Fertile Control Group Research Articles

The androgen receptor CAG repeat polymorphism and X-chromosome inactivation in Australian Caucasian women with infertility related to polycystic ovary syndrome.

The human androgen receptor (AR) gene contains a polymorphic trinucleotide (CAG) repeat sequence in exon 1. The number of CAG repeats may confer differential receptor activity, and specific ranges of variants have been correlated with androgen-sensitive disease processes. Polycystic ovary syndrome (PCOS) is a female condition characterized by androgen excess and infertility, many features of which are effected through the AR. We compared frequency distributions of CAG repeat alleles and their pattern of expression via X-inactivation analysis among 83 fertile women and 122 infertile women with PCOS, all of Australian Caucasian ethnicity. A population comparison with 831 predominantly fertile Australian women was also used. PCR-based assays were used to genotype each woman and assess allele inactivation patterns after digestion of DNA with methylation-sensitive HpaII. Infertile women with PCOS exhibited a greater frequency of CAG alleles or biallelic means greater than 22 repeats compared with both the fertile control group (P < 0.05) and the general population (P < 0.01). Preferential expression of longer CAG repeat alleles was also observed in PCOS and correlated with increased serum T. We conclude that the AR (CAG)n gene locus and/or its differential methylation patterns influence the disease process leading to PCOS.

Recurrent pregnancy loss: etiology of thrombophilia.

Congenital and acquired thrombophilia are associated with an increased risk of pregnancy-associated venous thrombosis and fetal loss. Two hundred eighty-nine patients with a history of recurrent spontaneous abortion were subjected to screening examinations for the etiology of these abortions. Endocrine abnormality (28.0%), uterine abnormality (10.4%), autoimmune diseases (1.4%), antiphospholipid antibody syndrome (4.5%), and balanced type chromosome translocation (4.2%) were found as underlying causes of recurrent abortions, and the remaining 55.0% of the 289 patients were classified as having an unexplained etiology. Congenital thrombophilia such as protein C (PC) deficiency, protein S (PS) deficiency, antithrombin deficiency, and factor V Leiden mutation was not frequently detected; only one patient had PS deficiency. A reduced factor XII activity was found at a frequency of 4.2%. The frequency of methylene tetrahydrofolate reductase gene C677T mutation in recurrent aborters (0.38) was the same as that found in a fertile control group. Although the prevalence of anti-beta2-glycoprotein I antibody (abeta2-GPI) syndrome was very low (1.7%), patients with a high titer of immunoglobulin G (IgG) class abeta2-GPI, despite anticoagulation therapy, experienced severe fetomaternal complications in subsequent pregnancies. The rate (13.8%) of positive tests for serum IgA class abeta2-GPI in patients with unexplained etiology was higher than that in the controls (0%) (P < .05). We conclude that congenital thrombophilia is rare in Japanese patients who had experienced consecutive spontaneous abortions.

Relationship between semen quality and the seminal plasma components carnitine, alpha-glucosidase, fructose, citrate and granulocyte elastase in infertile men compared with a normal population.

The seminal plasma components neutral alpha-glucosidase, carnitine, fructose, citrate, and polymorphonuclear granulocyte (PMN) elastase in 253 men were determined. The seminal plasma of 221 infertile men, a control group with proved fertility and 13 patients after vasectomy were investigated. The concentrations of free carnitine (212 versus 521 micromol/l, n = 219, P < 0.001), total carnitine (437 versus 743 micromol/l, n = 219, P < 0.001), and the activity of neutral alpha-glucosidase (15.1 versus 23.4 IU/l, n = 236, P < 0.05) were significantly reduced in the infertile patient group as compared to the fertile control group, the concentration of PMN elastase (102 versus 48 microg/l, n = 234, P < 0.05) being significantly increased in the infertile patients. In the patients after vasectomy the activity of neutral alpha-glucosidase was the only epididymal marker that was significantly reduced (4.3 versus 9. 8 IU/l, n = 35, P = 0.002) in comparison with the patients with testicular azoospermia. At a limit of 6 IU/l the sensitivity of the method was 92% and the specificity was 72%. Altogether, the epididymal markers were reduced in subfertile patients compared with the control group. For the differential diagnosis of azoospermia only the determination of the neutral alpha-glucosidase activity is useful.

