comparable to that reported for human cervical mucus. The formulation contains guar gum crosslinked with borate ion, mucin (dried porcine gastric), and a mixed preservative system in pH 7.4, 0.1M phosphate buffer. The guar gum source, mucin concentration, and method of preparation were shown to be critical factors in the performance of the formulation.

Borates↗

Treatment of sperm autoimmunity in men.

Positive sperm immobilization tests were found in the sera of 6.3% of 491 infertile men. There was a higher frequency of family histories of autoimmune diseases than for other subfertile men. Prognosis for fertility was related to results of sperm-mucus penetration tests: progressively motile sperm indicated a fair outlook--8 of 15 couples conceived within 7 months either spontaneously or with artificial insemination with husband's semen. Absence of progressively motile sperm indicated severe infertility - only 6 of 72 couples produced pregnancies. Treatment of the latter group by temporary suppression of spermatogenesis with testosterone or with Artificial Insemination by Husband (AIH) appeared to be ineffective. However, sperm immobilization titres fell and sperm concentrations, motility and mucus penetration increased during treatment of 14 of the men with prednisolone (0.5-0.75 mg/kg per day) for 4-7 months and there were three pregnancies. It is concluded that sperm autoimmunity in men causing failure of sperm to penetrate cervical mucus produces severe but not absolute infertility. Immunosuppression with continuous high dose glucocorticoid reduces antibody levels and improves semen quality. A controlled trial is needed to determine if this treatment is a practical method for improving fertility in men with sperm autoimmunity.

Adult↗

[Correlations between penetration tests and spermatozoal antibodies (author's transl)].

The results obtained from penetration testing of cervical mucus of 60 sterile couples are reported in this paper. Also, the presence of spermatozoal antibodies in the serum of both the female and male probands was detected by means of the immunofluorescence method. --Neither in the females nor males was any relationship detectable between the occurrence of spermatozoal antibodies and the results of penetration testing. Yet, spermatozoal antibodies were found to occur much more often in probands with sterility of unelucidated origin.

Antibodies↗

Detection of mutagens in cervical mucus in smokers and nonsmokers.

The Salmonella mutagenicity test was used to analyze cervical mucus specimens from 364 smokers and 333 nonsmokers to determine whether the association between smoking and mutagenic cervical mucus that we reported previously among women diagnosed with dysplasia would apply to a larger group of healthy women (E. A. Holly et al., J. Natl. Cancer Inst., 76: 983-986, 1986). Women smokers and nonsmokers between the ages of 18 and 49 who attended eleven clinics and physicians' offices in the San Francisco Bay area for a routine Pap smear were examined to determine whether smokers were more likely to have mutagenic substances in their cervical mucus. About 4% of smokers and 8% of nonsmokers had positive mutagenicity test results (P = 0.02). Cervical mucus with a large number of microorganisms was more likely to have a positive mutagenicity test result than that with fewer microorganisms (test for trend, P = 0.01). Mutagenicity results varied by race and clinic location but were not associated with smoking behavior, sexual behavior, gynecological diagnosis, or diet. Further work is needed to develop methods to detect mutagens in specific body fluids.

Adolescent↗

Six kinds of anti-phospholipid antibodies (aPLs) in ovulatory mucus and seminal plasma from couples with repeated miscarriages.

