Postcoital Sperm Assessment Comparative Study.

This postcoital sperm assessment study was performed over a 10 month time period (November 2014-August 2015). Fifteen couples enrolled in the study. The study was a non-blinded, non-randomized, single-center comparison study comparing The Stork® OTC (Rinovum Women's Health, Monroeville, PA) to natural intercourse (NI), using the subjects as their own control/baseline. This was an efficacy study designed to compare the number of sperm in the cervical mucus following the use of The Stork OTC conception aid with the number of sperm in the cervical mucus following natural intercourse. Subjects used both The Stork OTC conception system and the natural intercourse method to evaluate concentrations of sperm in the cervical mucus. Post-coital test (PCT) data was collected demonstrating higher concentrations of sperm within the cervical mucus with The Stork OTC conception system versus natural intercourse for 85% of test subjects in this study. Of the 15 couples enrolled in the study, 2 were lost to follow-up. Mean age for male subjects was 31.7 +/ 5.4 years of age and mean age for female subjects was 29.7+/- 5.4. The average sperm score value of the 85% of test subjects with higher sperm concentrations from The Stork OTC was 3.23 times the score value of sperm concentration compared to natural intercourse. The remaining 15% of test subjects showed no change in sperm score value between The Stork OTC and natural intercourse.

[Prediction of complications of I trimester of pregnancy in women with distressed reproductive history by detection of transcervical trophoblastic cells].

The present study investigated the possibility of using trophoblastic (fetal) cells in cervical mucus of pregnant women with complicated reproductive history to predict the course of early I trimester of pregnancy. We examined 86 women at 5-6 weeks of gestation, 56 of them with normal pregnancy, 12 with missed abortion, 18 with tubal pregnancy. As a control we used 21 non-pregnant womеn with primary infertility. Trophoblastic cells we detected by immunohistochemical method and then counted. It was revealed that in the normal course of early I trimester transcervical trophoblastic cells were detected in 4 times more likely than in missed abortion and 5 times more often than in women with tubal pregnancy. In women of the control group, who never had pregnancies, trophoblastic cells were not detected. Thus, the determination of the number of transcervical trophoblastic cells can be used in the complex diagnosis of pathological course of I trimester of pregnancy in women with complicated reproductive history.

Effect of intrauterine infusion with liquid paraffin on phagocytes migrating to mucus of external os of the cervix in cows.

The objective of this study was to investigate the effect of intrauterine infusion with liquid paraffin (LP) on phagocytic migration into the uterus of cows. Smears of swab samples of the external os of the cervix and discharges collected inside the vagina were obtained in multiparous dairy cows (n = 10) that had been infused with 50 ml of LP (LP group: n = 5) or physiological saline (PS group: n = 5) on day 10 or 11 after ovulation (day 0: ovulation). The samples were collected for cytological examination 0 (just before), 0.25, 1, 2, 3, 4, 6, 8, 12 and 24 h after LP or PS infusion and then at daily intervals until subsequent ovulation. The number of neutrophils increased significantly (p < 0.05) for 8 days compared with the pre-infusion level in the LP group and for 2 days in the PS group. The average numbers of neutrophils in the LP group were significantly (p < 0.05) greater than those in the PS group on 3, 4, 5 and 8 days after infusion. The number of monocytes from 6 h to 8 days after LP infusion was significantly higher than that before infusion (p < 0.05). The average numbers of monocytes at 4 and 6 h and 1 day after infusion in the LP group were significantly higher than those in the PS group. These findings indicate that LP stimulates phagocytic migration into the uterine lumen in cows and that LP infusion into the uterus might enhance uterine defence mechanisms during uterine infection.

Impact of estrous synchronization methods on cellular proportions in cervical mucus and serum hormone concentrations.