Anti–β 2-glycoprotein I antibodies in women with recurrent spontaneous abortion, unexplained fetal death, and antiphospholipid syndrome

Objective: Studies in rheumatologic and hematologic populations suggest that anti–β 2-glycoprotein I antibodies are more specific for the clinical manifestations of antiphospholipid syndrome than anticardiolipin antibodies. However, the association between anti–β 2-glycoprotein I and pregnancy loss remains uncertain. We sought to determine whether anti–β 2-glycoprotein I is associated with the obstetric features of antiphospholipid syndrome. Study Design: Sera from 4 groups of women were studied: (1) 152 healthy fertile control subjects, (2) 141 subjects with unexplained recurrent spontaneous abortions, (3) 58 subjects with unexplained fetal deaths, and (4) 73 subjects with well-characterized antiphospholipid syndrome. Serum anticardiolipin and anti–β 2-glycoprotein I levels were determined by enzyme-linked immunoassay. Results: Patients with antiphospholipid syndrome had significantly higher levels of immunoglobulin G and immunoglobulin M anticardiolipin and anti–β 2-glycoprotein I than the other 3 groups ( P < .0001). However, women in the recurrent spontaneous abortion, fetal death, and fertile control groups had similar levels of each antibody. Similarly, there were no differences in the proportion of women with positive test results for each autoantibody in these 3 groups. Linear regression analysis showed significant correlation between anticardiolipin immunoglobulin G and β 2-glycoprotein I immunoglobulin G ( R 2 = 0.544786, P = .0001) and anticardiolipin immunoglobulin M and β 2-glycoprotein I immunoglobulin M ( R 2 = 0.525048, P = .0001). Conclusion: Both anticardiolipin and anti–β 2-glycoprotein I are associated with antiphospholipid syndrome. However, testing for anti–β 2-glycoprotein I does not identify additional patients with either recurrent spontaneous abortions or unexplained fetal deaths who initially have negative test responses for anticardiolipin. This is likely because of the strong correlation between the 2 autoantibodies. Our data do not support routine testing for anti–β 2-glycoprotein I in addition to testing for antiphospholipid antibodies in women with recurrent pregnancy loss and unexplained fetal death. (Am J Obstet Gynecol 1999;181:642-8.)

Differences in the DNA-stainability of spermatozoa from fertile and suspected infertile men.

The aim of this work was to determine whether it is possible to distinguish between fertile (control group, already fathers) and infertile men (suspected infertility), by comparing the fluorescence intensity of the sperm-DNA after incubation with appropriate dyes. First we examined two different DNA-specific dyes (DAPI and YOYO-1) using bull spermatozoa. Based on good results in immunohistochemical applications, YOYO-1 was chosen for further work. The fluorescence-intensity of 200 single, morphologically normal spermatozoa in each semen sample were measured in a cytophotometer, means + SD determined and histograms delineated. Of 20 samples from the control group, 17 had markedly higher fluorescence-intensity than did 7/15 of the suspected infertile men. It is concluded that the DNA of the latter seven samples was less accessible to the dye than was the DNA of the control group. There are cases of infertility known in which there is loss of one or more of the DNA-binding proteins, which in spermatozoa are mainly (85%) protamines. The relationship between the stainability of the sperm-DNA and the packaging with DNA-binding proteins is discussed. Two of the histograms showed abnormalities in the distribution of the fluorescence-intensities, one sample was extremely fragile and most of the sperm lysed during the staining-procedure. Five samples showed normal histograms in comparison with the control group.

