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The aim of our preliminary study was to compare the levels of total local sIgA and IgG with activity of detected sperm antibodies in ovulatory cervical mucus (OCM).Department of Gyneacology and Obstetrics, Medical School and Faculty Hospital, Charles University, Plzen.We screened samples of OCM from 12 pacients aged 26-31 (29,6 years on average) by radial immunodifusion (RID) to determine s IgA and IgG. Indirect MAR test was used for detection of spermagglutinationg antibodies.We found out by RID the average concentration of sIgA in OCM 567,84 mg/l (0 -1250,47) and the average concentration of IgG in OCM 23,57 mg/l (8,74-47,99). Antibody activity against sperm cells dominates in IgA with 6 pacients, in IgA with 1 patient, in IgA and IgG together with 1 infertile woman and in IgA and IgM isotypes together with 1 patient. No local sperm antibodies were determined with 3 patients.We proved the hypothesis, that the levels of spermagglutinating antibodies do not correlate with findings of total sIgA and IgG in OCM with our patients.

The aim of this study was to compare levels of PGE2 in cervical ovulatory mucus in patients with various frequency of spontaneous miscarriages, and in healthy fertile women.Special Consultation for Immunology of Reproduction and Research Laboratories for Reproductive Immunology, Department of Gynaecology and Obstetrics, Medical Faculty of Charles University and Faculty Hospital, Pilsen.We used commercial, high sensitive method of ELISA (PGE2 HS, RD Systems) for detection of PGE2 in cervical ovulatory mucus. Statistic analysis (T-test, ANOVA-test and Kruskal-Wallis test) also provided our study.It was taken in total 120 measurements. The RD Systems company does not mention physiological levels of PGE2 in cervical ovulatory mucus. Therefore we compared all results of our studied groups with average concentration (197356 pg/ml PGE2) gained from control group. We found that in patients with one miscarriage PGE2 levels are approximately 6 times higher, with two miscarriages 13 times higher, and in patients with three and more miscarriages even 21 times higher than controls (p0.0327, alpha 5%).In present literature does not exist any study concerning of PGE2 in cervical ovulatory mucus. We prove that some patients mainly with repeated "unexplained" miscarriages have ovulatory levels of PGE2 significantly higher than controls.

Study of antibodies against laminin-1 in IgG in patients with habitual abortion.Prospective study.Commercial ELISA method for detection of IgG antibodies against laminin-1 in ovulatory cervical mucus, in sera in women with repeated spontaneous abortions, in sera in patients during spontaneous miscarriages.Positive levels of antibodies against laminine-1 in IgG in sera were found in patients during spontaneous abortions, as to ovulatory mucus of infertile women in time out of pregnancy, we did not find any positivity, only in one serum sample in patient with three pregnancy losses in her history.Antibodies against adhesive molecule as laminin-1 presents, can participate during diagnosis of repated spontaneous miscarriages in context of neuro-endocrinne-immune dysregulation, which is not seen during physiological pregnancy.

To study serum antibodies against annexin V and against other phospholipids in women with fertility failure.Prospective study.Department of Gynecology and Obstetrics, Institute of Imunology and Alergology, Institute of Forensic Medicine, Charles University and Faculty Hospital, Pilsen, Czech Republic.We investigated sera from a total of 1550 women aged 26-41 years who attended the Division for Infertility and Reproductive imunology, in the Department of Obstetrics and Gynecology, Charles University, and Faculty Hospital, Pilsen, Czech Republic. ELISA was used for detection of antibodies against annexin V, phosphatidic (ph)-acid, ph-ethanolamine, D-L glycerol, ph-inositol, L-serine, cardiolipine, beta-glycoprotein, and prothrombine), antibodies against zona pellucida by passive hemmagglutination. Antibodies against annexin V in ovulatory cervical mucus were studied only in 47, and in follicular fluid in 168 women. Eighty-four fertile healthy women were included in the control group.Antibodies against annexin V were found in 13% (in 170 infertile women). Anamnestic data showed two or more unexplained pregnancy losses in 48 (28.2%), 108 (63.5%) patients had two or more unsuccessfull in vitro fertilization, polycystic ovaries (PCO) was diagnosed in 3 (1.7%), and endometriosis in 11 (6.6%) of them. The increased levels of antibodies against ph-inositole (28.8%), and L-serine (19.4%) were detected. 353% of patients had positive levels against zona pellucida. In ovulatory cervical mucus only in six (6/47) antibodies against annexin V were detected, in 5.55% in follicular fluid (6/108).Our study also demonstrated serum positivity against annexin V as a significant risk factor mainly in patiens with repeated reproductive failure.

