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Contamination of embryo transfer catheter and treatment outcome in in vitro fertilization.

PURPOSE: The effect of embryo transfer (ET) catheter contamination with mucus and/or blood on treatment outcome in an in vitro fertilization (IVF) program was evaluated. METHODS: One thousand four ET procedures in 877 patients having conventional IVF in the long (1189 cycle) and short (15 cycles) protocol were analyzed to determine the impact of catheter contamination on the incidence of retained embryos and the clinical pregnancy rate. RESULTS: Catheter contamination with mucus and/or blood is a feature of difficult ET. Embryos were significantly more likely to be retained when the transfer catheter was contaminated with mucus (17.8 versus 3.3%) or blood (12 versus 3.3%). When the catheter was contaminated and an embryo(s) was retained and immediately retransferred, the pregnancy rate was not compromised. The pregnancy rate was significantly reduced when the ET catheter was contaminated with blood (15.5 versus 27.1%; P = 0.002), but no embryo was retained in the catheter set. CONCLUSIONS: Catheter contamination compromises the treatment outcome in IVF only when there is no associated retained embryo(s). As increased vigilance in searching for extruded embryos may not be practical, we suggest that cervical mucus should be routinely aspirated and ET performed as atraumatically as possible.

Adult↗

A simple method of producing a monolayer of cervical cells for digital image processing.

A manual method of producing a monolayer of isolated and evenly distributed cervical epithelial cells has been developed for use in an automated digital image analysis system. The cervical scrape specimen is vortexed from the spatula into a solution of Mucosol. The suspension is filtered through a 10 micro nylon mesh to remove polys and debris and then syringed to disaggregate epithelial clumps. The cells are plated out on a Nuclepore filter. The filter is touched directly onto a glass microscopic slide to transfer cells, which are then fixed and stained in a routine manner. Cell counts reveal elimination of 92% polys, with only 2% loss of squamous cells.

Autoanalysis↗

Immunoglobulin levels in the serum and cervical mucus of tailed copper IUD users.

Immunoglobulin levels were measured by agar gel single radial immunodiffusion in the serum and cervical mucus of 50 tailed copper IUD users, 20 combined oral contraceptive users and 20 women not using contraception (controls). IgG, IgA and IgM levels were significantly higher in the copper IUD users compared to the other two groups. It is not known whether this was related to the small numbers of bacteria found in the uterine cavity of tailed IUD users or to the foreign body reaction of the device.

Adult↗

Effect of Lippes loop on sperm recovery from human fallopian tubes.

Recovery of human sperm from the fallopian tubes after natural insemination have been studied in sixty women; out of these, thirty cases were fitted with the Lippes loop, type C, for at least six months. The mean sperm count/high power field (HPF X 400) and percentage of motile sperm in the cervical mucus in both control and Lippes loop groups showed no significant differences. No sperm were recovered from the fallopian tubes of the Lippes loop group, while fourteen control cases had a positive recovery of sperm from the tubal washings. The mean number of white blood corpuscles in the tubal washings of Lippes loop-fitted cases was found to be significantly higher than in control cases. The phagocytic capacity of these cells may explain the low incidence of sperm recovered from the fallopian tubes in such cases.

Adult↗

Fertility awareness-based methods for contraception: systematic review of randomized controlled trials.

CONTEXT: Fertility awareness-based (FAB) methods of family planning estimate the fertile days of the menstrual cycle whether by observing fertility signs such as cervical secretions and basal body temperature or by monitoring cycle days. The efficacy and acceptability of these methods are unclear. METHODS: We retrieved and analyzed all randomized controlled trials that examined any FAB method used for contraception. RESULTS: Because of poor methods and reporting, pregnancy rates could not be determined. A trial in Colombia found similar numbers of pregnancies among women randomized to the ovulation and symptothermal methods, but a trial in Los Angeles observed more pregnancies in the group assigned to the ovulation method. In the two American trials, recruitment of participants was unexpectedly difficult. Continuation rates were poor. CONCLUSION: Despite intensive training and ongoing support, most participants in these trials discontinued prematurely. The comparative efficacy of these methods remains unknown. However, with the ovulation and symptothermal methods, pregnancies appear to be common; method continuation rates are low.

