Autoantibody studies of female patients with reproductive failure.

The aim of this study was to investigate the prevalence and character of autoimmune derangements in women with reproductive failure. A total of 108 females (age range 17-43, mean 27.5 years), including 16 with primary menstrual cycle disturbances and polycystic ovaries (PCO), 20 with polycystic ovary syndrome (PCOS), 38 with endometriosis (E), and 34 with chronic anovulation, luteal phase insufficiency, subfertility or unexplained infertility (INF) were investigated. A control group of 392 women was formed from an unselected population sample (age range 17-43, mean 31.0 years). All sera were tested by indirect immunofluorescence method to assess common autoantibodies: nuclear (ANA), smooth muscle (SMA), parietal cell (PCA), thyroid microsomal (TMA), reticulin (ARA), mitochondrial (AMA) and liver/kidney microsomal autoantibodies (LKMA). Enzyme-linked immunosorbent assay was used to detect antibodies against beta2-glycoprotein I (anti-beta 2GPI) and carbonic anhydrase (anti-CA). Our results showed that 40.7% of patients' sera and 14.8% of control sera contained one or more common autoantibodies, ANA and SMA were most frequently detected (difference between two groups P<0.005). Anti-beta 2GPI were found in eight cases (7.4%), including two patients with INF but without other autoantibodies. Anti-CA were revealed in nine cases (8.3%) including patients' PCOS, E and INF. A comparison of patients' clinical data with antibody assay results did not reveal any significant associations. Our results indicate a high prevalence of autoimmune reactions in women with reproductive failure due to the most common causes PCO, PCOS and E as well as in unexplained infertility. This might reflect the propensity to develop autoimmune reactions in such patients, including pathogenic autoimmune reactions to specific target antigens.     

月经紊乱患者的细胞遗传学分析

目的 分析原发闭经、继发闭经及月经稀发患者的染色体核型,探讨性染色体异常对性腺发育的影响。方法 将176例患者分为两组,其中82例原发闭经组,94例继发闭经及月经稀发组。每例行外周血培养,制片及G显带,并行染色体核型分析。结果 176例患者发现性染色体异常38例,异常检出率为29.6％(38/176),其中原发闭经组33例,异常检出率为40.2％(33/82);继发闭经及月经稀发组检出性染色体异常5例,异常检出率为5.3％(5/94);两组异常检出率差异有显著性(P<0.05)。性染色体异常大体上分为三大类:含Y染色体(15例),X染色体数目异常(18例),X染色体结构异常(5例),嵌合体均以45,X系为主,共10例。结论 两条完整的染色体是女性性腺发育及正常卵巢功能所必须,性染色体异常是原发闭经的主要原因之一,常规细胞遗传学检查是必要的;继发闭经及月经稀发也不应忽视此项检查。     

The frequency of chromosomal abnormalities in patients with reproductive failure

Objective

To investigate the ratio of chromosomal abnormalities in recurrent fetal wastage.

Study design

We conducted a study of the cytogenetic data of 645 couples (1290 patients) with recurrent fetal wastage examined at the Department of Medical Biology and Genetics, Trabzon, Turkey. Couples who had first trimester miscarriages/abortion, preceded or followed by a second or third trimester fetal death/fetal abnormalities were recruited from Obstetrics and Gynecology Clinics for cytogenetics analysis.

Results

Chromosome abnormalities were found in 25 (3.86%) patients. The chromosomal abnormalities were structural (3.71%) and numerical (0.15%). Polymorphisms of heterochromatin blocks and inv(9) were shown in 115 (17.51%) patients.

Conclusions

Chromosome analyses are an important and necessary part of the etiological research in couples with recurrent fetal wastage.     