OBJECTIVE: Determination of aPLs in ovulatory mucus and in seminal plasma in 18 infertile couples (women, median age 33.2, range 28-39 years; men, median age 36.4, range 29-46 years) with repeated unexplained miscarriages (four to nine spontaneous abortions). Women with high levels of aPLs in sera were excluded from the study. METHODS: aPL screening included cardiolipin (Sevac Prague), L-alpha-phosphatidic acid, L-alpha-phosphatidylethanolamine, L-alpha-phosphatidyl-DL-glycerol, L-alpha-phosphatidylinositol, and L-alpha-phosphatidyl/serine (Sigma, USA). These were determined in isotypes IgG and IgA by an ELISA method. Statistical analysis of aPL results was made by cut-off levels of aPLs for both Ig isotypes by using 3 SD or 95th percentile calculated by STATGRAPHICS. Control group was created from four fertile healthy couples. RESULTS: Levels of aPLs in ovulatory cervical mucus were in the majority very low, except for IgA-aPLs-inositol. Very high interindividual differences in aPLs were found in seminal plasma. IgA-aPL-ethanolamine and -DL-glycerol, and IgG-aPL-L-serine and -DL-glycerol predominated in seminal plasma. Extremely high IgA levels of aPLs in all above-mentioned antigens, except cardiolipin, were found in one man of an infertile couple. CONCLUSION: The passage of abnormal aPLs levels from seminal plasma into the site of conception during often repeated, unprotected intercourses is supported by the findings.

Abortion, Habitual↗

Efficacy of methods for determining ovulation in a natural family planning program.

OBJECTIVE: To evaluate the efficacy in ovulation detection of methods used in natural family planning in comparison with pelvic ultrasonography. DESIGN: Prospective analysis of ovulation detection by natural family planning methods and ultrasonography. SETTING: Natural family planning clinic, Department of Obstetrics and Gynecology, University of Naples "Federico II". PATIENT(S): Forty healthy women who were highly motivated to use natural family planning. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Transvaginal ultrasonographic findings, urinary LH levels, salivary beta-glucuronidase activity, salivary ferning levels and characteristics of cervical mucus, and BBT. RESULT(S): Urinary LH level determination yielded a 100% correlation with the simultaneous ultrasonographic diagnosis of ovulation. Mucus sensations and characteristics yielded a 48.3% correlation when simultaneously evaluated with ovulation. Beta-glucuronidase levels yielded a 27.7% correlation. The salivary ferning test had a 36.8% ovulation-detection rate the day of ovulation, but 58.7% of results were uninterpretable. Body temperature measurements yielded a 30.4% correlation with the simultaneous ultrasonographic diagnosis of ovulation. CONCLUSION(S): Measuring urinary LH levels is an excellent method for determining ovulation. Although variations in mucus characteristics and basal body temperature correlate somewhat with ovulation, the length of the fertile period is overestimated with these methods. The salivary ferning test and measurement of beta-glucuronidase levels are not good methods for home ovulation testing.

Adult↗

Levonorgestrel levels in serum and uterine flushings in women bearing LNG-Cu-IUD.

Radioimmunoassay was employed to measure levonorgestrel levels in serum and uterine flushings in women using intrauterine device releasing both copper ions and levonorgestrel (LNG-Cu-IUD) after one-year insertion. In the meanwhile physical characteristics of cervical mucus have been observed in women using LNG-Cu-IUD and compared with those using Tcu-220c-IUD. The mean concentrations of levonorgestrel in serum and uterine flushings (5 ml) were 84.53 pg/ml and 266.93 pg/ml respectively. In LNG-Cu-IUD group, the cervical mucus lacking in ferny crystals had a viscosity significantly higher than that in Tcu-220c-IUD group, suggesting that the local high concentration of levonorgestrel changed the environment of the uterine cavity and the cervical mucus status.

Adult↗

Temporal relationship between Uniplant insertion and changes in cervical mucus.

