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Preclinical evaluation of sodium cellulose sulfate (Ushercell) as a contraceptive antimicrobial agent.

The spread of sexually transmitted infections (STIs) and limited methods for control of pregnancies presents high risks to the reproductive health of women. Methods controlled by women and directed toward disease prevention and contraception are needed. We report on preclinical studies of the biological properties of sodium cellulose sulfate (Ushercell) currently being developed for use as a topical contraceptive antimicrobial agent. Ushercell was evaluated with tests designed to identify its contraceptive and antimicrobial properties. Ushercell inhibits hyaluronidase (reversible; IC50 = 1.7 mg/mL), impairs sperm penetration of cervical mucus (approximately 70% inhibition at 1 mg/mL), and acts as a stimulus for acrosomal loss (IC50 = 52 ng/mL). It prevents conception in rabbits when added to spermatozoa (approximately 95% inhibition at 1 mg/mL) or when vaginally applied (complete contraception by 45 mg) before insemination. However, up to 50 mg/mL, Ushercell does not irreversibly immobilize spermatozoa, suggesting that Ushercell is not cytotoxic. Ushercell has a broad spectrum of antimicrobial activity in vitro. Inhibited microbes include human immunodeficiency viruses (different laboratory strains and clinical isolates; IC50 values range from 3 to 78 microg/mL), herpes viruses, HSV-1 (IC50 = 59 ng/mL) and HSV-2 (lC50 = 24 ng/mL), Neisseria gonorrhoeae (IC50 = 2 microg/mL), and Chlamydia trachomatis (IC50 = 78 microg/mL). In contrast, Ushercell does not inhibit growth of beneficial vaginal bacteria, Lactobacillus gasseri, at 5 mg/mL. These results suggest that the antimicrobial effects of Ushercell are selective, and not likely mediated by nonspecific cytotoxic mechanisms. These data provide the basis for further clinical development of Ushercell as a vaginal agent to prevent unplanned pregnancy and STIs.

Acrosome↗

Cervicectomy with uterovaginal anastomosis in the rhesus monkey: a new surgical model for reproductive studies.

Cervicectomy with uterovaginal anastomosis in the rhesus monkey allows easy and non-traumatic access to the uterine cavity through the vagina for such purposes as biopsy, flushing, aspiration of fluid, and placement of IUDs. Endometrial and tissue samples cannot be contaminated by cervical mucus. The surgical procedure and results on eleven animals are described. Neither uterine nor ovarian function appeared to be significantly altered. The surgical model seems well suited to long-term studies on IUDs.

Animals↗

Effects of the anti-oestrogens, clomiphene and tamoxifen, on the cervical factor in female infertility.

The anti-oestrogens clomiphene and tamoxifen are successful therapeutic agents in the induction of ovulation but the associated pregnancy rates are low. One cause for this may be a direct anti-oestrogenic effect on cervical mucus, reducing its sperm penetrability. This potentially adverse effect has been shown to occur with clomiphene. In this study the effects of clomiphene and tamoxifen on the cervical factor are compared in women who were anovulatory or had defective ovulation. Forty fully investigated infertile women were randomly allocated to different treatment sequences after being observed for three control cycles. A double-blind placebo technique was used with four different treatment regimes. Serial serum progesterone and oestradiol were assayed. Cervical mucus from the periovulatory phase was assessed by means of the cervical score and in vitro sperm penetration tests. Both these anti-oestrogens can affect cervical mucus but their effect seems to be related more to the end-organ sensitivity of the individual than to a specific action of the drug per se.

Cervix Mucus↗

The reproducibility of the postcoital test: a prospective study.

