Muhterem Bahçe

Chromosome abnormalities in 1255 cleavage-stage human embryos

The relationship was examined between chromosome abnormalities in cleavage stage human embryos and maternal age, embryo morphology and development rate. Embryos that were classified as suboptimal for transfer from patients undergoing IVF treatment were disaggregated, and all or most of their cells were fixed for analysis by fluorescence in-situ hybridization. Chromosomes X, Y, 13, 18 and 21, and in some instances 16 were examined. A total of 731 non-viable embryos was analysed. An increase in chromosome abnormalities with decreasing embryo competence and increasing maternal age was shown. Compared with an earlier study, the major difference was that polyploidy (P<00.01) and aneuploidy were previously more common. After pooling results, it was found that aneuploidy increased with maternal age, from 3.1% in embryos from 20-34 years old patients to 17% in patients 40 years or older. Also, aneuploidy occurred more frequently in embryos with good morphology and development rate than in embryos developing poorly. In contrast, dysmorphic and slowly developing or arrested embryos had significantly more polyploidy and mosaicism than normally developing embryos. Clear associations between maternal age and aneuploidy, and between cleavage anomalies and mosaicism have been established in non-viable embryos. Arrested embryos were mostly polyploid. Moreover, polyploidy was found more frequently in embryos analysed on day 4, suggesting that developmentally compromised embryos became arrested in extended culture. A slightly higher aneuploidy rate in the earlier study may be attributed to differences in hormonal stimulation, which also resulted in different numbers of oocytes recruited and matured.

Case report: chromatid exchange and predivision of chromatids as other sources of abnormal oocytes detected by preimplantation genetic diagnosis of translocations

Preimplantation genetic diagnosis of translocations can be performed on first polar bodies (PB) at metaphase stage using FISH with whole‐chromosome painting DNA probes. Here we report the use of this method in a couple in which the female was a carrier of a balanced translocation 46, XX, t(11;16)(q21;q22). This case was unusual in that two polar bodies showed recombination events between the homologue chromosomes of 11 and 16 pairs, resulting in M‐II oocytes with monovalent chromosomes having a normal and a derivative chromatid. For this type of case, PGD analysis on polar bodies cannot give a useful result, because, at the second meiotic division, either of these chromatids could remain in the oocyte, resulting in a normal, balanced or unbalanced embryo. PGD analysis on blastomeres can provide a solution. 11 previous cases of PGD of translocations performed by metaphase PB analysis are reviewed. Copyright

Thrombophilia-associated gene mutations in women with pregnancies complicated by fetal neural tube defects

Neural tube defects (NTDs) are one of the most common fetal defects. The causes of these defects are considered to have multiple factors involving nutritional deficiencies, genetic predisposition, and environmental factors such as drug exposure [1]. The aim of our study was to investigate the frequencies of thrombophilia-associated gene factor V Leiden mutations and prothrombin gene G20210A mutations, as well as methylenetetrahydrofolate reductase MTHFR C677T mutation which has a substantial role in the folate mechanism of women experiencing pregnancies with NTDs. This study consisted of 2 groups: group 1 consisted of 29 women whose previous pregnancies were complicated with NTDs, and group 2 was the control group consisting of 35 women who experienced uncomplicated pregnancies. The ages of patients in the control group matched with those in the NTD group. Case characteristics are displayed in Table 1. The frequency of theMTHFRC677Tmutationwas found to be significantly higher in women who experienced pregnancies withNTDs,while occurrence of factor V Leiden andprothrombin gene G20210A mutations were not significantly different between the study and control groups. The frequency of both homozygous and heterozygous MTHFR C677T mutation was 69.0% inwomenwho experienced pregnancies with NTDs (42.9% in the control group). Thepresence ofMTHFRC677Thad an odds ratio of 2.96 (95.0%CI, 1.05–8.32) for the development of NTDs. The frequencies of factor V Leiden, prothrombin geneG20210A, and MTHFR C677T mutations and allele frequencies with statistical comparisons are shown in Table 1. A study by Akar et al. [2] showed no relationship between MTHFR C677T mutation and spina bifida. However, they suggested that coexpression of MTHFR A1298C and MTHFR C677T mutations increased the risk of spina bifida. A similar association for NTD cases was reported in another study [3]. ⁎ Corresponding author. Department of Obstetrics and Gynecology, Gulhane Military Medical Academy, Etlik, 06010 Ankara, Turkey. Tel.: +90 312 3045818; fax: +90 312 3045800. E-mail address: temelceyhan@yahoo.com (S.T. Ceyhan).

