Epididymal stereocilia in semen of infertile men: evidence of chronic epididymitis?

As free stereocilia in semen samples from infertile men could be related to a subclinical chronic infection of the seminal tract, we carried out a retrospective analysis of 14 files of infertile men with stereocilia in semen (group 1) and in two control groups: 30 infertile men from the same study population (group 2) and 40 fertile sperm donors (group 3). Clinical, bacteriological, sonographic and microscopic findings associated with these semen samples were compared. Epididymal cysts were found in two of 14 men in group 1 and in four of 28 in group 2. Positive semen cultures for Chlamydia trachomatis, Mycoplasma hominis or Ureaplasma urealyticum (chi(2) = 8.20; P = 0.002; OR: 7.22) and varicocele (chi(2) = 8.96; P = 0.002; OR: 2.25) were associated to the presence of free stereocilia in semen. These data suggest that a silent chronic infection of the epididymis by C. trachomatis, M. hominis, U. urealyticum and/or inflammation produced by varicocele might be the cause of stereocilia loss in the semen.

Sperm dysfunction in subfertile patients with varicocele and marginal semen analysis.

A comparative analysis of potentially functional spermatozoa per ejaculate, progressive motility, hypo-osmotic swelling test, acrosome integrity and sperm viability (24 and 48 h) was carried out in a group of 40 subfertile patients with varicocele and marginal semen analysis and 40 fertile subjects, in order to identify subclinical abnormalities that may explain subfertility. Patients with varicocele had lower numbers of potentially functional spermatozoa per ejaculate, progressive motility, acrosome and membrane integrity and sperm viability. These abnormalities were not related to the grade of varicocele, testicular volume or serum FSH concentration. A positive correlation between the hypo-osmotic swelling test and progressive motility (r = 0.71) and between potentially functional spermatozoa and the hypo-osmotic swelling test (r = 0.69) was found in patients with varicocele. These data suggest that some of the deleterious effects produced by the varicocele might be related to sperm migration and viability in the female genital tract and others to sperm-zona interaction and/or sperm-egg fusion.

Bacteriospermia and male infertility: a method for increasing the sensitivity of semen culture.

OBJECTIVE: To test a method for increasing the sensitivity of semen culture. Design-Prospective and transverse. Setting-Andrology clinic at a tertiary care health institution in Mexico City. PATIENTS: 65 infertile patients with abnormal semen, bacteriospermia detected on Gram stain and at least two previous negative semen cultures (<3 months) were included to test routine semen culture and a method including centrifugation of semen at 10,00 rpm for 20 minutes. Localization cultures were also carried out in all patients. MAIN OUTCOME MEASURE: Bacterial isolation in semen samples. RESULTS: Routine semen culture was positive in 22% of patients, while centrifuged aliquots of the same semen sample were positive in 52% of patients (chi2 = 6.60, P < .01). Enterococcus was isolated in 43% of patients, E. coli in 24%, coagulase-negative Staphylococcus in 19%, and U. urealyticum in 14%. Ninety percent of isolates corresponded to specimens from the urethra and the prostato-vesicular region. CONCLUSION: Sensitivity of semen culture increased with centrifugation of semen samples. Localization pattern and type of isolates suggest that these patients had chronic prostatitis and that episodic elimination of bacteria might also explain false negative semen cultures in patients with chronic asymptomatic infection of the accessory sex glands.

[Epidemiologic characteristics in women with sterility and cervical lesions with and without human papilloma virus infection. Comparative study]. / Características epidemiológicas en mujeres con esterilidad y lesiones cervicales con y sin infección por virus de papiloma humano (VPH). Estudio comparativo.

Human papilloma virus (HPV) infections frequently cause cervical lesions of different morphologies. We have previously reported a 53.5% pregnancy rate after treatment in a group of women with infertility associated to HPV infection of the cervix uteri. In that paper it was stated that a controlled study should be conducted in order to confirm this finding. Present work was aimed to find if there is any correlation between HPV infections of the cervix uteri and infertility in a retrospective design using an historical cohort of patients studied between 1991 and 1996 in our clinic. A total of 61 women attending the Infertility Clinic at the Instituto Nacional de Perinatología were included into two group. Group 1 (n = 45) included women with HPV lesions of the cervix and group II (n = 16) was formed by women with other type of cervical lesions who had no evidence of HPV infections on colposcopy. Cervico-vaginal citology, colposcopyc study and biopsy specimens were evaluated in all this medical records and the patients status (pregnant-not pregnant) at one year after treatment was registered. The mean duration time of infertility was 4.86 in group I and 3.5 in group II. Pregnancy rate was 16/45 (35.55%) in group I and 6/16 (37.5%) in group II. Seventy five percent of patients in group I and 66% in group II achieved a spontaneous pregnancy after specific treatment of cervical lesions whereas 25% and 33.3% required only ovarian stimulation with clomiphene or hMG. Pregnancies occurred approximately at 9 months after treatment in group I and at 7 months in group II. An association of cervical lesions and a tuboperitoneal factor (excluding endometriosis) was found on 53.57% of women in group I and on 46.66% of women in group II. Cervico-vaginal cytology was suspicious of HPV infection in less than 25% of cases. Present study emphasizes the need for a colposcopic study for the diagnosis of HPV infection in infertile women with cervical lesions even in cases with a negative cervico-vaginal cytology; because specific treatment of these lesions may yield spontaneous pregnancies. It also demonstrates that around 50% of patients with cervical lesions have an associated tuboperitoneal factor, which indicates that it is mandatory to study the possible participation of viral infections on tubal pathology.