OBJECTIVE: To determine the reproducibility of the postcoital test among trained observers. METHODS: Twenty-eight infertile patients presenting to the Brigham and Women's Hospital over a 1-year period were recruited for the study. After a standardized collection of specimens for the postcoital test, four fellowship-trained reproductive endocrinologists evaluated six postcoital test characteristics and gave their overall impression of the test. Each observer was blinded to the patients' identities and clinical histories as well as to the ratings of the other observers. The six characteristics included an assessment of the cervical mucus by ferning, cellularity, spinnbarkeit, and consistency, and of sperm by total count per high power field and percent motility. Scoring was adapted from World Health Organization (WHO) criteria for semen-cervical mucus interaction. Statistical analysis included the kappa statistic to determine agreement among observers for postcoital test characteristics and the Mantel-Haenszel test to determine the association between overall impression and the other test characteristics. RESULTS: Agreement among the four observers was best for sperm number and motility (39% of cases) and worst for cellularity, spinnbarkeit, and overall test impression (11, 14, and 14% of cases, respectively). The kappa statistic ranged from a low of 0.13 for cellularity, demonstrating poor reliability (95% confidence interval [CI] 0.03-0.23), to a high of 0.51 for sperm number, demonstrating fair reliability (95% CI 0.41-0.60). Only sperm number and percent motility were significantly associated with the overall impression (P < .001). CONCLUSIONS: In a blinded study, the characteristics of the postcoital test were found to have poor to fair reproducibility among trained observers using a standardized WHO scoring system. The observers' overall impressions of test quality correlated with sperm number and motility only. We question the validity of the postcoital test as a diagnostic tool in the evaluation of infertility.

Adult↗

[Detection of anti-sperm antibodies on the surface of live spermatozoa by the method of flow cytometry].

Detection of antisperm antibodies is an important step of the diagnosis of infertility. In present study we conducted comparative studies with the detection of antisperm antibodies in IgG, IgA and IgM classes on the surface of living spermatozoa by flow cytometry and with Mixed Agglutination Reaction and Sperm immobilization. Our data show that flow cytometry may be used to verify that immunoglobulins are on the surface of living spermatozoa, either retrieved directly from an ejaculate or following exposure to cervical mucus or serum and to determine their isotype, proportion of sperm which bound antibody and quantity of immunoglobulins bound to cell surface.

Animals↗

New approaches to fertility awareness-based methods: incorporating the Standard Days and TwoDay Methods into practice.

Helping clients select and use appropriate family planning methods is a basic component of midwifery care. Many women prefer nonhormonal, nondevice methods, and may be interested in methods that involve understanding their natural fertility. Two new fertility awareness-based methods, the Standard Days Method and the TwoDay Method, meet the need for effective, easy-to-provide, easy-to-use approaches. The Standard Days Method is appropriate for women with most menstrual cycles between 26 and 32 days long. Women using this method are taught to avoid unprotected intercourse on potentially fertile days 8 through 19 of their cycles to prevent pregnancy. They use CycleBeads, a color-coded string of beads representing the menstrual cycle, to monitor their cycle days and cycle lengths. The Standard Days Method is more than 95% effective with correct use. The TwoDay Method is based on the presence or absence of cervical secretions to identify fertile days. To use this method, women are taught to note everyday whether they have secretions. If they had secretions on the current day or the previous day, they consider themselves fertile. The TwoDay Method is 96% effective with correct use. Both methods fit well into midwifery practice.

Adult↗

Echographic study of follicular maturation and ovulation during the normal menstrual cycle.

Follicular development was followed by echography during 18 menstrual cycles in 10 women volunteers. Results were correlated with changes in serum hormone levels, basal body temperature curves, and cervical mucus changes. The average follicular growth as measured from the time of echographic visualization was 0.3 cm/day. The diameter of the follicle measured the day before disappearance was 2.7 +/- 0.3 cm. In all cases echography was shown to indicate accurately the time of ovulation. Given its inoffensive nature, the usefulness of echography in determining the physical event of ovulation is demonstrated. Possible applications in the surveillance of ovulation induction methods and in artificial insemination are discussed.

Adult↗

[Immunological indices in differential diagnosis of active and inactive chronic endocervicitis].