The objective of this study was to examine cytological changes of cervical mucus following the induction of estrus with intra-vaginal drug release (CIDR) devices in dairy cows. Sixty healthy Holstein Frisian cows, averaging 80 (+/-10) days post-partum, were selected from a commercial dairy farm around Shiraz. Cows in the control group were synchronized by the Ovsynch protocol. Cows in the second group (OV+CIDR) were subjected to the same Ovsynch protocol but in addition were administered a progesterone-releasing CIDR. Cows in the third group (OV+S-CIDR) were subjected to Ovsynch procedures but received the skeleton of a CIDR device, which did not release progesterone. Cows in the fourth group (E2+CIDR) received a progesterone releasing CIDR but were injected with estradiol benzoate. Cows in group 5 (E2+S-CIDR) received a CIDR skeleton and estrodial benzoate. CIDR devices were removed from cows in groups 2-4 and all cows were injected with PGF2alpha on day -3. Blood samples and cervical mucus discharges were collected from all cows on days -10, -3, 0 and 12 relative to AI. On the day of AI, the mean+/-S.D. percentage of neutrophils was significantly higher (p<0.05) in the S-CIDR+OV and S-CIDR+E2 groups than in Ovsynch group. Comparing the percentage of neutrophils in cervical mucus of Ovsynch group (less than 1%) with that of other treatment groups on the day of AI (from 5 to 39%) revealed the influential effect of a CIDR device on the reproductive tract. Results of the current study did not reveal hormonal effects but did identify mechanical effects of CIDRs on cell percentages in cervical mucus. The hormonal effects were probably masked by mechanical effects. Therefore, we were not able to confirm hormonal effects on proportions of different cells in cervical mucus. Consequently, additional research on hormonal effects and the mechanical effects of CIDR on the uterus is required.

[Cytological screening--the technique of cytological specimen taking and its influence on the quality of the method].

The aim of our research work was to evaluate if the way of taking material influence the quality of the Pap smear test. 2300 patients were examined by taking Pap smear test for 7 years period from 2000 till 2007. In 1000 patients endocervical brush /Cytobrush/ was used and after that Ayre spatula for taking ectocervical material. In 1300 patients Ayre spatula was used first and after that endocervical brush applied. The number of the Pap smears contaminated with blood, the Pap smears without endocervical component and the Pap smears with bad fixation were examined. There were more contaminated with blood pap smears in the group where endocervical brush was used first /156 patients vs 100, p = 0.004/. There was no significant difference in the number of bad fixated pap smears, the "thick" pap smears, the inflamated Pap smears in both groups. The quality of the technique for taking pap smears when Ayre spatula is used first and after that endocervical brush is better than the reverse way of obtaining cytological material from the cervix. With this technique we find more squamous intraepithelial lesions.

Corpora amylacea and molluscum contagiosum on a cervical pap smear.

When evaluating a Papanicolaou (Pap) smear, the cytology of the squamous epithelial cells is of utmost importance. This is what cytopathologists use to render a diagnosis, ranging from normal to atypical squamous cells of undetermined significance (ASCUS) to high grade squamous intraepithelial lesions, or even squamous cell carcinoma. However, occasionally the pathologist will run into microscopic noncellular material as in our case, such as corpora amylacea on the slides, or even uncommon viral inclusions such as Molluscum contagiosum (MCV). When these less common entities make their way on to a cervical Pap smear, it is important not only to correctly recognize them, but to understand their clinical implications as well. Diagn. Cytopathol. 2017;45:179-181. © 2016 Wiley Periodicals, Inc.

[Low-intensity laser effects on local immunity of female reproductive system affected with chlamidia infection].

Low-intensity laser impact on the main effector functions of the cervical secretion neutrophils in females suffering from Chlamidia infection has been studied. Dysfunction of neutrophil granulocytes of the cervical secretion was revealed. This manifested in the decreased number of phagocytes and lower rates of phagocytosis. The study of cervical secretion neutrophils has found that the content of active intracellular forms of oxygen was decreased in patients compared to healthy subjects. Low-intensity laser stimulated not only phagocytosis but also intracellular generation of active oxygen forms. Thus, low-intensity laser eliminates dysfunction of cervical secretion neutrophils.