Awareness↗

Assessment of the anti-microbial agent C31G as a spermicide: comparison with nonoxynol-9.

The broad-spectrum anti-microbial agent, C31G, containing an equimolar mixture of n-dodecyl-dimethylamine-N-oxide (C12-N-O) and N-(n-dodecyl), N-dimethyl-glycine (C12-betaine), was tested for spermicidal activity in comparison with the currently used spermicide, nonoxynol-9 (N-9). The rate of sperm cell permeabilization by the spermicides, as assayed with the fluorescent probe, TO-PRO-1, increased as the cube of the C31G concentration, while the rate increase was linear with N-9 concentration. At 0.04%, the rate of sperm cell permeabilization with both spermicides is at the limit of rapid measurement. C31G diffuses through cervical mucus at a more rapid rate than does N-9. C31G has long been known to aid wound healing and reduce inflammation, whereas N-9 has been reported to induce vaginal irritation. C31G would, thus, seem to have the spermicidal efficacy, the broad range of anti-microbial activity, and the lack of inflammatory activity that is sought in the ideal vaginal spermicide.

Adult↗

Biological activity assessment of a novel contraceptive antimicrobial agent.

Microbicides are a new category of compounds being developed as a prophylactic approach for the prevention of transmission of sexually transmitted diseases (STDs), including the human immunodeficiency virus (HIV). These are primarily being developed as women-controlled methods, with the target of designing new compounds or formulations that can be used without the knowledge of a male partner. Microbicide screening can be initially based on their hyaluronidase-inhibiting (HI) activity, as this enzyme plays a major role in the sperm and microbe penetration into the substrate. Derivatives of hesperidin, a citrus flavonoid glycoside, have been reported in the literature for their HI effects. Hesperidin was thereby sulphonated under strictly controlled conditions and the active fraction isolated and characterized, based on its HI activity. This derivative was screened for antimicrobial and enzyme-inhibitory activities, specifically for the reproductive tract. Sulphonated hesperidin (SH) was found to completely inhibit the sperm enzymes hyaluronidase, giving an indication toward its contraceptive effects. It was also been found to inhibit various sexually transmitted pathogens, including Chlamydia trachomatis, Neisseria gonorrhoea, HIV, and Herpes Simplex virus type 2 (HSV-2). Its safety assessment was based on its noninterference in sperm motility and its penetration through the cervical mucus, and no effect on the growth of lactobacilli, the normal vaginal flora. It was also found to be nontoxic to the HIV substrate cells (MT2 cells). The study concludes that sulphonated hesperidin can be developed as a potential microbicide for a dual prophylaxis of contraception and transmission of STDs and AIDS.

Anti-Bacterial Agents↗

Critical evaluation of methylcellulose as an alternative medium in sperm migration tests.

BACKGROUND: The aim of this study was to evaluate the ability of human spermatozoa to penetrate methylcellulose (MC) and to compare this with penetration in hyaluronic acid. METHODS: Spermatozoa from normal (>or=20 x 10(6) sperm/ml, >or=50% progressive motility, >or=5% normal forms) and oligozoospermic (<20 x 10(6) sperm/ml) semen samples were allowed to swim into glass capillary tubes containing methylcellulose with a viscosity of 15 centipoise (cp) (MC15) and 4000 cp (MC4000), hyaluronic acid (rooster comb) or Sperm Select. Penetration of the spermatozoa at 1, 2, 3 and 4 cm were correlated with basic semen parameters (concentration, motility and morphology). The effects of temperature on penetration into MC4000 were explored at 17-37 degrees C. RESULTS: Higher numbers of spermatozoa penetrated MC4000 (10 mg/ml) compared with MC15 and the hyaluronic acid preparations. There was a highly significant correlation between the number of spermatozoa at all migration distances in MC4000 (10 mg/ml) and semen parameters. Increases in temperature from 17-37 degrees C were accompanied by significantly higher numbers of spermatozoa at each penetration distance. MC4000 at 10 mg/ml was at least as favourable to sperm penetration as human cervical mucus. Effective discrimination between normal and abnormal samples was achieved using MC4000 (10 mg/ml). CONCLUSION: Our results suggest the potential use of methylcellulose (MC4000, 10 mg/ml) as a reproducible and effective alternative to hyaluronic acid in sperm migration tests.