Immunologic studies in patients with premature ovarian failure

Tests for a range of autoantibodies, and counts of lymphocytes, B cells, T cells, and T cell subsets were performed in 45 Chinese patients with premature ovarian failure and 45 age-matched normal control subjects. Eight patients (18%) were positive for at least one autoantibody. Only one patient was positive for antiovarian antibody. Patients with autoantibodies had a significantly higher percentage of circulating B cells. The lymphocyte, T cell, CD4+, and CD8+ counts in patients with premature ovarian failure were significantly higher than those in the control group, but the CD4:CD8 ratio was significantly lower in women with premature ovarian failure. There was a significant negative correlation between plasma estradiol levels and CD8+ counts, and a significant positive correlation between plasma estradiol levels and CD4:CD8 ratios. The changes in lymphocytes and lymphocyte subpopulations in premature ovarian failure may be due to estrogen deficiency.     

Cytogenetic study of 201 subjects with altered reproductive fitness

The A. report a cytogenetic study performed on 201 subjects with a defective reproductive fitness. In total, they detected 37 chromosomal changes (18.4%): 7 in 48 subjects (24 couples) with sterility (14.5%), 11 in 96 with hypogonadism and/or criptorchidism (11.4%), 15 in 38 women with oligoamenorrhoea (39.4%) and 4 in 4 patients with Morris syndrome (100%). On the contrary, no chromosomal change was detected in 15 patients with pure gynecomastia. The A. discuss the significance of these chromosomal aberrations, and particularly: the mosaicism XO/XX, because this chromosome picture may be associated with fertility, as in our case; the isodicentric X, because the patient showed the clinical features of the Turner's syndrome; the inv(11), because the patient showed a progressive oligoamenorrhoea, leading us to retain that some chromosome changes, by determining a severe gametic selection, may cause infertility in these subjects. the t(13;14), detected in men with azoospermia, because these changes confirm the presence of a gametic selection, mostly in men. Therefore, according to other reports, the A. suggest that the cytogenetic investigations should be performed in all subjects with abnormal reproductive fitness, for a more accurate diagnostic iter.     

Cytogenetic studies in couples with repeated spontaneous abortions

Chromosome studies were carried out on both partners of 509 couples with a history of two or more spontaneous abortions. 1) Twenty-six individuals (2.6%) were carriers of a major chromosome abnormality. This incidence is at least six to seven times higher than that in the general adult population. 2) Of these, 10 were reciprocal translocations, 10 robertsonian translocations and 6 numerical aberrations of gonosomes. None of the carriers showed abnormal phenotypes. 3) Chromosome aberrations were more frequent in the women than in their husbands. There were 19 abnormalities in females and 7 in males. 4) The use of banding techniques in chromosome analysis improves the detection of balanced reciprocal translocations. 5) Prenatal diagnosis was performed in 5 subsequent pregnancies of 4 balanced translocation carriers. The fetal karyotypes were 2 normal and 3 balanced translocations. It would seem reasonable to recommend chromosome analysis for couples with repeated spontaneous abortions.     

Cytogenetic studies in infertility]

10 TO 15% of all married couples are undesired childless. New investigations show the tendency of a wider increase. Infertility is a possibility caused chromosomaly. In married couples carriers of balanced translocations causing repeated spontaneous abortions in a rate of ten times more than in the normal population. Every 50 married partner is a carrier. Chromosomal cause of infertility should be excluded before an operative therapy is planed and gives a base for genetic counseling.     

Cytogenetic studies in primary amenorrhea

Cytogenetic studies in 48 cases of primary amenorrhea are reported. The following abnormalities were analyzed: 3 cases of Turner's syndrome, 3 cases of Turner's mosaicism, 3 cases of feminizing testes, 2 cases of incomplete testicular feminization syndrome of Morris and Mahesh, 7 cases of Mullerian agenesis with absent vagina and functioning ovaries, 13 cases of gonadal dysgenesis with streak gonads, 1 case of pure gonadal dysgenesis, 2 cases of imperforate hymen with hematocolpos and hematometra, 1 case of tubovarian mass and 1 case of juvenile polycystic ovarian syndrome. It is suggested that management of an patient with primary amenorrhea whose sex karyotype includes an XY cell line should include gonadal excision because of increased risk of neoplasia.     

