Zerrin Yilmaz

Chromosome 22q11.2 deletion and phenotypic features in 30 patients with conotruncal heart defects.

This report describes the dysmorphic features and frequency of 22q11.2 deletion (del22q11) in 30 Turkish patients with conotruncal heart defects (CTHDs). Fluorescence in situ hybridization (FISH) analysis revealed deletions in the 22q11.2 region in nine (30%) individuals. The CTHDs in this group were tetralogy of Fallot (four cases), double‐outlet right ventricle (DORV) (two cases), transposition of great arteries (two cases), and ventricular septal defect (VSD) associated with other CTHDs (one case). The frequency of del22q11 in the study group was relatively high because many of the patients with dysmorphic findings also had cardiac anomalies involving the pulmonary artery, ductus arteriosus, or the aortic arch and its main branches. Twenty of the 30 patients exhibited several dysmorphic findings. Two of the nine patients with del22q11 exhibited no apparent dysmorphic features other than sacral dimple. Interestingly, one of the patients with del22q11 had a phenotypic appearance similar to that seen in oculo‐auriculo‐vertebral spectrum (OAVS). This individual had left microtia, atresia of the external meatus, mandibular asymmetry, and peripheral facial nerve paralysis. His mental development was normal and there were no abnormalities on ophthalmological examination. The CTHDs in this patient were situs inversus dextrocardia, DORV, pulmonary stenosis, and VSD. Radiographs of this patient showed platybasia, complete fusion of C2–C3, and posterior fusion of the T1‐T2 vertebrae. This particular case indicates that the phenotypic features of del22q11 and OAVS may overlap.

Chromosome heteromorphisms: an impact on infertility

IntroductionCytogenetic heteromorphisms are described as heritable variations at specific chromosomal regions without a proven impact on phenotype.Materials and methodsWe compared the presence of chromosome heteromorphisms in the karyotypes of two patient groups. The first group of patients consisted of 276 individuals of 138 infertile couples. The second group, consisted of 1,130 amniocentesis samples. This group was considered to be a sample of the fertile population, as the fetus being karyotyped is the result of a spontaneous pregnancy. Fetal karyotyping was made due to the standard indications for prenatal diagnosis, such as abnormal maternal serum screening results. Results and discussionEighteen infertile patients (6.52%) and twenty fetuses (1.77%) were found to have chromosome heteromorphisms. The difference between the two groups was statistically significant (p < 0.0001).ConclusionThese results are consistent with other similar studies that suggest the yet undefined relationship between chromosome heteromorphisms and infertility.

Conventional and molecular cytogenetic findings of myelodysplastic syndrome patients.

AbstractMyelodysplastic syndrome (MDS) involves myeloid cells of the bone marrow, which is important in progressive bone marrow insufficiency. Of all MDS patients, 40%–50% have at least one chromosomal rearrangement. Loss of specific chromosomal regions like 5q– and 7q– are usually the secondary cytogenetic abnormalities associated with MDS. In order to detect chromosome abnormalities associated with MDS, bone marrow samples from 26 patients diagnosed as MDS were obtained prior to chemotherapy. Both conventional cytogenetic analyses and fluorescence in situ hybridisation (FISH) methods were performed and locus–specific probes for 5q and 7q were used. Results obtained were compared. Twenty–one patients had normal karyotypes and four patients had abnormal karyotypes, while in one patient we could not obtain metaphases from cultures. Three patients with normal karyotypes revealed del (5q), two patients had del (7q) and one patient had monosomy (7). A total of 10 of 26 patients had chromosome changes visualised by either conventional or molecular cytogenetics (~38.5%). Our results show that both methods are important in diagnosis and follow up of MDS patients. When used together, conventional cytogenetics and FISH detect clinically significant chromosome abnormalities in MDS patients.