Cervix Mucus↗

Correlation between seminal carnitine and functional spermatozoal characteristics in men with semen dysfunction of various origins.

BACKGROUND AND OBJECTIVE: L-carnitine is an essential molecule involved in mitochondrial metabolism, controlling the transport of acetyl and acyl groups across the mitochondrial inner membrane. Carnitine and acetylated carnitine (L-acetylcarnitine) are found in high concentrations in the epididymis, where they also act as antioxidants, protecting spermatozoa against damage caused by reactive oxygen species. In this open study we investigated the correlation between seminal carnitine levels and spermatozoal function, and the effect of combined L-carnitine + L-acetylcarnitine therapy, in infertile men. PATIENTS AND METHODS: 170 infertile men were enrolled in this study. Patients were divided into those with a total sperm motility below the normal WHO range (<50% motility, group 1 [n = 102]) and those with total sperm motility within the normal range (> or =50% motility, group 2 [n = 68]). Patients in group 1 were further divided into two groups: those with primary or secondary azoospermia (1B [n = 36]), and those without (1A [n = 66]). Patients in group 1A received L-carnitine 1 g/day and L-acetylcarnitine 500 mg twice daily for 6 months. Seminal carnitine levels were measured and correlated with sperm count and motility, eosin test, hypo-osmotic swelling test, acridine orange test for sperm nuclear DNA integrity and sperm kinetics evaluated by computer-assisted sperm analysis in all patients. RESULTS: There was a significant correlation between seminal carnitine concentration and sperm concentration, total sperm count, sperm total motility, rapid forward progression, live sperm count, membrane function, nuclear DNA integrity, capacity for cervical mucus penetration, linearity of spermatic movement, and amplitude of lateral sperm head movement (all p < 0.0001) in the entire study population. In group 1A, there was a significant increase in total motility, live sperm count, membrane integrity and linearity of spermatic movement after 3 and 6 months of L-carnitine/L-acetylcarnitine treatment, and in capacity for cervical mucus penetration after 6 months of treatment, compared with baseline. CONCLUSION: Seminal carnitine concentration may be an appropriate marker of sperm and epididymal function. L-carnitine/L-acetylcarnitine treatment may be an effective therapy to improve mainly functional seminal parameters.

Acetylcarnitine↗

Self-sampled and air-dried cervicovaginal secretions can be used for analyses of mucosal antibodies.

Cervicovaginal secretions were collected from 26 women (13 premenopausal and 13 postmenopausal) using a new sampling device (MucoSafeTM) with an absorbent which was introduced into the vagina and retrieved by the women themselves, after which it was air-dried and stored for months at room temperature until extraction of immunoglobulins. Cervical secretions were also collected by absorbent cylindrical wicks (Polyfiltronics) which were introduced into the cervical canal during speculum examination and thereafter kept frozen until extraction. The concentrations of specific IgA and IgG antibodies (to group B streptococci) in extracts from both methods were corrected by reference to total immunoglobulin levels. Three pairs of samples, all from postmenopausal women, were excluded from analysis due to undetectable levels of antibodies in the MucoSafeTM specimen. In the remaining 23 pairs, corrected concentrations of IgA and IgG antibodies in samples obtained by MucoSafeTM correlated well with the corresponding concentrations in wick samples, R = 0.84 (p < 0.0001) and R = 0.69 (p = 0.0002), respectively. Thus, cervicovaginal secretions for antibody measurements can be obtained by this novel method for self-sampling, obviating the need for speculum examination and storage of frozen samples.

Absorption↗

Spinability of bronchial mucus. Relationship with viscoelasticity and mucous transport properties.

Spinability of normal mucus and of sputum collected in chronic bronchitic patients was measured using an automatic device derived from that developed by CHRETIEN et al (1977) for cervical mucus. Spinability of sputum decreased as the purulence increased. Although significant correlations were found between spinability and apparent viscosity (r = - 0.50, p less than 0.05) or elasticity (r = 0.54, p less than 0.02), large variations in spinability were observed for sputum samples ranging in a zone of low viscosity and elasticity. Sputum with high spinability exhibited normal transport rate on the depleted frog palate although their viscosity and elasticity were abnormally low. The significant correlation obtained between spinability and sputum transport rate (r = 0.61, p less than 0.01) suggests that this rheological factor, along with viscosity and elasticity, plays an important role in the mucociliary transport mechanism.