Cytogenetic studies in primary amenorrhea]

Cytogenetic analysis in 125 women with primary amenorrhea consisting of determinations of sex chromatin and karyotype, and in some cases of autoradiography were performed. On the basis of clinical, endocrinologic and cytogenetic criteria, the women were divided into ten clinical groups. In Turner's syndrome 45,X monosomie was observed only in 9 patients and in the remaining 12 cases varies types of mosaicism or of structural aberrations of the X chromosome. In pure gonadal dysgenesis, the patients exhibited 46,XY karyotype have the tendency to malign tumors of the gonads. In all cases with male pseudohermaphroditism the karyotypes 46,XY were observed. The remaining patients with primary amenorrhea exhibited 46,XX karyotype and belonged to the cases with Mayer-Rokitansky-Kustner syndrome, with adrenogenital syndrome, with hypoplasia of the ovaries, with primary amenorrhea of uterine or pituitary origin or at last with pubertas tarda.     

Cytogenetic studies of endometrial carcinoma

Chromosome counts have been obtained from 22 cases of carcinoma of the endometrium and 4 cases of atypical cystic hyperplasia. The direct squash technique after hypotonic pretreatment was used. A diploid population of 22.2 per cent was present in the cases of endometrial cardinoma which were analyzed. Eighty-eight per cent of these diploid metaphases exhibited a pseudodiploid karyotype. Ninety-three per cent of the total chromosome counts for endometrial carcinoma were in the range of 35 to 56 chromosomes per cell. The diploid population in the 4 cases of atypical hyperplasia was 66 per cent. Two cases were analyzed karyotypically. Normal female karyotypes appeared exclusively in one case. In the second case pseudodiploid karyotypes predominated, and rearrangement within the chromosome groups was evident in hypodiploid cells.     

Endometrial laminin subunit beta-3 expression associates with reproductive outcome in patients with repeated implantation failure

Purpose

Endometrial laminin subunit beta-3 (LAMB3) is a candidate gene whose expression distinguishes the endometrial window of receptivity (WOR) in human. This study aims to examine endometrial LAMB3 levels in patients with repeated implantation failure (RIF), in order to assess the ability of LAMB3 to predict pregnancy outcome.

Methods

Endometrial biopsies were taken during the WOR from 21 healthy volunteers in natural menstrual cycles and from 50 RIF patients in mock cycles prior to frozen embryo transfer (FET) cycles. Immunohistochemistry (IHC) staining of LAMB3 was performed, and the H-score was correlated with the pregnancy outcome in subsequent FETs.

Results

In healthy volunteers, endometrial LAMB3 was demonstrated to be highly expressed during the WOR with the staining exclusively in the cytoplasm of the epithelial cells. In a discovery set of RIF patients, the LAMB3 expression level was found to be significantly higher in those who conceived compared to those who did not in subsequent FETs. A receiving operator characteristic (ROC) analysis revealed an area under the curve (AUC) of 0.7818 (95% confidence interval 59.92–96.44%) with an H-score cutoff of 4.129 to differentiate cases with positive or negative pregnancy outcomes. This cutoff achieved an accuracy of 75% in pregnancy prediction in a following validation set of RIF patients, in which the pregnancy rate in subsequent FETs was three-fold higher when the mock cycle LAMB3 H-score was ≥ 4.129 compared to < 4.129.

Conclusions

IHC measurement of endometrial LAMB3 expression could be a promising prognostic method to predict pregnancy outcome for RIF patients undergoing FETs.