Diffusion of Immunoglobulin G in Shed Vaginal Epithelial Cells and in Cell-Free Regions of Human Cervicovaginal Mucus.

Human cervicovaginal mucus (CVM) is a viscoelastic gel containing a complex mixture of mucins, shed epithelial cells, microbes and macromolecules, such as antibodies, that together serve as the first line of defense against invading pathogens. Here, to investigate the affinity between IgG and different mucus constituents, we used Fluorescence Recovery After Photobleaching (FRAP) to measure the diffusion of IgG in fresh, minimally modified CVM. We found that CVM exhibits substantial spatial variations that necessitate careful selection of the regions in which to perform FRAP. In portions of CVM devoid of cells, FRAP measurements using different IgG antibodies and labeling methods consistently demonstrate that both exogenous and endogenous IgG undergo rapid diffusion, almost as fast as in saline, in good agreement with the rapid diffusion of IgG in mid-cycle endocervical mucus that is largely devoid of cells. This rapid diffusion indicates the interactions between secreted mucins and IgG must be very weak and transient. IgG also accumulated in cellular debris and shed epithelial cells that had become permeable to IgG, which may allow shed epithelial cells to serve as reservoirs of secreted IgG. Interestingly, in contrast to cell-free regions of CVM, the diffusion of cell-associated IgG was markedly slowed, suggesting greater affinity between IgG and cellular constituents. Our findings contribute to an improved understanding of the role of IgG in mucosal protection against infectious diseases, and may also provide a framework for using FRAP to study molecular interactions in mucus and other complex biological environments.

Potential of syncytiotrophoblasts isolated from the cervical mucus for early non-invasive prenatal diagnosis: evidence of a vanishing twin.

BACKGROUND: Non-invasive methods to assess the foetal genome during pregnancy will provide new opportunities to offer pregnant women a more comprehensive genetic diagnosis of their established foetus. The aim of this study was to determine the presence and frequency of foetal cells in transcervical cell (TCC) mucus samples from pregnant women and determine their suitability for early prenatal diagnosis. METHODS: Syncytiotrophoblasts in aspirated TCC mucus samples were identified by immunostaining with the foetal-specific antibody NDOG1. Genetic analysis of foetal cells was performed by laser capture microdissection and quantitative fluorescent PCR (QF-PCR). RESULTS: In 116 of 207 (56%) TCC samples, abundant syncytiotrophoblasts were retrieved. However, when TCC samples were stratified for the presence of chorionic villous fragments, syncytiotrophoblasts were identified in 85 of 109 (78%) samples. Significant numbers of syncytiotrophoblasts were found in TCC samples collected between 6 and 9weeks of gestation (mean 741, range 25-2884). QF-PCR analysis of NDOG1 positive syncytiotrophoblasts and matching maternal DNA confirmed their foetal origin and correct foetal cell sexing was achieved in 97% of TCC samples. The one discordant sex diagnosis was associated with a dizygotic dichorionic twin pregnancy resulting from the implantation of a female T21 embryo and a normal male embryo, where the female T21 foetus had succumbed at 6weeks of gestation and was vanishing. CONCLUSIONS: Syncytiotrophoblasts can be successfully isolated from TCC samples and represent a suitable source of cells for genetic analysis of the established foetus in early pregnancy. The study highlights a vanishing twin as a potential cause for discordant non-invasive prenatal test results.

The Amount of MUC5B mucin in cervical mucus peaks at midcycle.