Progesterone induces human sperm chemotaxis.

OBJECTIVE: To characterize the molecular nature of the chemotactic signal for sperm contained in human follicular fluid (FF). DESIGN: Follicular fluid was fractionated and several procedures were followed to the physicochemical initial characterization of sperm chemotactic compound(s). MAIN OUTCOME MEASURE: Relative chemotactic activity of each fraction was measured in a double chamber device. RESULTS: Sperm chemotaxis was found to be associated with a lipid-like molecule extracted from FF. Several steroids were assayed individually and only P showed sperm chemotactic properties in dose-response curves. CONCLUSIONS: In this paper we present experimental evidence to support the hypothesis that P, the main steroid component of FF, is a mediator of sperm chemoattraction in human beings.

[Chemotactic factor for spermatozoa: a new biological function of progesterone]. / Factor quimiotáctico para espermatozoides: una nueva función biológica de la progesterona.

Our group has recently described the existence of a chemoattractant factor for spermatozoa contained in the mature follicles fluid. Simultaneously it was possible to develop a new method that permits to evaluate the chemotactic capacity of spermatozoa and that due to its simplicity makes possible the systematic study of CFS features. This study considered CFS molecular characterization from follicular fluid (FF). The FF of women was studied in an Assisted Fertilization Program, that were qualified as mature according to different criteria. The FF were fractioned with different techniques that permitted to separate an active fraction with lipid physicochemical characteristics. The fine layer chromatography showed the presence of different steroids, that were individually assayed for chemotactic activity. Only progesterone showed that activity and its effect showed a dose-response curve within physiological values. Our study permitted to identify progesterone as CFS previously described. This steroid's function is rather new and its action mechanism is being studied in our laboratory.

[The clinical study of male infertility]. / Estudio clínico de la infertilidad masculina.

Male infertility study requires systematization of clinical and paraclinical diagnostic procedures and the knowledge of frontier techniques to analyze functional capacity of human spermatozoon. The application of the various methods of assisted reproduction in male infertility treatment, should be judged with a good criterion in order to offer maximal benefit and least risk. New methods to handle gametes and standardization of existing ones, could, in the future, offer better alternatives in male infertility treatment.

[The effect of the early administration of progesterone on egg recovery, fertilization and pregnancy in an institutional fertilization in vitro program]. / Efecto de la administración temprana de progesterona sobre la tasa de captura ovular, fertilización y embarazo en un programa institucional de fertilización in vitro.

39 patients which underwent an IVF-ET procedure at the INPer were divided in two groups. To one of them, progesterone was administered since the day of oocyte retrieval (group I), whereas the other group received progesterone 48 hours before hCG administration (group II). There were no statistical differences between the groups in the parameters analysed before the oocyte retrieval. There were statistical differences between the groups in progesterone levels in the day of oocyte retrieval in favour of group II, and in fertilization rate and transfer rate for group I. There were no valuable differences between the groups in pregnancy rate.

Identification and partial characterization of an immunoglobulin A protease associated with human sperm.

The ability of human sperm to evade local humoral immune response at the female genital tract has received little attention. As some microorganisms inactivate secretory immunoglobulin A (S-IgA) by enzymatic degradation as a part of their pathogenic mechanism, we decided to test the hypothesis that there is an IgA protease in human semen that helps sperm to progress through the cervix uteri. Incubation of different semen fractions in the presence of S-IgA revealed IgA protease activity associated with raw semen or sperm but not with seminal plasma; raw semen and sperm produced two split products of 66 kDa and 45 kDa, respectively, in SDS-PAGE. The enzymatic activity was blocked with EDTA. IgG and IgM were not digested under the same experimental conditions. Degradation of S-IgA by an IgA protease on the sperm surface may be among the mechanisms present in semen that enable sperm to evade an immune response in the female genital tract.