EFFECTS OF SUBINGUINAL VARICOCELE LIGATION ON SPERM CONCENTRATION, MOTILITY AND KRUGER MORPHOLOGY

Purpose We examined the effects of varicocelectomy on semen parameters in 30 subfertile men, with emphasis on potential changes in sperm count, motility and morphology as measured by Kruger's strict morphologic criteria. Materials and Methods A total of 30 patients underwent subinguinal varicocelectomy (25 bilateral and 5 unilateral). Preoperative and postoperative sperm density, motility and morphology were analyzed. Preoperative follicle-stimulating hormone, luteinizing hormone and testosterone levels were measured and compared to those of fertile volunteers enrolled in our sperm donation program. Pregnancy rates after varicocelectomy were also examined. The Wilcoxon signed rank test was used to measure levels of statistical significance in all analyses. Results We found that sperm density and motility improved significantly (p <0.05) without concomitant changes in strict morphology (p >0.05) only in men with clinical bilateral varicoceles. No differences were observed in values among testosterone, follicle-stimulating hormone and luteinizing hormone levels of the fertile control group and preoperative varicocele patients. Of 30 patients 12 (40%) had successful, full-term pregnancies, including 6 via natural cycle intercourse, 5 (43%) by in vitro fertilization embryo transfer and 1 by intracytoplasmic sperm injection. Conclusions Although sperm morphology as measured by strict morphologic criteria does not improve after varicocelectomy, there were highly significant changes in motility and concentration. Hormonal differences are not likely to have a role in or be reflective of pathophysiology of varicocele induced male infertility. The recent observation that sperm motility may be an independent or additive predictive factor for fertilization and pregnancy supports the need for continued varicocele repair independent of the lack of varicocelectomy effect on Kruger morphology.

Idiopathic habitual abortion: experiences with active immunotherapy

The effect of lymphocyte transfusions on the stabilisation and successful outcome of pregnancy was investigated in 30 women with recurrent abortions presumably caused by immunologic factors. Lymphocyte concentrates were obtained from stored whole blood of the husbands and transferred by intravenous injection. Exclusion criteria for this study were the presence of antipaternal lymphocytotoxic antibodies in the patient's serum (cross match) or an advanced pregnancy (> 12 weeks). After transfusion of paternal lymphocytes 19 (63%) women had uncomplicated and successful pregnancies whereas 11 (37%) suffered again from early pregnancy loss. Two out of these 11 patients aborted twice. The probability of a successful treatment decreased as soon as lymphocytotoxic antibodies appeared in the patient's serum (p < or = 0.021). Women with a lower number of abortions before treatment carried their pregnancies to full term more frequently (p < or = 0.03). The number of transfusions, the period between start of treatment and conception as well as the distribution of age within the two groups had no significant effect on the outcome of therapy. A lack of lymphocytotoxic antibodies or their late appearance during treatment give a positive prognosis for the progress of pregnancy. Compared with a fertile control group, couples with habitual abortions showed no significantly increased HLA sharing. Immunisation with paternal lymphocytes in cases of presumably habitual abortions due to immunologic factors appears as an effective therapy with only few side effects. Large, randomised, double-blind, and placebo-controlled trials are needed before drawing conclusions.

Validation of an embryotoxicity assay.

Culture of mouse blastocysts has served as a tool for identifying various embryotoxic factors in human serum. While inactivated, sera from recurrently aborting women inhibit mouse blastocyst development in vitro. Variation in results from individual serum samples has limited the usefulness of this assay in establishing a new classification of idiopathic recurrent spontaneous abortion (RSA). Two-cell embryos were collected from superovulated mated CB6F1/J mice and cultured in Ham's F-10 media supplemented with 10% fetal bovine serum (FBS) or tested human serum at 37 degrees C with 5% CO2 and high humidity. Each sample was assayed in triplicate using three mice with at least five embryos from the same mouse per dish. Development was evaluated at 72 h and the frequency of atretic embryos was recorded. Intrasample (interassay) variation yielded a coefficient of variation of 9%. When repeated, samples from a given individual were evaluated and the coefficient of variation was 8.7%. Interoperator variability was 4% interassay and 2% intrassay. Atresia of embryos was 23% when incubated with FBS (N = 122), 21% in FC (N = 122), and in the sera of patients with RSA 34.6% (N = 95). Results of percentage of atresia from the fertile control group had a nonparametric distribution. Using 2.2 multiples of the median to determine the 95% confidence interval, a threshold at 44.0% of atresia was established. The critical step in maintaining low variability in this bioassay is to control mouse variability by averaging the percentage atresia from different mice as embryo donors for each tested serum. A subgroup of 24% (23/95) RSA patients who displayed embryotoxic activity was identified with a specificity of 95% and positive predictive value of 83%, P = 0.001.