Familial cystic nephroma in two siblings with pleuropulmonary blastoma

Cystic nephroma (CN) and pleuropulmonary blastoma (PPB) are rare tumors. In the cases presented here, a 13-month-old boy underwent right radical nephrectomy for CN. From the family history we learned that four years ago the patient’s older sister underwent left radical nephrectomy for CN at a different center when she was 4 years old. A lung tumor was detected in the sister one year after nephrectomy. Biopsy from the lung tumor revealed PPB, and the sister died within one year after biopsy. To the knowledge of the authors, these cases represent the second reported familial occurrence of CN and the fourth of CN and PPB.

Fluorescent in situ hybridization studies in multiple myeloma.

Abstract Conventional cytogenetic analysis and fluorescence in situ hybridization (FISH) results of bone marrow samples of 36 multiple myeloma (MM) patients at the time of diagnosis have been evaluated. Three probes for chromosome 13q (RB1, D13S319, D13S25), one for 14q32 (IgH) and one for 17p13 (p53) have been used for hybridization with fixed cells. Twenty patients (55·5%) had normal karyotypes, whereas eight (22·2%) had numerical or structural chromosomal abnormalities. We did not find metaphases for chromosome analysis in eight (22·2%) patients. Fluorescence in situ hybridization analyses revealed at least one or more abnormal results in 25 (69·5%) cases, whereas 11(30·5%) cases had no abnormal findings. 14q32 rearrangement was the most common finding in FISH analyses and has been detected in 21 cases (58·3%). 13q deletion and 17p deletion have been detected in 11 (30·5%) and 5 (13·9%) cases, respectively. Fluorescence in situ hybridization studies including 14q32 and 17p13 chromosome regions may yield quite significant results during clinical follow-up of MM.

Influence of recasting different types of dental alloys on gingival fibroblast cytotoxicity

STATEMENT OF PROBLEM Surplus alloy from the initial casting is commonly reused with the addition of new alloy. This recasting procedure could affect the cytotoxicity of dental alloys. PURPOSE The purpose of this in vitro study was to evaluate the effect of repeated casting of high-noble and base metal alloys on gingival fibroblast cytotoxicity. MATERIAL AND METHODS Disk-shaped specimens (5 × 2 mm, n=60) of a high-noble (Au-Pt) and 2 base metal (Ni-Cr and Cr-Co, n=20) alloys were prepared with 100% new alloy and 50%, 65%, and 100% once recast alloy. The elemental composition of specimens was analyzed with X-ray energy-dispersive spectroscopy. Five specimens from each group were conditioned in saline with 3% fetal bovine serum albumin. The conditioning media were analyzed for elemental release with atomic absorption spectroscopy. Cytotoxic effects were assessed on human gingival fibroblast with a 3-(4.5-dimethylthiazol-2-yl)-2.5-diphenyl tetrazolium bromide (MTT) colorimetric assay. The data were analyzed with 1-way and 2-way ANOVA and Tukeys HSD multiple comparison test (α-=.05). RESULTS Elemental compositions of Co-Cr and Au-Pt alloys were significantly different among casting protocols. Elemental release of Co-Cr and Ni-Cr alloys was significantly different between new and recast specimens (P<.001). Nickel release increased with recast alloy addition. The 2-way ANOVA showed a significant effect of the casting procedure (P<.001) alloy group (P<.001) and their interaction for cytotoxicity (P<.001). The Ni-Cr alloy groups with 65% and 100% recast alloy had lower cellular activity than all other specimens (P<.001). CONCLUSIONS The results of this study indicated that alloys containing nickel have increased cytotoxic effects and that composition of the alloys affected the cytotoxicity. Furthermore, recasting nickel-containing alloys with 65% surplus metal addition significantly increased the cytotoxic activity.