Adult↗

Release characteristics, ovarian activity and menstrual bleeding pattern with a single contraceptive implant releasing 3-ketodesogestrel.

The properties of a single contraceptive subdermal implant releasing 3-ketodesogestrel were assessed in fifteen women over twelve months. Serum levels of 3-ketodesogestrel were monitored regularly following insertion and after removal. The mean serum level of 3-ketodesogestrel was 245 pg/ml after 72 h (steady state) and 176 pg/ml after twelve months. All volunteers demonstrated ovulation inhibition throughout the study. Transient oestradiol peaks occurred during the study. No luteal activity was noted. The cervical mucus was rapidly rendered hostile to sperm migration. Two women withdrew from the study during the first six months for medical reasons. Both volunteers cited bleeding irregularity as the main cause, one complaining of oligomenorrhoea, the other of prolonged bleeding/spotting episodes. A small but significant increase in weight was noted during the study period.

Adolescent↗

The in vivo biodegradation of nylon 6 utilized in a particular IUD.

The particular IUD of this study was composed of a plastic shield and a nylon 6 coated filament tail string. This IUD was unique, in the fact that its tail string was the only one (and has been to date) that consisted of a coated multifilament arrangement. Studies on the tail string have revealed bacteria and debris existing between the outer filaments that are underneath the coating. Other studies have reported that removed devices were found to exhibit an unusually high incidence of breaks in the tail string coating. Studies at the University of Lowell also found that upon examination of devices that had been in vivo for periods of two years or longer, 67% of these devices examined had breaks in the coating of the tail string. Investigations have determined that the design of the tail string was serving as an avenue of transport for the bacteria to bypass the protective cervical mucus barrier and enter the sterile uterine cavity through the breaks in the tail string coating. This mechanism has been suggested to account for the higher frequency of uterine infections associated with this device. Research at the University of Lowell was undertaken to study the degradation of the nylon 6 coating and to determine the in vivo degradation mechanism. The biochemistry of the fluids present in the reproductive tract were carefully documented and the more aggressive ones selected for in vitro trials. The results of the study revealed that the in vivo physical breakdown of the coating may depend upon a series of mechanisms and variables.

Biodegradation, Environmental↗

Diagnosis of malignant tumors on the basis of the current generating capacity of malignant tumorous tissue. II. Study of the malignant tumors of female genital organs.

Description of an electrochemical measuring method for the in vivo quick-diagnosis of malignant tumors and dysplastic alterations of the cervical epithelium is given. The measuring results carried out in 660 cases on the surface of the cervical epithelium are reported. The current generating capacity of planocellular carcinoma of the cervical epithelium exceeds the upper limit of the normal value with 40% and is significantly higher than the values measured in dysplastic areas. With the aid of measuring, dysplastic alteration are demonstrable even in cases, in which the cytologic or coloposcopic examination is not indicative of dysplasia. Measurings in the benign and malignant tumorous tissue were carried out with needle electrode in 41 cases, during operation. The current generating capacity of malignant tumors exceeded with 150--170% the values measured in the healthy tissue, respectively in benign tumorous alterations.

Cervix Mucus↗

Thionine-Feulgen Congo Red -- a new staining technique for automated cytology.

A new staining method has been developed for an automated pre-screening device for cervical smears called BioPEPR. The method is a combination of the Feulgen technique with thionine as th Schiff reagent, and congo red. Thionine and congo red have absorption maxima at 585 nm and 500 nm respectively. The staining results in cells with an orange-red cytoplasm and brownish-blue nucleus. These smears can e easily screened using a normal microscope. The BioPEPR system scans photonegatives of the smears that are made at a magnification of ten using monochromatic light of 545 nm. The staining facilitates good discrimination between background and cytoplasm and between cytoplasm and nucleus, allowing morphologic parameters such as cytoplasmic area, nuclear area and nuclear integrated darkness to be well measured.

Autoanalysis↗

Regulation of the paracellular permeability of cultured human cervical epithelium by a nucleotide receptor.