This study was undertaken to determine the time required by a single implant containing nomegestrol acetate to affect cervical mucus production and sperm penetration in women. All subjects were investigated and, if necessary, treated for any kind of cervicitis or vaginitis prior to starting cervical mucus study. The subjects had not used hormonal contraception for at least three months prior to investigation. They were counseled to use condoms during this study and also to refrain from intercourse during the period of cervical mucus sampling. Follicular development and endometrial thickness were analyzed by transvaginal sonography. Cervical mucus examination, sperm penetration test, and transvaginal sonography were performed during the control cycle and during the first cycle of Uniplant use. Blood samples were taken for the measurement of estradiol, LH, and progesterone. Cervical mucus and sperm penetration tests were evaluated according to the World Health Organization (WHO) criteria. In the treated cycle, when cervical mucus reached a score of 8-10, Uniplant was inserted, independent of the day of the cycle. Cervical mucus was then collected at 0, 4, 8, 12, 24, 48, and 96 h later until a marked change in volume, consistency, ferning spinnbarkheit, and cellularity was observed. All samples were also used for sperm penetration test. Preovulatory estradiol and LH peak decreased significantly compared to pre-implant insertion. Progesterone levels were within the normal limit. Cervical mucus and sperm penetration tests were not affected by Uniplant in the first 12 h. Twenty-four hours after Uniplant insertion, cervical mucus and sperm penetration tests were affected in 70.6% of the women. Forty-eight hours after implant insertion, the women were affected. Follicular rupture occurred in the majority of the women 48 h after implant insertion. Based on these results, it is possible to conclude that Uniplant can affect estradiol and LH preovulatory peaks and disrupt the process of cervical mucus production and sperm penetration, but it was unable to prevent ovulation when inserted in the preovulatory phase.

Adult↗

A phase I comparative study of contraceptive vaginal films containing benzalkonium chloride and nonoxynol-9. Postcoital testing and colposcopy.

Benzalkonium chloride (BZK) has been shown in in vitro and in vivo studies to immobilize sperm, to be active against STD-causing organisms, and to penetrate and thicken cervical mucus. A US clinical study of a sponge containing 60 mg BZK showed life table pregnancy rates at 6 and 12 months of 11.7 and 18.9 per 100 women, respectively. BZK is not currently marketed in the US as a contraceptive. The present study aimed to assess 1) the ability of a new contraceptive vaginal film containing either of two doses of BZK to prevent the penetration of sperm into midcycle cervical mucus; 2) the effect of the film on the vaginal epithelium; and 3) the acceptability of the film. All results were compared with VCF, a currently marketed film containing nonoxynol-9 (N-9). Ten women underwent two baseline cycles of postcoital testing in which no film was used, followed by three test cycles in which Allendale-BZK film, a new film containing either 19 or 25 mg BZK, or VCF, containing 70 mg N-9, was used. The sequence of testing cycles was randomized. In each cycle, condoms were used prior to midcycle, then a midcycle cervical mucus specimen was examined to ensure midcycle characteristics and the absence of sperm. Each woman then had intercourse using either no film (baseline cycles) or a test film (test cycles) and returned 2-3 h afterwards. Cervical mucus was again assessed for adequacy and the presence of sperm. Each woman also underwent colposcopy, using a protocol developed by the World Health Organization. The average number of progressively motile sperm seen per high power field was as follows: first baseline cycle, 22.2; second baseline cycle, 22.1; test cycle with film containing 19 mg BZK, 0.2; test cycle with film containing 25 mg BZK, 0.0; and test cycle with VCF containing 70 mg N-9, 0.0. There was no significant difference between baseline cycles or among test cycles in the average number of progressively motile sperm seen (p = 0.78 and p > or = 0.75, respectively). The average number of progressively motile sperm seen in each test cycle did, however, differ significantly from the average number seen in either baseline cycle (p < 0.01). Colposcopy showed superficial de-epithelialization without underlying inflammation in 15-20% of baseline cycles, regardless of whether colposcopy was done before or after coitus; in 50% of cycles in which either dose of BZK was used; and in 69% of cycles in which VCF was used. In all cases women were asymptomatic. Erythema and petechiae were also seen on colposcopy although at a lower frequency than de-epithelialization. There was no difference in the acceptability of the films. A vaginal contraceptive containing either 19 or 25 mg BZK in a new film base appears to be comparable with VCF in preventing sperm from entering midcycle cervical mucus and may be somewhat less disruptive to the vaginal epithelium.

Administration, Intravaginal↗