[Index of cells obtainable from semen: indicator of fertility?]. / Indice de células recuperables del semen: indicador de fertilidad?

It has been recognized that semen analysis is not a sensible nor a specific tool for detecting infertility in the male. Its predictive value is limited by the subjectivity of manual analysis, a high variability of semen parameters in fertile men and lack of correlation between sperm characteristics and fertility indexes. In this paper we present evidence supporting the concept that calculating an index of potentially fertile cells from data obtained in a regular semen analysis might be useful for predicting fertility in the male.

[Spontaneous pregnancy, independent of treatment, in sterile couples]. / Embarazo espontáneo independiente de tratamiento entre parejas estériles.

The spontaneous pregnancy independent of treatment in sterility is a frequently observed event in this group of couples. The spontaneous pregnancy appears up to 61% of women with antecedents of sterility and in selected populations of healthy women; a third of them experience, once in their lives a subfertility episode. The characterization of this event in a population with sterility, will help to define the capacity of different diagnostic methods, as well as therapeutic methods in terms of efficacy. The objective of this study was to identify women with diagnosis of sterility and spontaneous pregnancy; to describe the main characteristics: clinical, of laboratory, of gabinet, and to discuss some implications of biological variability useful in the interpretation of these tests. One hundred and eleven patients with sterility and spontaneous pregnancy, in a descriptive and retrospective design, were analyzed. Operational definitions for the main factors related with sterility, were used, as tuboperitoneal, endocrine-ovarian, cervico-vaginal and masculine. The pregnant patients related to any type of treatment, were excluded. Two study groups were established: Group I (n-46) and Group II (n = 65) with primary sterility and secondary sterility, respectively. Average age for both groups was similar (mean = 29 years old). The time of sterility was 46.52 and 43.52 months, for Group I and II, respectively. The time of pregnancy from the point zero (admission), was, in average 6.21 and 4.9 months for Group I and II. The following factors were identified as abnormal: endocrine-ovarian 28.60 by menstrual pattern, progesterone and endometrial biopsy: tuboperitoneal 12.67% by hysterosalpingography and laparoscopy; masculine 12.67 by direct spermatobioscopy.(ABSTRACT TRUNCATED AT 250 WORDS)

Novel model for study of human sperm chemotaxis.

OBJECTIVE: To develop and test an adequate method for the study of sperm chemotaxis. DESIGN: A model based on a double-chamber design and a matrix of 0.5% hyaluronic acid was used to test the response of human sperm to chemotactic signals from follicular fluid (FF). RESULTS: A plateau of migration was reached at 10 minutes of incubation; under this condition, 70% of sperm migrated toward chambers with FF, less than 20% to the opposite side, and the rest remained at the site of application. Intra-assay and interassay coefficients of variation using pools of FFs were 17.2% and 23%, respectively. CONCLUSION: These data confirm previous findings and support the use of this novel model for the study of chemotaxis on human gametes.

[Glucose determination as prognosis index of intra-amniotic infection]. / Determinación de glucosa como indice pronóstico de infección intraamniótica.

Intraamniotic infection is a frequent problem in Obstetrics, and is related with an important maternal and fetal impact, being important pre-term delivery and premature rupture of membranes. The "golden" test for this entity is bacteriological culture. Its use is limited in function of time (more than two days) and disponibility. The rapid diagnosis of infection in vital to start antimicrobial management and evaluation of uterine evacuation. Low concentrations of glucose (G) have been used as prognostic of infection in different biological compartments. The objective of this study is to evaluate the usefulness of G as prognostic index of intraamniotic infection (PIIAI) as compared with Gram tincture (GT) and bacteriological culture. Sixty four patients were included. Group I (n = 33) with infection, and group II (n = 31) without infection. Average of G for group I was 19.96 +/- 07.61 ES and 114.46 +/- 20.09 ES for the group II, with p less than 0.001. The sensitivity (S), specificity (Sp), positive predictive value (+PV) and negative (-PV) for a concentration of G in amniotic fluid less than 15 mg/dl was 72, 77, 77 and 72% respectively. The S, Sp +PV and -PV for G minor than 10 mg/dl was 69, 87, 85 and 73%. Gram tinction had a S, Sp +PV and -PV of 57, 83, 79, 65%. If both determinations are put together (G and GT), one sees and S of 88%, Sp 77% +PV 80% and -PV 85%.(ABSTRACT TRUNCATED AT 250 WORDS)

[Identification and initial characterization of a protease of immunoglobulin A associated with human spermatozoa]. / Identificación y caracterización inicial de una proteasa de inmunoglobulina A asociada a espermatozoides humanos.

