RARß gene methylation is a candidate for primary glioblastoma treatment planning.

BACKGROUND: We screened RARß methylation in primary glioblastoma multiforme (GBM) and the results were evaluated based on the clinical data and treatment type. OBJECTIVE: The objective of this study was to find new areas for the usage of MS-HRM applications in the determination of methylation levels in primary GBM samples and it shows the association of RARß methylation with the clinical outcome. METHODS: In our study, tumor samples were collected during surgical resection by the Department of Neurosurgery. The clinical and radiologic data was carefully reviewed, compared, and evaluated with the histological results. The methylation status of RARß was determined by using MS-HRM. RESULTS: RARß gene methylation was detected in 24 out of 40 cases (60%), with different quantitative methylation levels. The mean survival time was 19 months form ethylated cases and 15 months for the non-methylated cases. The survival time of the patients who received treatment was 25 months and the survival time of the patients who received radiotherapy alone or where no treatment protocol applied was 15-20 months. Therefore, a significant difference in survival rates has been observed (P<0.05). This study indicates a potential prognostic value for GBM treatment planning. CONCLUSION: Our study is the first study to investigate RARß methylation in primary GBMs. We conclude that the RARß gene could be a new prognostic and predictive candidate marker to designate the treatment protocol for primary GBMs.

Herpes zoster ophthalmicus reactivation following maxillary sinus lift operation: A case report.

PURPOSE: To present a case of Herpes Zoster Ophtalmicus (HZO), which was reactivated postoperatively after a sinus lift operation. MATERIALS AND METHODS: A 39-year-old male was referred to our clinic for implant-supported dental rehabilitation. He had maxillary missing teeth in positions 13, 14, 15 and 16 and a pneumatised right maxillary sinus with a bone height of 2 mm. Lateral sinus lifting and bone block grafting was performed before implant insertion. Twelve days after the sinus lift, the patient complained of pain and itching at the infraorbital area extending to the forehead. Clinical examination revealed no signs of infection or allergy. The patient received consultation from a dermatologist in order to rule out a possible dermatological disorder. Finally he was diagnosed with HZO. RESULTS: HZO was managed with systemic acyclovir treatment. Vesicular rashes and ptosis was seen 3 days after the medical treatment. After 1 month no postoperative skin or orbital sequela was seen. Three implants were inserted at the right posterior maxilla 5 months after sinus lift. One-year followup was uneventful. CONCLUSIONS: Dermatological diseases should always be kept in mind during the differential diagnosis of orofacial pain. In this case the proximity of the operation site and affected area gave rise to the idea that surgical trauma had a possible role in the reactivation of the virus. However, the process of reactivation is not entirely understood and requires further investigations. Early diagnosis is essential for HZO in order to avoid debilitating complications such as postherpetic neuralgia and blindness.

The effect of hemostatic agents and tissue adhesive on injured peripheral nerve healing in rats - Part I. Electrophysiological study.

BACKGROUND: In the practice of maxillofacial surgery, bleeding and nerve injury have common problems. In the control of bleeding, hemostatic agents and tissue adhesives have been frequently used. The effect of these hemostatic agents and tissue adhesives on the injured neural tissues has not been known. OBJECTIVES: In this study, we aimed to investigate the effects of hemostatic agents and tissue adhesive on injured nerve tissues. MATERIAL AND METHODS: Forty-two rats randomly divided into seven groups: Control, Oxidized Regenerated Cellulose (ORC), Gelatine Sponge (GS), Bovine Collagen (BC), Ankaferd BloodStopper (ABS), Glutaraldehyde Surgical Adhesive (BioGlue®) and N-butil-2 cyanoacrylate (Glubran®2). The left sciatic nerves were crushed and surrounded by hemostatic agents and tissue adhesives. At the end of 12 weeks, the surgical site was reopened and electrophysiological recordings were performed. RESULTS: In the ORC, GS, and BC groups, the compound action potential (CAP) values were lower compared to the control group (p < 0.05). Although the values of CAP in the ABS group were higher than in the control group while CAP values in the BioGlue and Glubran®2 groups were lower than the control group, there was no statistical significance between the experimental and control groups (p > 0.05). In the ORC, BC, GS, and Glubran®2 groups, the nerve conduction velocities (NCV) values were lower than in the control group (p < 0.05). In the ABS and BioGlue groups, NCV values were lower compared to the control group but no significant differences were found (p > 0.05). CONCLUSIONS: The present study provides evidence that ABS is the most suitable hemostatic agent due to its favorable effect on the healing of injured neural tissues. BioGlue is also a suitable surgical agent with no adverse effects.