Increases in chromosome aberrations and in abnormal sperm morphology in rubber factory workers

Subjects working at a rubber plant in a chemicals warehouse or in calandering and bambury units were analyzed for both sperm parameters and structural chromosome aberrations in peripheral blood lymphocytes. Sperm analysis war performed in a group of 24 workers for comparison with fertile (n = 24) and infertile (n = 24) control groups. The statistical analyses of semen volume, vitality and sperm count did not show significant differences between exposed and fertile groups but significant differences were found from the infertile group. A significantly lower proportion of normal sperm head shapes was found in exposed subjects when compared to the fertile group (40.1 vs. 57.8). Seven exposed workers were re-analyzed 1 year and their sperm parameters did not change. The cytogenetic analysis showed a significant increase (3.90%) in the percentage of cells with aberrations in bambury workers (n = 11). However, no differences were found between calandering workers (n = 8) and control subjects (n = 10). Workplace air samples taken on the day of tissue sampling did not show any increase above the Cuban maximal allowed concentration for benzo[ a]pyrene or toluene.

Effect of obesity and fertility status on sex steroid levels in men

Endocrine studies were performed on fertile and infertile obese men and compared with fertile and infertile nonobese men in order to determine the independent and codependent effects of obesity and fertility status on the male hypothalamic-pituitary gonadal axis. The obese infertile group exhibited significant endocrinologic changes as compared with fertile nonobese control group which was not observed in any of the other three groups. Serum testosterone was significantly lower. The testosterone/estradiol ratio was significantly lower despite a lack of significant change in serum estradiol levels. Serum steroid hormone binding globulin (SHBG) was significantly lower which correlated with elevated bioavailability of both testosterone and estradiol in the obese infertile group. Serum luteinizing hormone levels were no different, suggesting that free testosterone levels were unchanged. Obese infertile men exhibit endocrinologic changes that are not observed in men with either obesity or infertility alone. Reduction of serum SHBG, total testosterone, and testosterone/estradiol ratio appear to be a marker of infertility among obese men.

Incidence of sperm with two fluorescent bodies in men with impaired fertility

To determine the two fluorescent body (2FLB) frequency in males with impaired fertility. Males of couples were evaluated along with their female partners for reproductive function. They were classified as having the potential for fertility (control group) or as infertile (test group). This was based on the evaluation of both members of the couple. Semen parameters were determined and analyzed for differences among diagnostic groups. The setting was clinical secondary care in both private and institutional practice. The study population was composed of males of couples who were being evaluated for infertility. Classification as to reproductive potential and categories of abnormality was based on measurement of semen parameters and clinical judgment. Semen specimens were obtained and analyzed from 78 control and 93 test subjects. Semen parameters including volume, sperm count, sperm motility, percent morphologically abnormal sperm, and percent of quinacrine-stained sperm containing zero, one, or two FLBs were determined. The test group was found to have a 2FLB frequency that was significantly different from that of the control group. The data were then analyzed for significant differences in 2FLB frequencies in subpopulations. When subjects from all diagnostic subgroups that included varicocele were compared with the controls, the 2FLB frequency was significantly elevated (median percent of 2FLB was 1.2 versus 0.7). The 2FLB frequency in the remaining diagnostic groups was not elevated. This is the first report of an elevated 2FLB frequency associated with a biological abnormality. The cause of the elevated 2FLB frequency is not known. However, other anomalies that have been identified in varicocele patients have been associated with elevated intrascrotal temperatures. There are data available that associates elevated temperature with aneuploidy in mouse oocytes.

Time-Related Decline in Sperm Motility Patterns in Men with Cytotoxic Antibodies

Sperm motility patterns in semen from 10 fertile nonautoimmune men (fertile control group) and 33 infertile men with various titers of cytotoxic sperm antibodies in their seminal plasma (group 1: titers less than or equal to 16, n = 6; group 2: titers 64 to 512, n = 12; group 3: titers greater than or equal to 1024, n = 15) were evaluated every 2 hours for 12 hours and finally at 24 hours. A significant decline in sperm swimming speed and linearity was observed at 6 hours in semen from 27 infertile men with sperm antibodies. Beginning at 8 hours, semen from sperm antibody-positive men in group 2 showed a significant decline in percentage motile sperm (p less than 0.001) compared to the fertile controls. The percentage motility in semen of donors in groups 1 and 3 was significantly lower than that in semen of fertile donors at 10 hours (p less than 0.05), 12 hours (p less than 0.01), and 24 hours (p less than 0.001). The mean velocity in groups 2 and 3 was significantly less than that in fertile controls at 10 and 12 hours (p less than 0.05). The linearity of sperm motility started to decline 4 hours after semen samples were obtained from sperm antibody-positive groups 2 and 3 in contrast to sperm antibody-negative fertile or infertile groups (p less than 0.05). It is concluded that the presence of cytotoxic sperm antibodies in the seminal plasma adversely affects sperm linearity as early as 4 hours after semen collection.(ABSTRACT TRUNCATED AT 250 WORDS)

