Partial deletion of the long arm of chromosome 13 (q32q33.2) associated with mental retardation, choanal atresia and fish mouth.

13q deletion syndrome is characterized by mental and motor retardation, craniofacial dysmorphic facial appearance and various congenital malformations. In this article, we present a new case with 13q deletion syndrome phenotypically characterized by fish mouth, choanal atresia and severe mental and motor retardation. In order to determine the certain localization of deleted region high resolution multicolor-banding technique was performed and the karyotype determined as 46,XX,del(13)(q32q33.2). To come in future to a genotype-phenotype correlation, it is very important to delineate the deleted region in such cases in detail by cytogenetic/ molecular cytogenetic methods.

A prenatally sonographically diagnosed conotruncal anomaly with mosaic type trisomy 21 and 22q11.2 microdeletion/DiGeorge syndrome.

A prenatally sonographically diagnosed conotruncal anomaly with mosaic type trisomy 21 and 22q11.2 microdeletion/DiGeorge syndrome: We report a prenatally sonographically diagnosed conotruncal and urogenital anomaly. Postnatally, the patient presented with seizures, hypocalcemia, hypoparathyroidism and thymic aplasia and diagnosed as DiGeorge syndrome. Echocardiography showed malalignment VSD, supravalvular pulmonary stenosis and overriding aorta. Chromosome and FISH studies showed the association of mosaic type trisomy 21 and 22q11.2 microdeletion. The present patient is the second case of mosaic type of Down syndrome associated with 22q11.2 microdeletion. In addition the patient also had clinical and laboratory features of DiGeorge syndrome.

Partial trisomy (11;22) syndrome with manifestations of Goldenhar sequence due to maternal balanced t(11;22).

Here we report a 15-year-old girl patient who had severe mental and growth retardation, cleft palate, hemifacial microsomia, skin tags, hypoplasia of the external auditory canal, scoliosis and renal agenesis. Our patient was the fourth child of nonconsanguineous marriage. Peripheral blood chromosomal analysis of the patient revealed 47,XX,+der(22)t(11;22)(q23;q11). The maternal karyotype was reported as 46,XX,t(11;22)(q23;q11). Maternal balanced translocation t(11;22)(q23;q11) causing Goldenhar syndrome with 47,XX,+der(22) has not been reported previously. The presented case clearly indicates that in every case with Goldenhar syndrome, chromosome analysis should be done for the possibility of unbalanced translocations.

Relationship between embryo quality and aneuploidies.

Many high-grade embryos selected for transfer according to their morphological evaluation were detected to have chromosomal abnormalities after aneuploidy screening for infertility by preimplantation genetic diagnosis (PGD). The aim of this study was to detect if there is any correlation between embryo quality and genetic status. The chromosomal status of the day three embryos was studied by multicolour fluorescence in-situ hybridization for chromosomes 13, 18, 21, X and Y. PGD was performed on 132 patients for 1107 embryos. The correlation between embryo quality and aneuploidy was analysed. The analysis showed that a large proportion of normal embryos (50.7%, n = 280) were grade I. In addition, a considerably high proportion of aneuploid embryos (36.1%, n = 83) were evaluated as grade I. There was a significant relationship between PGD results and embryo grades (P = 0.001). Of the 69 polyploid embryos, 21.7% were grade I and 37.8% were grade II. Of the 83 haploid embryos, 27.8% were grade I and 34.9% were grade II. Euploidy was positively related to morphological grade of embryo (P = 0.001). It was also possible for chromosomally abnormal embryos to have a good developmental potential, and they could be selected for embryo transfer unless the PGD procedure was applied.

Popliteal pterygium associated with neonatal Marfan syndrome: case report.

This report describes the case of a male infant with neonatal Marfan syndrome who also exhibited popliteal pterygia. The patient's father had classic Marfan syndrome. The differential diagnosis in the neonatal case included congenital contractural arachnodactyly (Beals syndrome) and various forms of popliteal pterygium syndrome. We note the diagnostic features of the case, discuss the novel finding of pterygia in association with neonatal Marfan syndrome, and highlight the possible role of collagen defects in the pathogenesis of limb pterygia.

Evaluation of DNA damage using the comet assay in children on long-term benzathine penicillin for secondary prophylaxis of rheumatic fever.

BACKGROUND: Benzathine penicillin is the most widely used antibiotic in the prophylaxis of children with rheumatic fever. The aim of the present study was to evaluate the DNA damage in children receiving one dose of 1.2 million units benzathine penicillin every 4 weeks over a long period to prevent recurrences of rheumatic fever. METHODS: Thirty-five children with confirmed rheumatic fever under benzathine penicillin prophylaxis were enrolled in the study and 35 healthy children with similar ages and socioeconomic backgrounds served as controls. To detect any DNA damage, the comet assay was performed on circulating lymphocytes from the study subjects. RESULTS: Damaged (limited and extensive migration) cells in children on prophylactic therapy were higher than those in controls (P<0.001). CONCLUSIONS: It has been suggested that differences in the comet scores were induced by the administration of benzathine penicillin over a long period of time and further investigations are needed to confirm this toxic effect.