A total of 132 patients were examined for chronical endocervicitis by clinical, immunological and morphological methods. According to the results obtained the patients were subdivided into several groups. The first group covered 77 females with an active clinical somatic form of the disease. The second group included 55 patients with chronical inactive endocervicitis. Controls included 25 practically healthy females. No differences were found between immunological indices of peripheral blood in groups of patients with active and inactive forms of chronic endocervicitis. Immunological tests on cervical mucus in patients with chronic endocervicitis revealed differences in the total number of leukocytes, content of living cells, functional activity of neutrophils, concentration of immunoglobulins and proinflammatory cytokines. This study may contribute to the development of immunological criteria for diagnosis of inflammatory process activity as well as a diagnostic model.

Carrier State↗

Phase I comparative clinical trial with subdermal implants--bioabsorbable levonorgestrel or norethisterone pellet fused with cholesterol.

The potential for antifertility effect of two bioabsorable pellets, one containing norethisterone (NET) and the other containing levonorgestrel (LNG) fused with cholesterol, was studied in a group of healthy, menstruating but sterilised women. The pellets weighed approximately 30 mg and contain 85% steroid and 15% cholesterol. A single NET pellet was inserted in 4 subjects, out of which 2 were also studied for steroid pattern in blood. After reaching peak levels within 48 hours, the plasma NET levels declined gradually within a fortnight's time, and thereafter, ranged between 200 to 700 mg/ml up to 90 days post-insertion. After this period, occassional spurts of NET release were seen. Bleeding pattern was studied in 37 cycles; mid-luteal progesterone (P) estimation was done in 16 cycles, mid-cycle cervical mucus was studied in 27 cycles and post-coital test (PCT) in 7 cycles. Cycle length with pellet insertion was of 25 to 37 days duration except one cycle of 55 days duration. All cycles studied during the treatment were ovulatory (P greater than 5 ng/ml) and no consistent effect was observed on cervical mucus as well as on PCT. Thus, a single NET pellet does not seem to have a reliable contraceptive potential. A single LNG pellet was inserted in 8 subjects, and steroid patterns were studied in 6 women. Peak levels of LNG were reached within 24 hours of pellet insertion and the levels fell gradually in a week's time. Thereafter, LNG levels ranged between 100-400 pg/ml up to 8 months. In contrast to the observation with NET pellets, only one subject showed frequent spurts of LNG release until the 4th month of treatment. Bleeding pattern was studied in 90 cycles, P was estimated in 52 cycles, mid-cycle cervical mucus studied in 55 cycles and PCT done in 26 cycles. Breakthrough bleeding (BTB) occurred on 15 occasions, 11 episodes being in 2 subjects. Cycle length varied between 21 to 42 days. The LNG pellet did not exert a consistent effect either on ovulation inhibition or cervical mucus or PCT. In their present form, therefore, a single LNG pellet also does not appear to have a reliable contraceptive effect.

Absorption↗

Sperm immobilizing antibodies interfere with sperm migration from the uterine cavity through the fallopian tubes.

PROBLEM: It is well known that sperm migration in cervical mucus is impaired by sperm immobilizing antibodies secreted in the mucus. However, it is not clear yet whether sperm migration from the uterine cavity through the fallopian tubes to the peritoneal cavity is impaired by sperm immobilizing antibodies. To test the possible impairment of sperm migration in the tubes, laparoscopic examinations were carried out and the presence of motile sperm in the peritoneal fluid after intra-uterine insemination was investigated. METHOD: Peritoneal sperm recovery tests were performed in 28 infertile women with sperm immobilizing antibodies in their sera, and the results were compared with those in 322 infertile women without the antibodies. Both the sperm immobilizing antibody titers (SI50) and complement activities (C'H50) in peritoneal fluid were compared with those in patients' sera. In some experiments, the supernatant of the peritoneal fluid was used as a source of complement for the sperm immobilization tests instead of guinea pig serum. RESULTS: Among couples with normal semen characteristics by the criteria of WHO, sperm recovery in the peritoneal fluid was observed in only 3 (11.1%) of 27 patients with sperm immobilizing antibodies, compared with 72 (34.0%) of 212 patients without the antibodies (P < 0.025). The antibody titers of the patients with the sperm recovery were very low by the quantitative sperm immobilization test. In most patients, a similar amount of sperm immobilizing antibodies was present in the peritoneal fluid and the sera. Though the complement activities in the peritoneal fluid were less than those in sera, the former were still found to be sufficient to immobilize sperm in vivo. CONCLUSIONS: These results suggest that the complement-dependent sperm immobilizing antibodies could interfere with sperm migration in the female genital tract at the level of the fallopian tubes.