Application of the ‘Apt test’ in Prenatal Diagnosis to evaluate the Fetal Origin of Blood obtained by Cordocentesis: results of 30 pregnancies

This study aimed to set up a practical lab‐side approach to discriminate fetal from maternal blood in samples obtained by cordocentesis. To determine the fetal origin of the blood, a modified Apt test was applied to 30 cases of prenatal diagnosis. A change of colour of the fetal and adult blood during the procedure was the hallmark to assess fetal origin. At the end of 60 s of the test, fetal blood yielded a pink colour whereas adult blood was dark green‐brown. The test was repeated in mixtures of fetal and adult blood. The results suggest that the modified Apt test is a practical, quick, inexpensive, and efficient test to determine the origin of blood samples obtained by cordocentesis. However, it should be kept in mind that samples containing a mixture of both fetal and adult blood could also yield a fetal blood reaction. When maternal contamination is suspected, we propose that at least 30 metaphases from different slides should be counted. This could yield fetal as well as maternal chromosomes.

Gorlin-Chaudhry-Moss syndrome revisited: expanding the phenotype.

Gorlin–Chaudhry–Moss syndrome (OMIM 233500) is a rare congenital malformation syndrome with the cardinal manifestations of craniofacial dysostosis, hypertrichosis, underdeveloped genitalia, ocular, and dental anomalies. Since 1960, only six affected individuals have been reported. We report a 4‐year and 6‐month‐old female patient with this phenotype and review the clinical presentation of all patients known so far. Previously unreported malformations of the extremities, larynx, and nose are also described, expanding the phenotype of this rare syndrome. Array‐CGH analysis did not show pathological deletions or duplications.

PRENATAL DIAGNOSIS OF DIGEORGE SYNDROME

Velocardiofacial syndrome, conotruncal congenital heart disease and DiGeorge syndrome present different aspects of the same clinical situation, but congenital heart disease is common to them all [1]. Other defects found in these syndromes include abnormal facies, thymic hypoplasia, cleft palate, and hypocalcemia. A deletion at chromosome 22q11 is the second most common chromosomal defect detected in patients with congenital heart disease. Although familial inheritance occurs, most cases are sporadic [2]. A patient aged 29 years, gravida 2, para 1, presented to our outpatient clinic with an initial diagnosis of polyhydramnios at the 33 rd week of gestation. Her medical history revealed that her first delivery was at term by cesarean section, but the baby died in infancy of cyanotic heart disease, diffuse bronchopulmonary disease, and infection. Postmortem genetic investigation revealed that the neonate had a 22q11 deletion, with associated cardiovascular anomalies. The parents were genetically normal. Obstetric examination determined that the patient was at 33 weeks of gestation, and fetal biometric measurements suggested a gestational age of 31–32 weeks. The amniotic index was 18 cm and mild polyhydramnios was detected. Systematic ultrasonographic investigation revealed a ventricular septal defect, a secundum-type atrial septal defect, pulmonary atresia, and an anomaly of the overriding aorta. These findings were interpreted as components of tetralogy of Fallot (TOF). The patient had not attended the hospital for prenatal screening during her second trimester. Cordocentesis was performed for prenatal diagnosis. The cytogenetic analysis was evaluated as normal, but considering the clinical findings and the patient’s prior history, locus-specific fluorescence in situ hybridization (FISH) was planned to detect possible DiGeorge syndrome. A Tuple1-Hira

Methylation of SOCS3 in Myeloproliferative Neoplasms and Secondary Erythrocytosis/Thrombocythemia

