Cytogenetic investigations in couples with repeated miscarriages and malformed children: report of a novel insertion

Recurrent miscarriage and the birth of a malformed child continue to be challenging problems for the patient and clinician. Often, cytogenetic studies have an important role in the evaluation of couples with a poor obstetric history. The present study deals with analysis of chromosome abnormalities in 2150 couples from India, and is the largest group ever reported in the literature. The observed incidence of chromosomal rearrangements in these couples was 3.5%. Apart from reciprocal, Robertsonian, inversions, a unique case of chromosome insertion was identified, which is perhaps only the second report in literature. Minor chromosome variants such as inv(9),inv(Y),9qh+, D/G variants were observed in 108 subjects. Cytogenetic studies should be performed for all couples with repeated miscarriages and bad obstetric history, and in cases of detected chromosomal aberration, the patient should be counselled individually according to the type of anomaly.     

Six kinds of anti-phospholipid antibodies (aPLs) in ovulatory mucus and seminal plasma from couples with repeated miscarriages

OBJECTIVE: Determination of aPLs in ovulatory mucus and in seminal plasma in 18 infertile couples (women, median age 33.2, range 28-39 years; men, median age 36.4, range 29-46 years) with repeated unexplained miscarriages (four to nine spontaneous abortions). Women with high levels of aPLs in sera were excluded from the study. METHODS: aPL screening included cardiolipin (Sevac Prague), L-alpha-phosphatidic acid, L-alpha-phosphatidylethanolamine, L-alpha-phosphatidyl-DL-glycerol, L-alpha-phosphatidylinositol, and L-alpha-phosphatidyl/serine (Sigma, USA). These were determined in isotypes IgG and IgA by an ELISA method. Statistical analysis of aPL results was made by cut-off levels of aPLs for both Ig isotypes by using 3 SD or 95th percentile calculated by STATGRAPHICS. Control group was created from four fertile healthy couples. RESULTS: Levels of aPLs in ovulatory cervical mucus were in the majority very low, except for IgA-aPLs-inositol. Very high interindividual differences in aPLs were found in seminal plasma. IgA-aPL-ethanolamine and -DL-glycerol, and IgG-aPL-L-serine and -DL-glycerol predominated in seminal plasma. Extremely high IgA levels of aPLs in all above-mentioned antigens, except cardiolipin, were found in one man of an infertile couple. CONCLUSION: The passage of abnormal aPLs levels from seminal plasma into the site of conception during often repeated, unprotected intercourses is supported by the findings.     

Chromosome studies in couples with repeated abortions

In a nonselected group of 26 women with 3 or more abortions and in 17 of the respective husbands, chromosomes from lymphocyte cultures were investigated. 6 persons were found to have abnormal chromosomes; in 3 women variations in the length of autosomes were found (46,XX,Gp+; 46,XX ,Gp-; 46,XX,Cq+). 1 woman showed a trisomy C mosaicism in 4% of the analyzed cells. A second one had in 4% of the mitoses a translocation mosaic with a B/D-translocation(46,XX/46,XX,t(Bq+; Dq-)). Another woman with normal karyotype showed in 3 different analyses an increased rate of secondary chromosome aberrations with a high percentage of exchanges.     

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.     

The role of parental translocations in cases of repeated miscarriages

In families in which spontaneous miscarriages are very frequent, a systematic search for parental translocations must be made. An examination of the karyotype appears to be justified in patients with three or more miscarriages (probabilily of detecting a translocation equal to or greater than 1 percent). The results of cytogenetic analysis will indicate in many cases the prognosis for subsequent pregnancies and will to a large extent determine the procedure to be followed.     

