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[Isolation and purification of an antibacterial polypeptide from human cervical mucus].

OBJECTIVE: This study was made to isolate and purify antibacterial polypeptides from human cervical mucus. METHODS: Human cervical mucus was collected from human healthy subjects, and the acid-soluble extract was obtained by solving the mucus with 5% acetic acid in the presence of protease inhibitor. By using gel overlay technique, one protein band that was potently antibacterial against E. coli ML-35P was identified. To purify the antibacterial polypeptide, the preparative acid-urea gel electrophoresis and Reverse Phase HPLC were performed. RESULTS: A purified antibacterial polypeptide was obtained and named HCP-1. Tricine-SDS-PAGE indicated that the relative molecular mass of HCP-1 was around 14 x 10(3). Agar radial diffusion assay showed that the purified polypeptide had antibacterial activities against E. coli ML-35P and Pseudomanas aeruginosa ATCC65922. CONCLUSION: HCP-1 may be a new effector molecule of human cervical mucus.

Adult↗

[Study of antmicrobial mechanisms of human cervical mucus: isolation and characterization of antibacterial polypeptides].

OBJECTIVE: To isolate and purify antibacterial polypeptides from human cervical mucus. METHODS: Human cervical mucus was collected from human healthy subjects and acid-soluble extract was obtained by solving the mucus with 5% acetic acid in the presence of protease inhibitors. The antibacterial components were identified by Agar radial diffusion assay and gel overlay technique. For further purification, Preparative acid-urea gel electrophoresis and Reverse Phase HPLC were performed. The N-terminal sequencing and degenerate PCR-directered cDNA cloning were performed. The E. coli-based recombinant product was prepared and its antibacterial property was determined by minimal inhibitory concentration and minimal bactericidal concentration. RESULTS: A purified antibacterial polypeptide was obtained. Agar radial diffusion assay showed that the purified polypeptide had antibacterial activities against E. coli ML-35P and Pseudomanas aeruginosa ATCC 27853. The N-terminal amino acid sequence and its full length of cDNA were identical to High Mobility Group Chromosal protein N2 (HMGN2). The purified recombinant HMGN2 was obtained. Its MIC against E. coli ML-35p and P. aeruginosa ATCC27853 were 10.42 microg/ml +/- 3.13 microg/ml and 27.78 microg/ml +/- 8.33 microg/ml respectively, which were equal to human neutrophil defensins HNP, and the MBC were 20.83 microg/ml +/- 6.25 microg/ml and 55.56 microg/ml +/- 16.67 microg/ml respectively. CONCLUSION: HMGN2 may be another antibacterial effecter in the defense mechanisms of human cervical mucus.

Adult↗

Early pregnancy factor in cervical mucus of pregnant women.

PROBLEM: This study was carried out to determine the presence of early pregnancy factor (EPF) activity in cervical mucus of pregnant women and explore the relationship between EPF activity in sera suggested it and in cervical mucus. METHOD OF STUDY: Cervical mucus and sera were collected from 60 pregnant women in different terms and 25 non-pregnant women, respectively. EPF activity was detected by rosette inhibition assay. RESULTS: EPF activity did exist in cervical mucus of pregnant women and was similar both in cervical mucus and sera. CONCLUSIONS: We report here for the first time the EPF activity in cervical mucus of pregnant women. The parallel change observed in sera suggested it might be another useful index in evaluating embryo viability.

Cervix Mucus↗

Health workers' guide to natural family planning.

Health workers can help couples avoid unwanted pregnancies and choose when to have a baby by teaching them how to use natural family planning (NFP). A new WHO document for health workers describes how NFP works,. how effective it is and how couples can be taught to use it. The guide deals with 4 natural methods of family planning, all of which relate to identifying just when a woman is fertile or infertile during her menstrual cycle. The rhythm method requires the woman to calculate her fertile days according to the length of her menstrual cycles; the cervical mucus method makes the same calculation according to the amount and color of mucus in the vagina; the basal body temperature method relies on recording temperature changes during the menstrual cycle; and the symptothermal method bases the calculation on cervical mucus, body temperature and other signs of fertility such as tenderness of the breasts or abdominal pain. WHO studies of NFP show that the cervical mucus method works in 97 case out of 100 where couples use the method correctly. WHO believes that NFP is suited to couples who do not wish to use other contraceptive methods for family reasons or because of religious beliefs.