Animals↗

Interaction of St. John's Wort with oral contraceptives: effects on the pharmacokinetics of norethindrone and ethinyl estradiol, ovarian activity and breakthrough bleeding.

OBJECTIVES: This study evaluated the effect of the herbal remedy St. John's Wort on oral contraceptive (OC) therapy with respect to the pharmacokinetics of norethindrone and ethinyl estradiol, ovarian activity and breakthrough bleeding. METHODS: Sixteen healthy women were treated with a low-dose OC (Loestrin 1/20) and a placebo for two consecutive 28-day cycles in a single-blind sequential trial. Treatment with St John's Wort 300 mg three times daily was then added for two additional 28-day cycles. Outcomes compared between control and treatment cycles included the pharmacokinetics of norethindrone and ethinyl estradiol, daily bleeding diaries, follicle growth, changes in cervical mucus and progesterone levels drawn at 7- to 10-day intervals. RESULTS: Treatment with St. John's Wort was associated with a significant 13-15% reduction in the dose exposure from the contraceptive. Breakthrough bleeding increased in the treatment cycles, as did evidence of follicle growth and probable ovulation. CONCLUSION: St. John's Wort is associated with increased metabolism of norethindrone and ethinyl estradiol, breakthrough bleeding, follicle growth and ovulation. Women using OCs should be cautioned that St. John's Wort might interfere with contraceptive effectiveness.

Adult↗

[Cell culture for active Chlamydia trachomatis infection in a population of symptomatic women in Abidjan].

The object of our study has been to assess Chlamydia trachomatis prevalence among symptomatic women in Abidjan and to identify issues related to the use of cell culture methods in a tropical laboratory. 1522 women with vaginal discharge were enrolled in a cross sectional study. One endocervical swab was taken per woman and inoculated into cycloheximide treated Mac Coy cells. Elementary bodies were detected by direct fluorescent antibody (DFA). The isolate rate of Chlamydia trachomatis by cell culture was estimated to 86%. The prevalence of chlamydial infection among symptomatic women was 10.8%. Culture was influenced by presence of blood or cervical mucus in the sample. 206 samples gave no results because of blood or cervical mucus. During this study repeated contaminations of cells with facultative bacteria were noted and disposing of a sufficient number of cells was not easy.

Blood↗

Changes in enzyme levels in human cervical mucus during the menstrual cycle.

Cervical mucus is a complex secretion produced by the endocervical epithelium. A comparative analysis of 30 enzymes was carried out on 15 women during their periovulatory period (days 10 to 16 of the menstrual cycle). Nine different enzymes were always found to be present in the cervical mucus. Changes in enzymatic activity were further studied in two of the women, one with a normal (27-day) and the other with a short (20-day) menstrual cycle. The data were compared with those for the same enzymes in the saliva of these two women. Each woman's ovulation time was determined through hormone measurements in urine and blood, as well as by recording basal body temperature and by Billing's method. Our results indicate that a maximum peak occurs for several enzymes during the periovulatory period.

Adult↗

Contraceptive action of continuous low doses of norgestrel.

Norgestrel in a dose of 50 mug. was administered daily to 188 women during 2,250 menstrual cycles. Only two pregnancies occurred because of failure of the method, giving a failure rate of 1.1 pregnancies per 100 woman-years. The method was acceptable to most of the subjects, and side-effects, other than menstrual irregularity, were minimal; 68% of the cycles were 28 +/-5 days in length and 21% were less than 23 days. There did not appear to be any increased incidence of amenorrhoea. The antifertility action of continuous administration of low doses of progestogen may be due to an effect on corpus luteum function, in addition to the effects on cervical mucus, endometrium, and tubal transport of ova. The optimum dose of norgestrel appears to be in the range of 50-75 mug./day.

Adolescent↗