Chromosome study of couples with repeated spontaneous abortions

The incidence of chromosomal abnormalities was evaluated in couples with a history of repeated abortions. The study included 14 couples with a history of 2 or more spontaneous abortions. Chromosome analysis was carried out on both marital partners from peripheral lymphocyte cultures. In every case, 30-62 metaphases were counted, and at last 20 karyotyped. Among the 14 couples, chromosome anomaly was found in 1 of the partners in 1 case. The patient, a 29-year-old female, was admitted to the hospital in the 36th week of her 10th pregnancy. In addition to an induced abortion and a premature delivery, her history included 7 spontaneous abortions at the 1st or 2nd month. Chromosome analysis of her peripheral blood revealed neither numerical nor structural aberations. In the 38th week, the patient gave birth ato a 3990 gm healthy boy whose karyotype of the cord blood also proved to be normal. The karyotype of the patient's husband revealed 45 chromosomes in 32 observed cells, and a chromosome was missing from each of the D and G groups. It was not possible to include in any of the groups a medium-sized, subterminal chromosome, diagnosed as D/G translocation. The husband proved to be a balanced D/G translocation carrier. When the pedigree was prepared, it was found that the younger sister of the husband had given birth to a child 1 month before term. Both the sister and her child were studied. The sister proved to be a balanced D/G translocation carrier. The child had translocational Down's syndrome: the D/G translocation occured in an unbalanced form. A table lists the results of chromosome investigations of cases of repeated abortions. Of 1066 married partners, chromosome aberations or variations responsible for multiple abortions were found in 70 (6.5%). Of these, 26 proved to be translocations. This incidence, 2-4%, is higher than the .4% in the general population. The results add credence to the argument that chromosome aberation may play a role in the causal background of recurring abortions. It is possible that with special staining procedures some aberations can be demonstrated in cases hitherto considered normal. In cases of repeated abortion, cytogenetic examination of both partners is routinely justified. In cases of chromosome aberations valuable data can be gained to serve as a basis for genetic counseling. When examining women with recurrent abortions, it should be possible to select patients to whom the possibility of intrauterine cytogenetic diagnosis can be offered.     

Chromosomal analysis in Japanese couples with repeated spontaneous abortions

Balanced chromosome rearrangements have been found at an increased frequency in couples with pregnancy wastage, especially recurrent spontaneous abortions, compared with the general population. In the present study, chromosomal analysis of peripheral blood cells, as one of the routine examinations of patients with repeated reproductive wastage, was performed on both partners of 639 Japanese couples. Among the 639 couples, 32 major chromosomal anomalies (5.0%) and 23 minor chromosomal variants (3.6%) were found. Both partners of one couple had an abnormal karyotype. The 32 major anomalies consisted of 19 reciprocal translocations, 9 Robertsonian translocations, one large inversion, two triple-X females, and one Turner mosaicism. The 23 minor variants included 15 cases of pericentric inversion of chromosome 9. The total number of pregnancies in the 54 couples with chromosomal anomalies was 181, but they resulted in only 18 normal liveborn neonates, indicating a 90.1% abortion rate. The present statistical study indicates that major chromosomal anomalies seem to be involved in repeated reproductive wastage.     

Mutation analysis of three genes in patients with maturation arrest of spermatogenesis and couples with recurrent miscarriages

The primary aim of this study was to gain more insight into maturation arrest of spermatogenesis (MA) and its relationship with mutations in genes essential for meiosis. The study also investigated the possibility that mutations in human meiosis genes cause a milder phenotype and that, in such cases, meiosis could potentially be completed with the production of mature germ cells having an abnormal chromosomal constitution causing miscarriage. Among 40 patients with MA, five changes were observed that also predicted alterations at the amino acid level. However, since these changes were also present in men with normozoospermia in equal frequencies, it was assumed that these changes are single nucleotide polymorphisms. Among 46 patients with recurrent miscarriages, two additional changes were detected predicting an alteration at the amino acid level. One change was detected in controls. However, the second heterozygous change, detected in a conserved functional domain of the SYCP3 gene, was absent in >200 controls. These preliminary results stress the need to further investigate the relationship between abnormalities in meiosis genes and the formation of gametes with abnormal chromosomal constitution. More research is also necessary to determine the impact and frequency of such changes before implementing mutation screening in genetic counselling.     

Cytogenetic abnormalities in 1162 couples with recurrent miscarriages in Southern region of India: report and review

Purpose  

The purpose of the present study was to investigate the contribution of chromosomal anomalies and the frequency of a particular type of aberration in couples with recurrent miscarriages.     

Treatment of severe rhesus isoimmunization by repeated transfusions in utero]

Repeated transfusions in utero by umbilical cord puncture enable the treatment of severe forms of rhesus incompatibility. The two cases reported, including one involving seven transfusions between twenty and thirty seven weeks since the LMP, form the basis of a review of the literature defining the advantages and results of this technique.     

Thyroid autoimmunity in patients with recurrent spontaneous miscarriages.

Recurrent spontaneous miscarriage (RSM) is a multifactorial problem. Auto- and alloimmune parameters have been implicated. Antithyroid antibodies (ATA) were tested in a group of women with RSM. The presence of antipaternal antibodies (APCA) was evaluated as an index of alloimmune contribution. Thirty euthyroid women with RSM (three or more consecutive miscarriages) aged 25-37 years were compared with 15 matched controls. Thyroid peroxidase (TPO) and thyroglobulin antibodies were tested with a chemiluminescence immunoassay and APCA were tested with a cross-match reaction. Results were compared using the chi-squared test. There was a higher frequency of ATA in women with RSM compared to controls (37% versus 13%, p < 0.05). Twenty of the women (67%) with RSM were tested negative for APCA, indicating an alloimmune contribution to their infertility. In this subgroup of women, the frequency of ATA continued to be higher than controls (40% versus 13%, p < 0.05). In conclusion, women with RSM, independent of APCA status, have a higher frequency of ATA. This may represent an additional marker for impaired regulation of the maternal immune system.     