Determination of HER-2/Neu Status in Hepatocellular Carcinoma Cases

Specific chromosome abnormalities and genetic changes in hepatocellular carcinoma (HCC) have been demonstrated by conventional cytogenetic studies or molecular cytogenetic approaches like comparative genomic hybridization and loss of heterozygosity analyses. HER-2/Neu amplification and expression has been studied as a molecular target for treatment of HCC, and there are conflicting results. We aimed to determine HER-2/Neu status in archive materials of HCC patients by fluorescence in situ hybridization (FISH). Among the 35 patients, 2 had HER-2/Neu amplification and 3 had increased chromosome 17 copy number. All these patients had grade 2 or 3 tumor with a diameter of 3-12 cm. We conclude that although HER-2/Neu amplification is not the primary mechanism in the development of liver tumors, it might play a role in one of the steps of multistage carcinogenesis.

Long-term colchicine therapy in a patient with Behçet's disease and acute promyelocytic leukemia

Behçet’s disease causes a continuous T-lymphocytic mediated inflammatory reaction in the small arterioles, which results in gradual destruction of any human organ or system. The benefit of treatment with colchicine in patients with Behçet’s disease has been reported in literature. Acute leukemia has seldom been associated with Behçet’s disease, although acute promyelocytic leukemia is a particular subtype of leukemia that is often characterized by special cytogenetic abnormalities. We report a male patient with acute promyelocytic leukemia and Behçet’s disease who had received long-term treatment with colchicine. To our knowledge, this is the first report of the concomitant occurrence of acute promyelocytic leukemia and Behçet’s disease, which suggests that long-term colchicine therapy has a role in the pathogenesis of acute promyelocytic leukemia. The patient described has been treated with retinoic acid and idarubicin (the ATRA-IDA protocol). At the time of this writing, his disease is in clinical remission.

Chromosome 1p36 and 22qter deletions in paraffin block sections of intracranial meningiomas.

Meningiomas are the most frequent benign tumors of the intracranial cavity. The classification and underlying pathogenetic mechanisms have been reported to be investigated by both pathological and genetic methods. In this study, we aimed to detect Ip36 and 22qter deletions by fluorescence in situ hybridization (FISH) in archival materials of 50 intracranial meningioma patients. The clinical material consisted of paraffin-embedded tissue sections from 50 patients who were surgically treated and had histopathologic diagnosis of an intracranial meningioma. We observed Ip36 deletion in 23/50 (46%) and 22qter deletion in 33/50 (66%) patients. In addition, we observed 22qter deletion in 26/36 (72.2%) patients with meningothelial meningioma. This finding implies that 22qter deletion might play an important role in the pathogenesis of meningothelial meningioma. On the other hand, no alterations were documented in the frequency of these chromosomal alterations according to the grade of meningiomas, suggesting that malignant progression of these tumors depends on other, more relevant, genetic changes.

Comparison of the Results of PCR-RFLP and Reverse Hybridization Methods Used in Molecular Diagnosis of FMF

Familial Mediterranean fever (FMF) is characterized by recurrent fever, serositis, and arthritis. Due to the abundance of mutations and clinical heterogeneity of the disease, different screening methods have been developed. In this study, we aimed to compare our findings of mutations determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) with reverse hybridization (RH) methods. In 152 of 263 patients (57.79%) different mutations were determined with RH. Allelic frequencies were E148Q 6.84%, M680I(G/C) 3.61%, M694V 20.91%, V726A 7.03%, P369S 1.33%, F479L 0.19%, M680I(G/A) 0.76%, M694I 0.57%, K695R 0.57%, A744S 0.38%, R731H 0.38%, and del1692 0%. Frequent mutations were also confirmed by PCR-RFLP. There were no conflicting results between the two methods. Four of these genotypes were homozygous for a single mutation, 15 were heterozygous for two mutations, 8 were heterozygous for a single mutation, 1 was heterozygous for three mutations, and 1 was homozygous for one mutation and heterozygous for another mutation. It has been reported that analytical sensitivity of RH is 97%. We did not find a discrepancy between the two methods. In 21 patients, we detected additional mutations with RH. This finding was regarded as an advantage of RH, and we concluded that this assay is a useful method for detection of first stage FMF mutation screening.