The physical character and amount of mucus secreted by the endocervix changes dramatically during the menstrual cycle to facilitate sperm migration at the time of midcycle ovulation. Mucins are highly glycosylated, high-molecular-weight proteins, which are the major structural components of the protective mucus gel covering all wet-surfaced epithelia, including that of the endocervix. We have previously demonstrated that the endocervical epithelium expresses messenger RNA (mRNA) of three of the large gel-forming mucins, designated MUC5AC, MUC5B, and MUC6, with mRNA of MUC5B predominating. Because mucin protein levels may be regulated posttranscriptionally, measurement of MUC5B protein levels with cycle are needed for correlation to mRNA levels. Measurement of specific mucin gene products within mucus secretions has been limited by availability of specific, well-characterized antibodies and by volume requirements of the isolation protocols for mucins, which include CsCl density centrifugation and fraction isolation. To measure MUC5B protein within the cervical mucus through the hormone cycle, we developed a polyclonal antibody specific to the mucin. The antibody, designated no. 799, is to a synthetic peptide mimicking a 19-amino-acid segment of an intercysteine-rich region within the D4 domain in the 3' region of the MUC5B protein. It recognizes native as well as denatured MUC5B on immunoblot, is preadsorbable with its peptide, and binds to apical secretory vesicles of epithelia expressing MUC5B. We used the MUC5B antibody along with a cervical mucin standard cervical mucin isolate in enzyme-linked immunosorbent assay to determine the relative amount of MUC5B mucin in samples of human cervical mucus taken through the menstrual cycle. We demonstrate a peak of MUC5B mucin in human cervical mucus collected at midcycle, compared with mucus from early or late in the cycle. This peak in MUC5B content coincides with the change in mucus character that occurs at midcycle, suggesting that this large mucin species may be important to sperm transit to the uterus.

Decidual transformation of human extrauterine mesenchymal cells is associated with the appearance of insulin-like growth factor-binding protein-1.

Human extrauterine decidual cells were studied for the presence of insulin-like growth factor-binding protein-1 (IGFBP-1) by using an avidin-biotin complex immunoperoxidase method with a purified monoclonal antibody 6303 against IGFBP-1. It is well established that human decidua expresses mRNA for IGFBP-1 and synthesizes and secretes this protein. We now describe, for the first time, that cells morphologically similar to uterine decidual cells found beneath the peritoneal mesothelium in inguinal hernia and in the cervical mucosa during normal pregnancy and in the mesenchymal tissue of the Fallopian tubes and ovaries at term react with the monoclonal antibody for IGFBP-1 equally to endometrial decidual cells. These results imply that the extrauterine mesenchymal cells, which have undergone decidual transformation, are not only morphologically but also functionally, similar to their counterparts in the uterus. Secondly, the data suggest that IGFBP-1 expression is associated with a special type(s) of cells and a certain stage of differentiation rather than special organs in the body.

Expression of blood group A and B antigens in nuclear heterochromatin in mucous cells of human cervical glands.

Using a post-embedding immunogold labeling procedure, we found that monoclonal antibody against A (MAb-A) or B antigen (MAb-B) reacted with nuclear heterochromatin regions, as well as secretory granules, in mucous cells of human cervical glands. Systematic and critical observation of specimens from 24 individuals of different blood groups revealed that the labeling pattern with MAb with strictly dependent on the blood group (A,B, or O) of the donors, i.e., MAb-A reacted with the heterochromatin from blood group A and AB but not with B and O individuals. Labeling with MAb-B was also specific for the heterochromatin from blood group B donors. On the other hand, MAb against H antigen did not react with the heterochromatin from any individuals examined, despite the fact that H antigens were detected by the MAb in secretory granules. Such specific reactions provide evidence that certain types of blood group-related antigens exist in the nuclear heterochromatin in mucous cells of human cervical glands. In contrast to the secretory granules in which ABH antigens were recognized by blood group-specific lectin, heterochromatin regions had little or no affinity for these lectins. Furthermore, the secretory status of individuals affected the staining intensity with MAb in secretory granules but not in the heterochromatin. These results suggest that the blood group substances found in the heterochromatin may have different molecular properties from those in the secretory granules, although both have the same determinant structures of ABH antigens.

Genital actinomycosis and intrauterine contraceptive devices. Cytopathologic diagnosis and clinical significance.