Delivery of Health Care↗

Immunoglobulins, proteinase inhibitors, albumin, and lysozyme in human cervical mucus. I. Communication: hormonal profiles and cervical mucus changes--methods and results.

The serum levels of luteinizing hormone (LH), estradiol-17beta, and progesterone were determined simultaneously with the concentrations of immunoglobulin (Ig) G, IgA, C'3, alpha1-antitrypsin, inter-alpha-trypsin inhibitor, alpha1x-antichymotrypsin, albumin, and lysozyme in cervical mucus during nine ovulatory cycles. Spinnbarkeit and ferning were also assessed, and the basal body temperature was measured and recorded during these cycles. The profiles were synchronized according to the LH peak. The midcycle period, characterized by the rapid increase and decline of estrogen and the beginning rise of progesterone, shows a prounced minimum of immunoglobulins, C'3, proteinase inhibitors, albumin, and lysozyme in cervical mucus, which is known to be most receptive to sperm penetration at this time. Although the variation of cervical mucus values is considerable during the early proliferative and the luteal phases, the midcycle values appear to be constantly low, showing slight differences among the profiles of the different parameters. The statistical evaluation and the assessment of the significance of parameters for ovulation detection and the assessment of the fertile period as well as the correlation of these parameters with basal body temperature will be the subject of the second communication of this series.

Adult↗

Fertility awareness / natural family planning for adolescents and their families: report of multisite pilot project.

Fertility awareness is experiential learning about cyclic fertility. This awareness, used as a family planning method, differs from contraception because it does not isolate the procreative capacity of either partner. The acceptability and effect of teaching fertility awareness on teen sexual activity and decision making was tested in a multisite pilot program which taught fertility awareness via the prospective marker of the cervical mucus (ovulation method of natural family planning). 200 US and 35 Guatemalan volunteer women ages 15-17 in a structured 1 year curriculum, monitored cycle charting and explored the implications of experiencing one's signs of fertility. Control subjects were recruited from the general population and from family planning clinics. 9% of the US study group were sexually active prior to entry. By cycle 12, 1/2 had discontinued activity. Conception rate was 0.0044. The continuation rate dropped from 90% at cycle 7 to 71% at cycle 8 due to scheduling constraints for 2 classes and to 57% at cycle 12. Postprogram follow-up of early leavers showed only 1/3 the expected rate of onset of sexual activity and pregnancy. Parent involvement correlated positively with postponement and/or discontinuation of sexual activity. Reported movement away from peer group pressure appeared 3 months after entry.

Adolescent↗

[Use-effectiveness of natural family planning by ovulation method].

Cervical mucus (CM) peak and plasma LH surge were examined in 19 women. The consistency of the CM peak and ovulation after the plasma LH surge demonstrated that as an indicator of natural family planning (NFP), the CM can scientifically predict the occurrence of ovulation. Based on these results, 300 healthy women were recruited into this study by self examination of CM. A total of 3,393 cycles was observed. There were 29 pregnancies reported, and the continuation rate was 90.3/100 women year. It showed that the NFP is an acceptable, safe and effective birth control method for Chinese women.

Adult↗

Biochemical changes in the cervical mucus after application of laminaria tent.

BACKGROUND: This research was aimed to study the biochemical changes in cervical mucus after application of laminaria tent. MATERIAL AND METHODS: Cervical mucus was collected before and after insertion of laminaria tent from 20 normal pregnant women in the first and second trimesters. Interleukin-1 beta, IL-8, collagenase activity, elastase activity, PGE2 and PGF2 alpha were measured in the collected samples. Laminaria tent was also inserted in one vaginal cervix of five non-pregnant rabbits and the clinical and histological changes were compared with five non-treated rabbits. Collagen contents were assessed histologically and Leukocyte number was counted. RESULTS: IL-1 beta and IL-8 and elastase activity were significantly increased after insertion of laminaria (p<0.0001 and 0.001 respectively). Collagenase activity was not significantly changed. Whereas concentration of PGE2 and PGF2 alpha in the mucus were markedly increased (p<0.0001 and 0.001 respectively). Collagen contents were significantly decreased in laminaria and opposite side cervices (p<0.0001). No significant changes in leukocyte count could be observed in treated and non treated cervices. CONCLUSIONS: Insertion of laminaria leads to increase concentration of IL-1 beta, IL-8, PGE2, PGF2 alpha and elastase activity. Mechanical stretching of the cervix and biochemical changes in cervical mucus may explain the mechanism of cervical ripening induced by laminaria.