To evaluate the presence and activity of local sperm antibodies and electrophoretic analysis of ovulatory cervical mucus (OCM) to prove the correlation of results of Kremer mucus sperm capillary penetration test and i-MAR (mixed antiimmunoglobulin reaction) test with SDS-PAGE.Retrospective study.Department of Obstetrics and Gynecology, Medical Faculty of Charles University, Pilsen.94 patients aged 22-40 (average age 32.3 years) were chosen for our study. Ovulatory cervical mucus was taken from uterine cervical canal in Consultation for Reproductive Immunology. Kremer test and indirect mixed antiglobulin reaction test for IgG, IgA, IgM and IgE were used for detection of sperm antibodies. For the SDS-PAGE analysis, OCM was incubated with sodium dodecyl sulfate. We studied separated protein fractions from OCM.Sperm-capillary ovulatory mucus penetration test (Kremer) was 0-10 cm/hour in our group (average value in patiens without sperm antibodies was 2.43 cm/hour, with sperm agglutinating antibodies 1.4 cm/hour), significant levels (45%) of spermagglutinating antibodies were detected in IgA in 6 patients (6.38%), IgG in 5 (5.32%) patients, sperm-cytotoxic levels (IgA and/or IgG) in 5 patients (5.32%); levels of IgE less than 30% in 3 patients (3.19%). Individual immunological factors gained by SDS-PAGE showed the spectrum of various molecular weights with range of 14.4- 350 kDa. The presence of IgG and/or IgA was in 53 cases (56.38%), with 13 ASA positivities (24.53% correlation with i-MAR test) and no ASA activity in 40 cases (75.47%); 6 ASA positive patiens (31.58%) were not detected by SDS-PAGE.Analysis of 94 OCM by SDS-PAGE showed several significant correlates, but their specifications will be based on further immunoblot research.

Detection of antizonal antibodies (aZPs) in cervical ovulatory mucus and in serum in patients with fertility failure.Retrospective study.Department of Gynaecology and Obstetrics, Medical Faculty of Charles University, and Faculty Hospital, Plzen.Detection of aZPs (passive hemagglutination test) in cervical ovulatory mucus (OM) and in serum from 105 patients with reproductive failure and in 25 fertile healthy women (control group).Positive levels of aZP in OM were found in 79 (75.2%) patients, in 56 of them aZP were observed in OM and in serum together, only ovulatory aZPs in 23 (21.9%), and only serum aZPs in 18 (17.1%) were found. aZPs predominate in 47 patients (of 48 women) with more than two in vitro fertilization processes, in thirty of them aZPs in serum and in OM together were observed. On the other hand, repeated spontaneous miscarriage and obesity have no connection with detection of aZPs in our patients. CONCLUSION. The increased levels of serum and OM aZPs are very often seen in patients after repeated unsuccessful IVF. This very important result in reproductive immunology means to go on the micromanipulation technics combined with immunosuppression in the next planned IVF procedure.

The penetration of native and cryopreserved human spermatozoa into cervical mucus--the Kremer's test.Retrospective study.Department of Obstetrics and Gynaecology, Medical Faculty, Charles University and Faculty Hospital, Plzen.Human cervical mucus was collected from 73 women visiting the Division of immunology of reproduction, Department of Obstetrics and Gynaecology in Plzen. Native spermatozoa was obtained from the patients of the Division of Immunology of Reproduction as well. Cryopreserved only in its seminal plasma was obtained from the spermabank of our department. The distance of penetration in centimetres from the start was examined in inverse microscopy after 30, 60, 90 and 90 minutes. Also the character and duration of the sperm motility was analyzed.The penetration of native spermatozoa was higher than the penetration of cryopreserved spermatozoa in each case. The native spermatozoa had a higher penetrability, motility and life-ability. Spermatozoa preserved only in its seminal plasma had the parameters demonstrably lower. Nevertheless these spermatozoa can be successfully used for homologue or heterologue insemination or for IVF because these spermatozoa do not loose its enzymatic and remaining energetic equipment by penetration the cervical canal.The Kremer's test belongs to the reliable methods of penetration ability of native and cryopreserved sperms.

Applying the method of absorption atom spectrometry (AAS), the contamination of cervical mucus by chemical elements (Cd, Pb, Hg, Cr, Cu, Zn) was demonstrated in the oestrus period of cows in different ecological agricultural regions of the North Moravian region. The results of observation revealed only a statistically significant difference (P less than 0.05) in Cd contents in the test groups; test group mean = 0.015 micrograms.g-1 Cd, control group mean = 0.006 micrograms.g-1. Cr findings in cervical mucus are of priority importance, the same applies to Cu findings. Zinc was found to be an element influencing negatively the conception of cows. The Zn values in cervical mucus of nonpregnant cows were demonstrated to be significantly higher (P less than 0.01) (conception--group = 0.841 micrograms.g-1, conception + group = 0.219 micrograms.g-1 Zn. So called sum of chemical elements: Cd + Pb + Hg + Cr + Cu + Zn was proposed and evaluated as a picture of the total contamination of cervical mucus. This characteristic was also influenced by the Zn findings in the group of pregnant and nonpregnant cows. Zn: Cd antagonism in cervical secretions of barren and pregnant cows was statistically highly significant (P less than 0.01). Chemical elements in cervical mucus were determined for the first time in a digested sample of cervical secretion. The correlation coefficients used for the cows (n = 99) from a contaminated region showed that the content of chemical elements in cervical secretion was not influenced by the age of cows. The content of chemical elements in cervical secretion was not increasing with the age.