OBJECTIVE: We sought to determine what type of regulation of transepithelial transport in leaky epithelium can occur at the level of paracellular permeability. METHODS: The epithelial permeability to the polar acid pyranine (Ppyr) and the transepithelial electrical resistance (R) were determined in cultured human cervical epithelium. RESULTS: Extracellular adenosine triphosphate (ATP) acutely and reversibly decreased the paracellular permeability, as evidenced by an increase of R from 10 +/- 3 to 16 +/- 2 omega/cm2 and a decrease in Ppyr from 13 +/- 3 to 9 +/- 3 x 10(-6) cm/sec and (P < .01 for both). The ATP effect was dose related (average median effective concentration 2 +/- 1 microM), saturable at 50 microM, and desensitized with repeated administration; it was mimicked by uridine triphosphate and ATP-gamma-S but not by adenosine monophosphate, adenine, adenosine, or adenosine diphosphate. The ATP effect on resistance remained intact even when the intercellular resistance was decreased with a basal-to-apical pressure gradient, but was abolished by lowering extracellular calcium. CONCLUSIONS: These results indicate the following: 1) Paracellular permeability in the cervical epithelial cell line is regulated by a nucleotide receptor; and 2) the tight junctions are effectors of nucleotide-receptor stimulation. We suggest that extracellular ATP may regulate mucus production in the cervix in vivo by acting on a surface receptor and by increasing the resistance of the tight junctions.

Adenine↗

Implantable contraception.

Although levonorgestrel contraceptive implants have been available for over 15 years, innovations have only recently led to a wider choice. These new implants offer easier insertion and removal and other advantages depending on the type of progestin. Implants prevent pregnancy by several mechanisms, including inhibition of ovulation and luteal function and alteration of cervical mucus and the endometrium. The high efficacy and ease of maintenance make implants an ideal contraceptive for many women, including adolescents, a population that uses implants infrequently but reports high satisfaction. Implants are appropriate for women who are breastfeeding, who have contraindications to estrogen, or who have diseases such as diabetes, hypertension, sickle cell anemia, or an HIV infection because implants have few metabolic or hematologic effects. Long-term use has not been associated with a decrease in BMD and generally leads to increased blood levels and iron stores. Women who wish to space their pregnancies appreciate the nearly immediate onset of action with insertion and the rapid termination of all effects with removal. All types of implants lead to menstrual changes and other side effects in some women. Adverse effects that occur in implant users more than the general population include headaches and acne. Women must be thoroughly counseled regarding the potential for menstrual alteration, side effects, and sexually transmitted infections if they do not use condoms. Despite their initial high cost, implants are a cost-effective method over several years, even when discontinued before the life of the implant.

Adolescent↗

Response of midpiece vesicles on human sperm to osmotic stress.

BACKGROUND: We investigated the osmotic response of midpiece vesicles (MPV) on human sperm. METHODS: Light microscopy, transmission X-ray microscopy and computer-aided semen analysis was used to investigate sperm in normozoospermic semen from healthy donors, separated from semen and suspended in hyper- or hypo-osmotic solutions. RESULTS: The mean incidence of MPV-bearing sperm in semen was 31% (range 8-71; n = 47). MPV were morphologically different from cytoplasmic droplets. The incidence of MPV-bearing sperm in separated populations increased reversibly after washing in Sperm Preparation Medium but not after washing in seminal plasma. There was an inverse relationship between medium osmolality and both MPV-bearing sperm incidence and MPV diameter. However, initial osmolality in semen from different donors did not correlate with incidence of MPV-bearing sperm. Furthermore, a direct relationship was observed in semen as osmolality increased with time. No correlation existed between progressive motility and semen osmolality. Progressive motility and the amplitude of lateral head displacement were significantly reduced in sperm with an MPV (three out of four semen samples, 26-32 sperm). The incidence of MPV-bearing sperm in those that had penetrated cervical mucus (75, 46, and 40%) was increased compared with the adjacent semen (24, 35, and 24%). CONCLUSIONS: MPV are ubiquitous and distinct from cytoplasmic droplets. They respond to osmolality changes of the surrounding medium. The presence of an MPV can reduce motility but not survival in cervical mucus. Therefore, they should not be considered detrimental to sperm function.

Cervix Mucus↗