Morphology of live seminal and postcoital cervical spermatozoa and its bearing on human fertility.

Various seminal variables were studied in two groups of infertile couples and in one fertile control group. Sperm morphology was emphasized. Patients with normal postcoital tests and sperm counts exceeding 5 mill/ml were selected and followed prospectively. The two clinical groups could then be identified: a. couples who were persistently infertile during the 3 years of observation and whose infertility was not explained by the clinical investigation, and; b. couples where conception was achieved during the period of observation, either spontaneously or after treatment of the woman. In the latter group all the variables of semen analysis were identical with those of the control group. Differences between the two groups of infertile patients were noted only with respect to the morphology of live spermatozoa of the semen sample and of postcoital spermatozoa within the cervical secretion. It is concluded that these two variables of sperm evaluation are of prognostic significance with regard to future fertility.

Peritoneal fluid prostanoids in unexplained infertility

Elevated peritoneal fluid thromboxane B2 and 6-keto-prostaglandin F1α have been reported in patients with biopsy-proved endometriosis. In this paper these peritoneal fluid prostanoids were measured in patients with unexplained infertility and a fertile control group matched for age and day of the menstrual cycle. A subgroup of the women with unexplained infertility demonstrated a marked elevation of both prostanoids (p < 0.01). These elevations may serve to identify women in whom undetected endometriosis or some other peritoneal irritant is associated with infertility.

An analysis of sperm function in cases of unexplained infertility: conventional criteria, movement characteristics, and fertilizing capacity

A detailed analysis of semen quality was carried out in 85 couples with unexplained infertility by the use of conventional criteria of semen analysis, time-exposure photomicrography, and the zona-free hamster egg penetration test. According to the latter, 34.1% of the male partners exhibited evidence of defective sperm function, although only 4% of these patients were devoid of any demonstrable fertilizing capacity. Of the conventional parameters of semen analysis examined, the most revealing was the morphologic character of the sperm, which was significantly poorer (P less than 0.001) in the group with unexplained infertility than in the normal fertile control group and also showed a significant relationship (P less than 0.001) with the presence of subnormal fertilizing capacity in the hamster egg assay. A majority of movement characteristics measured by time-exposure photomicrography were significantly depressed in the group with unexplained infertility, compared with the normal fertile control group. In addition, certain of the movement characteristics investigated were significantly related to the fertilizing capacity of the spermatozoa; and, in this respect, a progressive swimming speed (greater than 25 micrometers/sec), a straight swimming mode of progression, and a small amplitude of lateral head displacement (Ah) all appeared to be important qualities.

Searching for the infertile man with hyperprolactinemia

The purpose of this study was to determine whether routine prolactin measurement was of use when investigating men with infertility. Prolactin levels were slightly higher in men with primary infertility, compared with men with secondary infertility and a fertile control group. Although this slight increase was statistically significant, most readings were within the laboratory normal range in all groups, and we did not find any clinically significant cases of hyperprolactinemia. We conclude that routine prolactin estimation is not justified unless there are other indications, e.g., impotence, dialysis, or a history of previous pituitary disease.

Are infertile men and women neurotic?

Abstract Forty-seven infertile married women, and forty-seven married women in each of two fertile control groups, completed the Neuroticism Scale Questionnaire. Forty-five husbands of the infertile women and a total of eighty-two fertile husbands also completed the Questionnaire. On the basis of previous studies that have mainly used clinical interviews and projective tests, the hypothesis was tested that infertile Ss have higher mean Neuroticism than fertile Ss. It was found that neither infertile wives nor husbands differed from the fertile control groups, contrary to the hypothesis.