Animals↗

IL-1beta, IL-6, TNFalpha, fetal fibronectin, and endotoxin in the lower genital tract of pregnant women with bacterial vaginosis.

BACKGROUND: In our studies on women with bacterial vaginosis (BV) in early pregnancy a strong association has been found between BV and the levels of endotoxin or interleukin-1alpha (IL-1alpha) in the lower genital tract. In the present study we investigated if an association could be found between BV and other cytokines (IL-1beta, IL-6, tumor necrosis factor alpha, TNF) or fetal fibronectin (FFN). The cytokine-inducing capacity of endotoxins present in the cervical mucus was explored in a monocytic cell assay. METHODS: Cervical mucus or cervicovaginal fluid was collected from women with (BV) and without BV (nonBV) attending a family planning unit for first trimester abortion. The concentrations of IL-1beta, IL-6, TNF and FFN were determined by quantitative enzyme immunoassays. TNF was determined in 63 women (BV, n=25) out of whom 37 (BV, n=11) were analyzed for IL-1beta and the remaining 26 for IL-6 (BV, n=14). FFN was determined in another 36 women (BV, n= 19). The cytokine-inducing capacity of endotoxin-containing cervical mucus and purified endotoxin of Prevotella bivia were studied by an in vitro cell assay using a human monocytic cell line (THP-1). RESULTS: IL-lbeta and IL-6 were found in almost all women. The levels of IL-1beta, but not IL-6, TNF or FFN, were significantly increased in women with BV compared with the nonBV women (p<0.05). Purified endotoxin from P. bivia, and cervical mucus from BV women containing high levels of endotoxin were able to induce a cytokine response (IL-6) in monocytic cells in vitro. CONCLUSION: BV is associated with increased levels of IL-1beta in the lower genital tract of pregnant women in the first trimester. The ability of BV-associated endotoxins to induce cytokine production in monocytic cells may partly explain the increased IL-1beta levels.

Adult↗

[Charactierization of a new antibacterial polypeptide isolated from human cervical mucas].

OBJECTIVE: To identify a new antibactrial polypeptide HCP-1 isolated from human cervical mucus. METHODS: HCP-1 was isolated and purified from human cervical mucus, and N-terminal sequence of HCP-1 was determined. A degenerate primer was designed according to CodeHop methods, and an "anchor-oliga-dT primer" was used for the synthesis of cDNA. Using the degenerate primer and anchor-primer, cDNAs were amplified by PCR. The PCR products were cloned, sequenced, and analyzed by biological software. RESULTS: N-terminat sequence of HCP-1 was PKRKAEGDAK. The full length of HCP-1 cDNA was isolated and of which the sequence was the same as HMG-17. OMIGA software analysis indicated that this molecule contained an alpha-helix region. CONCLUSION: The new antibacterial polypeptide isolated from human cervical mucus is HMG-17. It may play a role in the human cervical mucus and the alpha-helix domain may be its antibacterial activity region.

Adult↗

A prospective multicentre trial of the ovulation method of natural family planning. III. Characteristics of the menstrual cycle and of the fertile phase.

Seven hundred twenty-five women of proven fertility recorded the presence of cervical mucus at the vulva in 7514 menstrual cycles. The mean cycle length of the 6472 "normal" cycles was 28.5 days (standard deviation +/- 3.18). The peak day of mucus discharge was the last day of slippery, raw-egg-white-like mucus and occurred on average on day 15 (+/- 2.6). The fertile period was defined as any day on which mucus was reported before the peak day until 3 days after the peak. Its mean length was 9.6 (+/- 2.6) days. The probability of pregnancy was maximal on the peak day and declined on the days before and after the peak.

Adult↗

Identification of tobacco-specific carcinogen in the cervical mucus of smokers and nonsmokers.