Objective: Myeloproliferative neoplasms (MPNs) like essential thrombocythemia (ET), polycythemia vera (PV), and primary myelofibrosis (PMF) are acquired clonal hematopoietic stem cell disorders and originate from a multipotent hematopoietic stem cell. The SOCS1 and SOCS3 genes are negative regulators of the JAK/STAT signal pathway. In this study we investigate the promoter methylation of these genes in the pathogenesis of MPNs and secondary erythrocytosis/thrombocythemia. Materials and Methods: Promoter methylation of SOCS1 and SOCS3 genes was analyzed with methylation-specific PCR. PCR products were analyzed by agarose gel electrophoresis. Results: No disease-specific CpG island methylation of SOCS1 was observed. Hypermethylation of the SOCS3 promoter was identified in 5 out of 19 (26.3%) PV cases, 2 out of 21 (9.5%) ET cases, 1 out of 5 (20%) PMF cases, and 9 out of 42 (21.4%) cases of secondary erythrocytosis/thrombocythemia. Conclusion: The results revealed that promoter methylation of the SOCS3 gene suggests a possible role for SOCS3 methylation in the pathogenesis of MPNs and secondary erythrocytosis/thrombocythemia. Conflict of interest:None declared.

The presence of MEFV gene mutations in patients with primary osteoarthritis who require surgery

Background/Aims Chronic arthritis of familial Mediterranean fever (FMF) involves weight-bearing joints and can occur in patients without a history of acute attack. Our aim was to investigate a possible causal relationship between FMF and osteoarthritis in a population in which FMF is quite common. Methods Patients with late stage primary osteoarthritis were enrolled, and five MEFV gene mutations were investigated. The frequency of MEFV gene mutations was compared among patients with osteoarthritis and a previous healthy group from our center. Results One hundred patients with primary osteoarthritis and 100 healthy controls were studied. The frequency of MEFV gene mutations was significantly lower in the osteoarthritis group (9% vs. 19%). M694V was the most frequent mutation (5%) in the osteoarthritis group, whereas in the control group, E148Q was the most common (16%). In subgroup analyses, the mutation frequency of patients with hip osteoarthritis was not different from that of patients with knee osteoarthritis and controls (7.1%, 9.7%, and 19%, respectively). There were no differences among the three groups with respect to MEFV gene mutations other than E148Q (8.1% vs. 3.6%). E148Q was significantly lower in the osteoarthritis group than in the controls (16% vs. 1%), although the mutations did not differ between patients with knee osteoarthritis and controls. Conclusions In a population with a high prevalence of MEFV gene mutations, we did not find an increased mutation rate in patients with primary osteoarthritis. Furthermore, we found that some mutations were significantly less frequent in patients with osteoarthritis. Although the number of patients studied was insufficient to claim that E148Q gene mutation protects against osteoarthritis, the potential of this gene merits further investigation.

A new syndrome associated with absence of lower lid lacrimal punctum, ptosis, elevation deficiency of both eyes and mild facial dysmorphism.

A considerable volume of literature has been published on the association of lacrimal outflow dysgenesis with developmental anomalies or systemic syndromes. We report three affected individuals in a consanguineous family those are associated with bilateral ptosis, upper ocular movement limitation, and absence of the lacrimal punctum. T our knowledge, this is the first article reporting the association of bilateral ptosis, facial dysmorphism, upper ocular movement limitation, and absence of the lacrimal punctum in a hereditary form. As a sole example, these findings may be accepted as a new syndrome with autosomal recessive pattern because of consanguinity.

Rare chromosomal complement of trisomy 21 in a boy conceived only by IVF.

In the article ‘Rare chromosomal complement of trisomy 21 in a boy conceived by IVF and cryopreservation’ presented in Reproductive BioMedicine Online (Quiroga et al., 2009) a case of mosaic Down syndrome with an unusual karyotype was reported. The chromosomal complement consisted of two different cell lines, one predominantly trisomic with a derivative chromosome due to a Robertsonian translocation (21;21) and another carrying a ring chromosome 21. The assisted reproductive techniques and cryopreservation were held responsible in this rare type of mosaic Down syndrome case. This letter presents a similar case with three different cell lines including predominantly a ring 21 chromosome, a Robertsonian translocation (21;21) and a monosomy 21 (Figure 1). The karyotype was 46,XY r(21) [87], 45,XY, 21 [21], 46,XY, t(21;21)(q10;q10) [6] (Figure 1). This one-month old case was born to a non-consanguineous family as the first child. His mother was 29 years old and his father was