This study from a community hospital documents the relatively frequent occurrence of Actinomyces in 36 women diagnosed by Papanicolaou stained cervicovaginal smears. Actinomyces was identified exclusively in patients wearing an intrauterine or vaginal foreign body (intrauterine devices and pessaries). By using fluorescein isothiocyanate labeled antiserum, the organisms were shown to be Actinomyces israelii in eight randomly selected smears. In five cases Actinomyces was demonstrated in tissues obtained by endocervical and endometrial curettage. Anaerobic microbiologic studies performed in 11 of the 36 cases showed a relatively high rate of recovery of Actinomyces (36.3 per cent) with a yield of four positive cultures. In two cases the isolated organisms were shown to be Actinomyces israelii. In the remaining two cases the exact species of Actinomyces has remained uncertain. Of the 36 patients with smear evidence of Actinomyces, nine (25 per cent) had pelvic inflammatory disease, a rate that appears to be much higher than the already high rate among all intrauterine device users. This observation underscores the importance and clinical significance of the finding of Actinomyces in the vaginal smears of intrauterine device users. Papanicolaou stained cervicovaginal smears provide a relatively easy, inexpensive, fast, and highly specific method for the morphologic diagnosis of Actinomyces and allows us to recognize at a relatively early stage a group of patients who are potentially at risk for the development of pelvic inflammatory disease and its associated more severe complications. The hope is that prompt identification of Actinomyces in cervicovaginal smears can help to prevent the more serious infectious complications in intrauterine device users.

PIP: Occurrence of Actinomyces in 36 women (from September 1976-May 1977) diagnosed by Papinocolaou (Pap)-stained cervical smears (total of 9191 smears) is characterized as "relatively frequent," occurring exclusively in those women wearing an IUD or foreign vaginal body (pessaries). Fluorescein isothiocyante labeled antiserum was used to identify Actinomyces israelii in 8 randomly selected smears. In 5 cases, Actinomyces was demonstrated in tissues obtained by endocervical and endometrial curettage. The recovery rate of Actinomyces in anaerobic microbiological cultures was 36.3%, with a yield of 4 positive cultures. Of the 36 patients with smear evidence of the organism, 9 (25%) had pelvic inflammatory diseases (PID); this rate appears to be much higher than the high rate already established for IUD wearers. Pap-stained smears, easily used in a community hospital, provide an easy, inexpensive, fast and highly specific method for morphologically diagnosing Actinomyces, and its use is recommended to recognize at a relatively early stage a group of patients who are potentially at risk for the development of PID.

Ultrasonic dilatation of the uterine cervix: morphologic evaluation of a new device in the sow.

The present study demonstrates that low-power ultrasonic activation of Hegar dilators reduces the mechanical force required for dilatation of the cervix. The acute and possible late effects of an ultrasonic dilator on the cervical mucosa in the sow were evaluated by histologic study and compared to the effects of similar mechanical dilatation. In both groups early effects were mild to moderate edema and vascular congestion of the cervical mucosa. The immediate changes were comparatively more pronounced after ultrasonic dilatation. No significant pathologic differences were noted five weeks after ultrasonic dilatation of the endocervical canal. Further study may prove the device useful for dilatation of the stenosed cervix and urethra in women.

Cytologic and histologic findings in women using an IUD.

The present study was undertaken to determine the source of atypical squamous metaplastic cells that are found in large numbers in smears from women with an intrauterine contraceptive device (IUD). The cytologic data are obtained from a population screening program totaling 61,151 women, of which 1401 were using an IUD. Histologic data comprise findings from curettings on removal of an IUD and scrapes from the removed IUD in 102 women. These implicate the endometrial surface epithelium as the main source of the atypical cells in cervical smears from IUD users. As this epithelial atypia frequently disappears upon removal of the IUD, conservative follow-up of women with cytologic metaplastic atypia in cervical smears during IUD use is strongly recommended.

A new simple test for detecting rupture of the fetal membranes.

Obstetricians often have difficulty diagnosing rupture of fetal membranes because of several factors that interfere with the available tests. A new test is proposed herein, which is based on heating endocervical material on a glass slide for one minute. When the fetal membranes are ruptured, the material on the slide turns white; when they are intact, the material turns brown. The low cost, risk-free method is a simple procedure providing immediate results, thus making its use practical in daily practice.