BACKGROUND: In 1996, an estimated 15,700 new cases of cancer of the uterine cervix and 4,900 deaths from this disease were expected to occur in the United States. In a recent international study, human papillomavirus DNA was found in more than 90% of cervical tumor specimens examined, irrespective of the nationality of the patients from whom the samples were obtained. Although infection with human papillomavirus is the major known risk factor for the development of cervical cancer, it alone is not sufficient. Other etiologic factors that have been associated with this disease include deficiencies in micronutrients, lower socioeconomic status, oral contraceptive use, and cigarette smoking. Several compounds from cigarette smoke (nicotine and its major metabolite, cotinine) have been identified in cervical mucus, and the occurrence of smoking-related DNA damage in the cervical epithelium has been documented. PURPOSE: This investigation was conducted to determine for the first time whether carcinogenic tobacco-specific N-nitrosamines are present in the cervical mucus of cigarette smokers and of nonsmokers (most likely as a result of environmental exposure). METHODS: Cervical mucus specimens from 15 smokers and 10 nonsmokers were subjected to supercritical fluid extraction with the use of carbon dioxide that contained 10% methanol, and the resultant extracts were analyzed for tobacco-specific nitrosamines by use of a very sensitive method that involved gas chromatography and mass spectroscopy analyses. RESULTS: In a total of 16 samples obtained from 15 women who were current smokers (two samples from the same woman), we detected the tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) at concentrations that ranged from 11.9 to 115.0 ng/g of mucus. Only one of a total of 10 cervical mucus specimens obtained from 10 women who claimed to be nonsmokers did not contain detectable NNK, and NNK concentrations ranged from 4.1 to 30.8 ng/g of mucus in the specimens from the remaining nine women. The concentrations of NNK in specimens from cigarette smokers were significantly higher than those from nonsmokers (mean +/- standard deviation: 46.9 +/- 32.5 ng/g of mucus versus 13.0 +/- 9.3 ng/g of mucus; two-tailed Student's t test, P = .004). CONCLUSION: The cervical mucus of cigarette smokers contains measurable amounts of the potent carcinogen NNK. This compound represents the first tobacco-specific carcinogen identified in this physiologic fluid of women who smoke cigarettes. The presence of NNK in the cervical mucus of nonsmokers is likely due to environmental exposure or to the fact that some of the subjects in this study may not have revealed that they occasionally smoked cigarettes. IMPLICATIONS: The presence of NNK in human cervical mucus further strengthens the association between cervical cancer and tobacco smoking.

Adult↗

Effect of two anti-estrogens, clomiphene citrate and tamoxifen, on cervical mucus and sperm-cervical mucus interaction.

OBJECTIVE: To compare the effect of two ovulation-inducing agents, clomiphene citrate and tamoxifen, on cervical mucus and sperm-cervical mucus interaction. SUBJECTS AND SETTING: Forty couples with unexplained infertility attending infertility clinic. METHODS: Cervical mucus scoring and postcoital test done using the Moghissi system in a spontaneous cycle (control cycle) and with clomiphene citrate or tamoxifen (study cycles). RESULTS: Clomiphene citrate significantly decreased cervical mucus production, whereas tamoxifen significantly improved the total score. CONCLUSION: Tamoxifen is a better drug than clomiphene for ovulation induction in women with poor cervical mucus quality.

Adult↗

Relationship between cervical mucus interleukin-8 concentrations and vaginal bacteria in pregnancy.

PROBLEM: High interleukin (IL)-8 concentration in cervical mucus in the second trimester is a risk factor for premature birth. We investigated the relationship between vaginal pathogens and IL-8 in cervical mucus. METHOD OF STUDY: In 501 women with single pregnancy, vaginal secretions were cultured for bacteria and cervical mucus IL-8 concentrations were measured between 20 and 24 gestational weeks. RESULTS: Lactobacillus species were detected in 56.0% of 84 subjects with high IL-8 (> or =377 ng/mL), significantly less often than in 417 subjects with IL-8 below 377 ng/mL (84.7%; P < 0.0001). Anaerobic pathogens were detected in 83.3% of high IL-8 subjects, significantly more often than in normal IL-8 subjects (43.9%; P < 0.0001). By multivariate analysis, cervical IL-8 was significantly high only in subjects without Lactobacillus species; they showed a significantly higher prematurity rate than Lactobacillus-positive subjects. CONCLUSIONS: Absence of vaginal Lactobacilli was associated with increased cervical IL-8 and increased risk of premature delivery.

Adult↗