Sister-chromatid exchange analysis on long-term benzathine penicillin for secondary prophylaxis of rheumatic fever.

A single intra-muscular injection of 1.2 millions units of benzathine penicillin every 4 weeks is the most widely used method for the antibiotic prophylaxis of rheumatic fever. The aim of this study is to evaluate the effect of long-term benzathine penicillin on DNA in patients with rheumatic fever. Thirty children with confirmed rheumatic fever who were on the benzathine penicillin prophylaxis were enrolled in the study, and 30 similar normal children served as a control group. To detect any DNA damage, SCE analysis were performed in circulating lymphocytes of the subjects. A statistically significant increased frequency of SCE was observed in children on the benzathine penicillin prophylaxis (no = 30, mean SCEs/cell +/- SD 7.54 +/- 1.81) as compared to a control group (no = 30, mean SCEs/cell +/- SD 5.82 +/- 1.40). It has been suggested that the difference in the SCE frequencies was induced by the administration of the benzathine penicillin for a long time, and further investigations are needed to confirm this toxic effect.

Non-invasive early prenatal diagnosis using fluorescent in situ hybridization on transcervical cells: comparison of two different methods for retrieval.

OBJECTIVE: We compared the efficiencies of uterine and endocervical lavage to retrieve fetal cells from first trimester pregnancies for further analysis with fluorescent in situ hybridization (FISH). STUDY DESIGN: Transcervical cell (TCC) samples were collected at 7-10 weeks of gestations by uterine lavage (13 women) and by endocervical lavage (12 women) who were scheduled for volunteer termination of pregnancy. A sample of placenta was also obtained for cytogenetic analysis to confirm the sex or genotype in the end of the procedure. FISH was performed using probes for the chromosomes 18, X and Y in a three color hybridization protocol. The statistical analysis included chi(2)-analysis, and t-test. RESULTS: Sufficient cells were obtained in 12 of the 13 (92.3%) in uterine lavage and 10 of the 12 (83.3%) in endocervical lavage group for FISH procedures for fetal sex prediction. The mean success rate of signal detection for FISH procedure was 91.7% (range 83-97%). Fetal sex was correctly predicted in 11 of 12 (91.6%) with uterine lavage and 8 of 10 (80.0%) in endocervical lavage and the difference was statistically insignificant. CONCLUSION: This study demonstrated that there are available cells of fetal origin in the lower part of the uterus and these cells may be collected successfully as early as 7 weeks of the gestation. In addition, our results show that endocervical lavage method is as effective as uterine lavage. FISH has been successfully used to detect status of aneuploidy and sex of the fetus from TCC.

Retinal vein occlusion and factor V Leiden and prothrombin 20210 G:A mutations.

PURPOSE: The aim of this study was to establish whether the factor V Leiden mutation and the prothrombin 20210 G:A mutation were risk factors for retinal vein occlusion. METHODS: Blood samples were obtained from 40 patients with retinal vein occlusion and from 50 healthy normal volunteers as controls. Polymerase chain reaction assays were done to detect factor V Leiden and prothrombin 20210 G:A mutations, and the two groups were compared. RESULTS: Two (5%) of 40 patients with retinal vein occlusion and three (6%) of 50 controls were heterozygous for factor V Leiden (p=0.84). None of the individuals in either group had the prothrombin 20210 G:A mutation. CONCLUSIONS: There was no significant association between retinal vein occlusion and the factor V Leiden mutation.

A comparative study of genotoxic effects in the treatment of trichomonas vaginalis infection: metronidazole or nalidixic acid.

We performed a prospective randomized study to compare the potential genotoxic effects of metronidazole and nalidixic acid which they are used in the treatment of Trichomonas vaginalis infection. 20 patients with Trichomonas vaginalis infections participated in this study. 14 patients with vaginal trichomoniasis were treated with therapeutic doses of metronidazole 250 mg 3 times/d and six patients were treated with nalidixic acid 400 mg twice a day for 10 d. The genotoxic potential of a variety of mutagenic and carcinogenic agents can be evaluated by sister-chromatid exchange (SCE) test as a rapid cytogenetic test. An increased number of exchanges in lymphocytes reflects the influence of mutagens. No significant difference was observed in the SCE frequency of metronidazole treated patient however, a statistically significant increase (P<0.05) after nalidixic acid treatment could be described. We conclude that in spite of wide use of nalidixic acid for Trichomonas vaginalis infection, because of its potential genotoxic effect its usage must be individualized especially for pregnant women and small babies.

Sister-chromatid exchange frequencies in postmenopausal hormone replacement patients.