     

Cytogenetic analysis of 179 Iranian women with premature ovarian failure

Abstract

The importance of chromosomal abnormalities in etiology of premature ovarian failure (POF) is well known but in many cases, POF still remains idiopathic. We investigated the frequency and type of chromosomal aberrations in Iranian women diagnosed with idiopathic POF. Standard cytogenetic analysis was carried out in a total of 179 patients. Karyotype analysis of these patients revealed that 161 (89.95%) patients had normal female karyotype and 18 (10.05%) patients had abnormal karyotypes. The abnormal karyotypes included sex reverse sex determining region Y (SRY) negative (five Cases), X chromosome mosaicism (five cases), abnormal X chromosomes (three cases), abnormal autosomes (three cases) and X-autosome translocation (two cases). The overall prevalence of chromosomal abnormalities was 10.05% in this first large-scale report of chromosomal aberrations in Iranian women with POF. The results confirm previous observations and emphasis on the critical role of X chromosome abnormalities as one of the possible etiologies for POF.     

Mycoplasma and human reproductive failure. I. The occurrence of different Mycoplasmas in couples with reproductive failure

An epidemiological study of the occurrence of different Mycoplasmas in seminal fluid and in cervical secretions from patients with reproductive failure and in 2 control groups is described. All patients underwent a series of fertility tests before inclusion in the study and those with abnormal findings were excluded. Group A consisted of 36 couples in whom no cause of infertility could be found and group B of 19 couples in whom serum antibodies agglutinating donor sperm in different titers were detected in the women but not in the men. 2 control groups consisted of 40 women in group C attending a prenatal clinic and 23 men in group D married to women in the 3rd-9th month of pregnancy. Among the subjects, sperm specimens were obtained from the men and cervical swabs from the women on the 2nd or 3rd day of the menstrual period and cultured for Mycoplasmas. Cervical swabs taken after postcoital tests, and cervical and sperm specimens taken during the luteal phase were also cultured. Cervical and sperm specimens from the controls were also cultured. In group A, large colony-forming Mycoplamas were found in cervical secretions of 7 women and in seminal fluid of 2 men. T-Mycoplasmas were found in both spouses in 28 couples and in 3 women and 2 men with negative partners. In group B, large colony-forming Mycoplasmas were found in 2 women and 1 man whose partners were negative. T-Mycoplasmas were isolated from both spouses in 17 of the 19 couples and from the wife only in 2 couples. In group C, classical Mycoplasmas were found in 3 and T-Mycoplasmas in 9 of the 40 cases. In group D, no classical Mycoplasmas were discovered but T-Mycoplasmas were found in 6 of the 23 cases. The observed difference in frequency of T-Mycoplasmas between patient and control groups was highly significant statistically. T-Mycoplasmas were found growing from the spermatozoa but not from the supernate in 7 of 10 specimens obtained from men in group B. In 2 of the remaining 3 specimens, T-Mycoplasmas were found both in spermatozoa and in the supernate, while in the last specimen, no growth was found.     

Multiple autoantibodies associated with autoimmune reproductive failure

Purpose : Autoimmune factors are involved in some of the cases of reproductive failure. The aim of this paper is to discuss the association between autoantibodies and reproductive failure. Methods : Literature review of autoantibodies associated with reproductive failure. Results : Several autoantibodies were found in association with such clinical manifestations, mainly in patients having systemic lupus erythematosus or the antiphospholipid syndrome. These autoantibodies include classical antiphospholipid antibodies such as anticardiolipin, anti-2-glycoprotein-I, antiphosphatidylserine, and antiphosphatidylethanolamine. There are also some nonclassical antiphospholipid antibodies directed to prothrombin, thromboplastin, or mitochondrial antibodies of M5 type, which were also found in patients with reproductive failure. Moreover, animal models as well as some human studies support a role for other autoantibodies in these clinical manifestations including antithyroglobulin, antilaminin-1, anti-corpus luteum, antiprolactin, anti-poly(ADP-ribose), and lymphocytotoxic antibodies. Conclusions : Even though there is not enough data currently to support a firm association between some of these autoantibodies and reproductive failure, future studies are likely to help us determine and expand the number of autoantibodies screened in these patients.