Embryoscopy and karyotype findings of repeated miscarriages in recurrent pregnancy loss and spontaneous pregnancy loss

Purpose

The aim of the study was to assess cytogenetic and embryoscopic characteristics in subsequent miscarriages of spontaneous pregnancy losses (SPL) and recurrent pregnancy losses (RPL).

Methods

A retrospective cohort of 75 women was affected by repeated pregnancy loss. Of those, 34 had SPL, 24 primary RPL, and 17 secondary RPL. Ploidy status and morphology was analyzed by transcervical embryoscopic examination of the embryo and cytogenetic analysis of the chorionic villi in subsequent miscarriages.

Results

Similar rates of recurrent ploidy status were observed between first and second miscarriage in SPL and RPL (82.4% recurrent ploidy status in SPL, p?>?0.999; 73% recurrent ploidy status in RPL, p?=?0.227). No difference was found regarding recurrent abnormal morphology between SPL and RPL (p?=?0.092). However, secondary RPL resulted significantly more often in recurrent abnormal morphology compared to primary RPL (p?=?0.004).

Conclusions

High rates of recurrent normal/abnormal karyotypes were observed in all groups with a majority of embryos presenting with recurrent abnormal morphology. Secondary RPL presented significantly more often with recurrent abnormal morphology compared to primary RPL. These findings offer prognostic information for the affected patient and might impact treatment choice.

     

Detection of balanced chromosome rearrangements in 445 couples with repeated abortion and cytogenetic prenatal testing in carriers

Cytogenetic studies were performed in 445 couples presenting because of repetitive abortion. The authors detected a balanced translocation in 19 (4.2%) of the couples, pericentric inversions in 8 (1.8%), and polymorphisms in 52 (11.4%). The results were compared with those obtained in a series of 600 consecutive normal liveborns. Significantly higher frequencies for translocations and polymorphisms were present in couples with repetitive abortion. No sex predominance in translocation carriers was observed, and reciprocal translocations were more common (16 of 19) than the robertsonian type (3 of 19). Contrary to other reports, all of the translocations and inversions were detected among couples without previous abnormal offspring. Cytogenetic prenatal testing in 17 pregnancies from a carrier parent showed that none of the fetuses had the unbalanced karyotype, 13 carried the balanced rearrangement, and 4 had a normal karyotype. Although a risk profile can be obtained for these couples, prenatal testing must be offered to avoid anxiety and unjustified pregnancy interruptions.     

Anti-annexin as a marker in patients with recurrent miscarriages

ObjectiveTo investigate the possible role of anti-annexin A5 antibodies in inducing recurrent miscarriage.DesignCross-sectional study.SettingBenha University Hospital.Participants and interventionsPregnant women during the first trimester of pregnancy with a history of three or more consecutive pregnancy losses were compared to Multigravidae women during the first trimester of pregnancy with no history of pregnancy losses for group II. Women were screened for anti-annexin A5 antibodies.ResultsThere were no statistically significant correlations between anti-annexin A5 antibodies in both groups (P>0.05).ConclusionThe present study did not confirm that anti-annexin A5 antibodies have significance in predicting RM when measured in healthy women.     

Cytogenetic investigation in 413 couples with spontaneous abortions.

413 couples with one or more spontaneous miscarriages were karyotyped. Observed chromosomal rearrangements were classified as major rearrangements, i.e. Robertsonian and reciprocal translocations, pericentric and paracentric inversions, supernumerary small metacentrics, and minor rearrangements, i.e. pericentric inversions of chromosome 9, and constitutional fragility of particular chromosome sites. 2.30% of the individuals were carriers of a major rearrangement, which represents a ten-fold increase when compared to the general population. The contribution of each type of rearrangement is unequal, the most important being pericentric inversions (36 times more frequent than in the general population). Contrary to data from the literature, the probability of finding a rearrangement does not seem to increase with the number of miscarriages.     

White blood cell immunization and anticardiolipin antibody levels in women with recurrent miscarriages.

To determine whether WBC immunization stimulates production of anticardiolipin antibodies, anticardiolipin antibodies were measured before and 6 weeks after WBC immunization. Twenty-four non-pregnant women, who had had recurrent miscarriages for which a definitive cause could not be determined, were immunized with their partner's WBC. No significant differences in levels of anticardiolipin antibodies were detected between paired samples of sera obtained before and 6 weeks after WBC immunization. White cell immunization in nonpregnant women did not stimulate production of anticardiolipin antibodies.