Cervical secretion was collected from 99 dairy cows of the Black Pied Lowland breed at an age of 24 to 109 months with a minimum performance of 4000 kg of milk annually in spontaneous oestrus 42 days post partum at the minimum. It was demonstrated by the radioimmunological determination of progesterone in skim milk that from all the breeding cows (n = 99) the oestral secretion was collected during an optimum insemination time (x = 0.19 ng.ml-1 progesterone). Of the statistical set of 99 cows, 55 (i.e. 56.57%) got in calf and 43 (i.e. 43.43%) remained barren. As found, the conception results were not influenced by the rank of the insemination after parturition and the conception capacity of the cows was not influenced by the length of time from parturition to the respective inseminations. It was demonstrated in our investigation that the conception capacity of the cows increases as the time of sperm survival prolongs in the cervical secretion in vitro. The highest conception capacity (80%) was recorded in the cows whose cervical secretion was free of spermiotoxic action. In such cases the motility of the spermatozoa in the oestral mucus was maintained at a temperature of 38 degrees C for longer than 300 minutes. On the other hand, it was demonstrated that the conception capacity of the cows declines with shortening time of sperm survival in the cervical mucus.

31 women were administered a modified mintablet contraceptive, consisting of .3 mg of norethisterone for an average of 10.6 months. During this period changes in the cervical mucus were analyzed. Changes in the protein fractions were revealed, which were characterized by a widening of the spectrum of albumins in the area of globulins and some special fractions. These changes, elicited by the contraceptive effect of the gestagen preparation, were confirmed in comparison to a control group through a permeation test.

The pattern of changes in the trypsin-inhibition activities (TIA) of blood plasma, cervical mucus and in the morphological structure of cervix was studied in ewes exposed to 60Co radiation for seven and five days, the radiation doses being 6.7 Gy and 4.8 Gy (700 and 500 R). During exposure, the group of ewes irradiated with 4.8 Gy was given the Roboran vitamin premix in addition to standard feed. After termination of irradiation the animals in this trial were given the ampicillin antibiotic (5250 mg). TIA was determined from the retardation of the hydrolysis of the synthetic substrate N-alpha-tosyl-p-nitroanilide (TAPA) by bovine trypsin; the TIA was expressed as the percentage of inhibited trypsin. As found on the day of the termination of irradiation, almost all the studied TIA values of blood plasma and cervical mucus were increased in the irradiated animals, the range being from 103.1 to 155.0% of the levels for non-irradiated ewes. A reduction was recorded only in total TIA of blood plasma in the group irradiated with the dose of 6.7 Gy (83.1% of the values for non-irradiated animals). In the group of animals irradiated with 4.8 Gy and fed no Roboran, the TIA of cervical mucus was observed to decrease to 92.4%. When evaluating the average TIA of different test groups throughout the experiment it was found that all the studied TIA values of the irradiated animals increased within a range from 104.1 to 184.0% of the values for untreated ewes. It was only in the average values of total TIA in blood plasma that, after irradiation with 6.7 Gy, a decrease to 92.7% of the level without irradiation was recorded. It was found during the study of changes in the proportions of glands in the stroma and changes in epithelium thickness in the mucous membrane of cervix uteri that the irradiated ewes had the epithelium thickness reduced to 95.3%-65.5% and that their stromal gland number decreased to 75.4%-79.7% of that recorded in non-irradiated animals. It was only in the group given a Roboran supplement that an increase to 123.7% of the gland number for untreated ewes was recorded on the tenth day after the termination of irradiation.

59 users of the polyethylene (IUD) intrauterine device made in Czechoslovakia were investigated for the presence of urea in the cervical mucus, and for 29 of them, in the intrauterine fluid as well. Urea in the cervical slime was detected in 43 women and in 42 of them also in the uterine cavity fluid. The results were processed graphically and statistically. It was found that the level of urea was to some extent dependent on the menstrual cycle, with maximum values just prior to menstruation. There was, however, no difference in the level of urea between the IUD users and the control group, the levels roughly corresponding to the physiological values of urea in the serum. It is thus unlikely that the cytolytic effect of increased levels of urea due to the breakdown of proteins in the endometrial tissues is the result of the contraceptive effect of IUDs. (Authors' modified)