The factors that the human semen use to block the humoral immune response at the uterine cervix level are not known. This response is mainly represented by immunoglobulin A, secretory type (IgAs). A possible mechanism would be hydrolisis od IgAs in an analogue ways as it has been described for some pathogenous bacteria that colonized human epitheliums and hydrolize IgAs. The objective of this study was to search for the presence of a protease for IgAs in human semen, seminal plasma and espermatozoons, using immunoelectrophoresis in agarose and electrophoresis in polyachrilamide, doing after immunotransference in nitrocellulose membrane. It was possible to identify a protease in sepermatic fraction, that hydrolizes IgAs, producing two fragments, the first one with molecular weight 66 to 70 kD, the degrees C. It is a metaloprotease (inhibited by EDTA). Its acti-enzymatic activity was optimal at 24 hours, pH 7.5 at 37 C. It is a metaloprotease (inhibited by EDTA). Its activity was specific for IgAs, and do not affected other immunoglobulins (IgG and IgM). The enzymatic activity of spermatozoon demonstrated in this study, may be on of the necessary factors used for blocking humoral immune response in femenine genital tract, and to permit that spermatozoon achieve the fecundation of human ovocyte.

Bacterial vaginosis associated with G vaginallis / Mobiluncus sp: ultrastructural parameters.

PIP: Physicians at the National Institute of Perinatology in Mexico City, Mexico used a Carl-Zeiss EM 10C electron microscope to examine genital secretion samples from 10 pregnant women (15-38 weeks' gestation) who had been diagnosed with Mobiluncus species and Gardnerella vaginalis infections to illustrate the form and structure of bacteria responsible for bacterial vaginosis. They were concerned that these bacteria induce preterm labor and premature rupture of membranes (PL/PROM). These bacteria have been present in the genital tract of 30% of pregnant women with a thick whitish discharge who have attended the Institute's prenatal outpatient clinic. Physicians noted on the microscope slides that bacteria surrounded vaginal squamous epithelial cells (clue cells). Numerous gardnerella-like bacteria surrounded elongated squamous epithelial cells with many plasma projections. An extensive area of lysis existed around the bacteria in the cytoplasm of many squamous epithelial cells with intact membrane and nonexistent microfilaments. This finding indicated that the bacteria invade and destroy the cells. Plasma membrane projections almost completely surrounded the gardnerella-like bacteria in certain areas. Since this study strengthened the theory that G. vaginalis enters the vaginal squamous epithelial cells, researchers should conduct more studies to determine its role in PL/PROM.^ieng

Human amniotic fluid modulation of collagenase production in cultured fibroblasts. A model of fetal membrane rupture.

The participation of a mechanical factor as the only cause of rupture of fetal membranes during normal labor or premature rupture has been criticized, and the involvement of an enzymatic mechanism has been proposed. In this study we analyzed the effect of human amniotic fluids at different gestational ages on the collagenase synthesis of cultured fibroblasts. Our results show that term amniotic fluids are capable of inducing the synthesis of collagenase and other proteases in fibroblasts, as revealed by selective increases in collagenase activity and in immunoreactive collagenase. Nonterm amniotic fluids failed to do the same. This phenomenon is proposed as a model for studying the collagen degradation of fetal membranes during term gestation.

Evidence that human follicular fluid contains a chemoattractant for spermatozoa.

Considering that information from the literature clearly establishes that human follicular fluid enhances both motility and viability of spermatozoa and that this fluid contains substances with chemotactic activity for leukocytes, we looked to see if follicular fluid might exert any chemical attraction to male germinal cells. In an vitro model using 0.8% agarose plates, it was demonstrated that the number of cells migrating to wells containing follicular fluids from patients in a program for GIFT, classified as mature in accordance to oocyte morphology, was significantly higher than those migrating to control wells. Further studies are needed to confirm these results and to study the possible specificity of the signal.

[Impact of sperm capacitation on various populations of human spermatozoa]. / Impacto de la capacitación espermática en las diferentes poblaciones de espermatozoides humanos.

With the purpose of evaluating the impact of spermatic capacitation on different spermatozooa populations, 49 samples of semen, before and after in vitro spermatic capacitation with Ham F-10 medium, were studied; motility of cells was evaluated according to WHO criteria. There was diminution of percentage of immobile cells, 27.8 to 20.0, as well as increase in population of cells with more mobility, 28.6% to 39.1%. Both difference were statistically significant (p = less than 0.05 and p = less than 0.005, respectively). These data suggest that spermatic capacitacion activates "in cascade" all groups of gametes.