Cytologic changes following the use of oral contraceptives.

Abnormal cytologic findings in gynecologic smears were more common among patients using oral contraceptives than among control patients. Cytologic examination should be routinely performed before oral contraceptives are prescribed. The exfoliation of endometrial cells was essentially the same between control and oral contraceptive patients. No evidence of excessive stimulation of epithelial element in endometrium accompanied the continued use of hormonal contraceptives. In general, the cytohormonal effect associated with oral contraceptives showed a depressed superficial cell count in the vaginal smears. Teenagers using oral contraceptives constituted 2 to 3% of the surveyed group. Long-term effects of antiovulatory drugs in young females are yet to be observed.

Laparoscopic recovery of sperm from peritoneal fluid, in patients with negative or poor Sims-Huhner test.

Several opinions about the importance of the Sims-Huhner test (SHT) are reported in the literature. Paradoxical are the results relative to conception following a positive or negative SHT. Pregnancy frequently occurs in women with a negative or poor SHT. This observation prompted the author to look for the presence of sperm in peritoneal fluid during laparoscopy in eight patients with a negative SHT and in three with a poor SHT. Complete evaluation was performed to rule out other infertility causes. The peritoneal fluid was aspirated from the cul-de-sac and again after washing the Douglas space and fimbriated portion of the fallopian tubes with 5 to 10 ml of normal saline solution, using a Verres needle. Samples were processed with the Sayk unit, by a special cytosedimentation technique developed for study of the cerebrospinal fluid. In six of the eight patients with a negative SHT and in the three with a poor SHT, sperm were found in the aspirate. No immature or morphologically abnormal sperm were recovered. This procedure illustrates the limitation of the long-accepted SHT in the prognosis for infertile couples. Investigations to eliminate the fixed stage in the processing are being studied to standardize the test.

The cyclical changes in the physical properties of the cervical mucus and the results of the postcoital test.

PIP: The effect of the postcoital test on the cyclic changes in the physi cal properties of the cervical mucus during the preovulatory period was investigated, and the results obtained in women who conceived during the period of investigation (fertile group) were compared with the results obtained in those who did not conceive (infertile group). 3007 postcoital examinations were performed within 72 hours after intercourse on 505 couples complaining of infertility. No significant differences were found in the amount, viscosity, and spinnbarkheit of the cervical m ucus or in the motility and drive of spermatozoa between 94 couples who conceived and 411 couples who remainded infertile. The number of spermatozoa per high power field was significantly higher in the fertile group as compared with the infertile when the amount of mucus increased. When estrogenic changes in the mucus first became apparent, the concentration of spermatozoa rose and their motility and drive improved. No further improvement was seen with further changes in the mucus. When white blood cells in the mucus appeared, both the motility and the drive of spermatozoa decreased significantly, but their number per high power field did not change. No significant changes were found to occur in either motility or drive during 72 hours after intercourse and in the number of spermatozoa per high power field during the first 24 hours. However, concentration of spermatozoa decreased significantly during Days 2 and 3. In 26 tests following the coitus presumably responsible for pregnancy, the concentration, motility, and drive of spermatozoa in mucus were not found to be different from those for the whole group of fertile patients. The wide range of distribution of the results in both groups indicates that the results of the postcoital test cannot be used as a reliable index of fertility in an individual couple.^ieng

Morphology of postcoital spermatozoa in the cervical secretion and its clinical significance.

Morphologic analysis of spermatozoa recovered 10 hours postcoitally from the human female genital tract and its correlation to the clinical situation have shown that (1) spermatozoa in the vagina are equivalent to those in the semen sample; (2) spermatozoa within the cervical secretion exhibit significantly better morphology than do those of the semen sample, indicating the presence of a barrier against abnormal spermatozoa at the level of the external os; (3) this barrier is significantly more effective in cases where conceptions later occur than in cases of persistent, unexplained infertility; and (4) the barrier is particularly active against spermatozoa with abnormal heads.