Our objective was to evaluate the frequency of sister-chromatid exchange (SCE) during hormone replacement therapy in postmenopausal women. Thirty-four asymptomatic postmenopausal women with a minimum 12 months since last menstrual period and surgical menopausal women were included in the study. Seventeen patients who were in spontaneous menopause were administered conjugated estrogen and medroxyprogesterone acetate (group A), and the others who were in surgical menopause were given 17beta-estradiol only (group B). Peripheral lymphocytes were obtained at the beginning and at the end of the third month of therapy. The mean age of the patients was 50. 67+/-4.79. There were statistically significant differences in terms of SCE frequencies between pre- and posttreatment levels of both groups (p<0.001 and p=0.003, respectively). It is likely that estrogens with or without progesterone have an effect in increased SCE frequency and this issue may be an evidence for the increased potential for malignancies.

Detection of chromosomal abnormalities by fluorescent in-situ hybridization in immotile viable spermatozoa determined by hypo-osmotic sperm swelling test.

If randomly selected immotile spermatozoa are used for intracytoplasmic sperm injection (ICSI), pregnancy rates are significantly decreased. The hypo-osmotic swelling test (HOST) is the only method available to detect the viable, but immotile spermatozoa for ICSI. However, evidence is still lacking for the chromosomal abnormalities for the normal-looking, but immotile spermatozoa positive for HOST. Sperm samples from 20 infertile men with normal chromosomal constitution were obtained. After Percoll separation, morphologically normal but immotile spermatozoa were transported individually into HOST solution for 1 min using micropipettes. Cells that showed tail curling with swelling in HOST were then transferred back into human tubal fluid solution to allow reversal of swelling. These sperm cells were fixed and processed for the multi-colour fluorescence in-situ hybridization (FISH) for chromosomes X, Y and 18. The same FISH procedure was applied for the motile spermatozoa from the same cohort, which formed the control group. The average aneuploidy rates were 1.70 and 1.54% in 1000 HOST positive immotile and motile spermatozoa respectively detected by FISH for each patient. Our results indicate that morphologically normal, immotile but viable spermatozoa have an aneuploidy rate similar to that of normal motile spermatozoa.

Sister chromatid exchange rate from pleural fluid cells in patients with malignant mesothelioma.

This study assessed the impact of malignant mesothelioma on the frequencies of sister chromatid exchange (SCE) in the pleural effusion cells. Ten patients with mesothelioma and 20 control subjects were included in the study. The control subjects were the patients with tuberculosis pleurisy, and the remaining 10 subjects of control group were healthy volunteers and only heparinized blood samples were collected from these subjects. The pleural effusion cells were cultured with conventional culture methods. The samples were obtained from the patients after histopathologic confirmation of the malignancy but before the initiation of chemotherapy or radiotherapy. At the end of the culture period and 48 h prior the harvesting, BrdU was added into flasks. Totally, 100 metaphases were scored for each sample. In this study, we found that the SCE frequencies of malignant pleural mesotheliomas were significantly higher than the control subjects (P<0.001). Six of 10 patients came from central Anatolia, which is of great importance due to high rate of exposure to asbestosis in this region.

A case associated with Walker Warburg syndrome phenotype and homozygous pericentric inversion 9: coincidental finding or aetiological factor?

We describe a rare occurrence of pericentric inversion in homologues of chromosome 9 observed in a 2-mo-old female baby with eye and brain abnormalities. Her clinical and neuroradiological features are similar to the signs of Walker-Warburg syndrome. We found the same inversion in heterozygous condition in all metaphases of both parents, who are related, and in two grandparents and their mother. The cytogenetic abnormality alone does not explain the phenotype in this patient, but it warrants further linkage studies with emphasis on the pericentric region of chromosome 9 in patients with Walker-Warburg syndrome phenotype. This family case is unique and raises suspicions about whether the pericentric region of chromosome 9 has any connection with the phenotype of Walker-Warker syndrome.

Clinical assessment of EMA/CO induced DNA damage in peripheral blood lymphocytes of high-risk gestational trophoblastic tumor patients.

From 1989 to 1994, Etoposide, Methotrexate, Actinomycin-D, Cyclophosphamide, Vincristine, Folic acid (EMA/CO) regimen was administered to seven patients with high-risk gestational tumours according to the Bagshawe 1976 criteria. Peripheral blood lymphocytes were obtained from two of these seven high-risk gestational trophoblastic patients receiving the EMA/CO regimen, and damage levels of DNA during chemotherapy were assessed using SCGE (single cell gel electrophoresis) assay. Additionally, the efficacy, toxicity and clinical results of EMA/CO regimen were evaluated in patients with high-risk gestational trophoblastic tumours. Fever (71.4%), leukopenia (57%), increase in transaminase concentrations (57%), trombocytopenia (57%), and anemia (57%) were among the most frequent side-effects of the EMA/CO regimen. All these toxic effects were reversible and there was no need to stop the therapy. EMA/CO is highly effective in patients with high-risk gestational trophoblastic disease and its toxicity is predictable and reversible. Because of chemotherapy, DNA damage that is shown in peripheral blood lymphocytes, increases at the 8th day of the EMA/CO regimen. When DNA damage is higher in patients, the course of chemotherapy per each patient is shortened. When DNA damage is higher in the patients, the multisystem effects due to toxicity are more significant. The SCGE assay has many possibilities in such research and has proved to be a relatively simple, quick and sensitive technique.