IDH1 mutations is prognostic marker for primary glioblastoma multiforme but MGMT hypermethylation is not prognostic for primary glioblastoma multiforme.

PURPOSE: To establish the frequency of IDH1 mutations and MGMT methylation in primary glioblastomas. EXPERIMENTAL DESIGN: We screened primary glioblastoma multiforme (GBM) in a population-based study for IDH1 mutations and MGMT methylation and correlated them with clinical data. RESULTS: IDH1 mutations were detected in 5 of 40 primary glioblastomas (12,5%). Primary GBM patients carrying IDH1 mutations were significantly younger, mean age of 41±5.06years, than patients with wild-type IDH1, mean age of 57±2,29years, p=0.011. The mean survival time of all GBM patients with and without IDH1 mutations was 19months (5 cases) and 16months (35 cases), respectively (p>0,05). MGMT methylation was detected in 13 of the 40 patients (32,5%). MGMT-promoter methylation did not correlate with overall survival (OS; p>0,05). CONCLUSION: In summary, our study is the first study to investigate the IDH1 mutation status and MGMT methylation in primary GBMs in Turkish population and confirmed IDH1 mutation as a genetic marker for also primary GBMs. Our data are still insufficient for definite ascertainment; and our preliminary results suggest: IDH1 status shows an association with younger age and there is a lack of association between IDH1 mutation and survival time. Furthermore MGMT promoter methylation had no prognostic value and lower frequency in primary glioblastomas.

Propolis accelerates the consolidation phase in distraction osteogenesis.

We evaluated the effect of propolis on new bone formation after distraction osteogenesis (DO). This study examined 3 groups: control group, P100, and P200. Rabbits underwent DO of the left mandible after an osteotomy between the first molar and the mental foramen. Bone mineral content and bone mineral density were evaluated using dual-energy x-ray absorption 1 and 4 weeks after the procedure. The volume of connective tissue and new bone and the number of capillaries were measured using stereologic analysis after the subjects were killed. Dual-energy x-ray absorption showed that the bone mineral content and bone mineral density were higher in the groups treated with propolis by week 4, and these parameters were higher in the P200 group. Stereologic analysis showed no significant differences in connective tissue volume and number of capillaries among the groups. New bone volume was lowest in the P200 group. We concluded that propolis accelerates bone formation and may shorten the consolidation phase with DO.

Investigation of key miRNAs and target genes in bladder cancer using miRNA profiling and bioinformatic tools.

Despite the association of several miRNAs with bladder cancer, little is known about the miRNAs' regulatory networks. In this study, we aimed to construct potential networks of bladder-cancer-related miRNAs and their known target genes using miRNA expression profiling and bioinformatics tools and to investigate potential key molecules that might play roles in bladder cancer regulatory networks. Global miRNA expression profiles were obtained using microarray followed by RT-qPCR validation using two randomly selected miRNAs. Known targets of deregulated miRNAs were utilized using DIANA-TarBase database v6.0. The incorporation of deregulated miRNAs and target genes into KEGG pathways were utilized using DIANA-mirPath software. To construct potential miRNA regulatory networks, the overlapping parts of three selected KEGG pathways were visualized by Cytoscape software. We finally gained 19 deregulated miRNAs, including 5 ups- and 14 down regulated in 27 bladder-cancer tissue samples and 8 normal urothelial tissue samples. The enrichment results of deregulated miRNAs and known target genes showed that most pathways were related to cancer or cell signaling pathways. We determined the hub CDK6, BCL2, E2F3, PTEN, MYC, RB, and ERBB3 target genes and hub hsa-let-7c, hsa-miR-195-5p, hsa-miR-141-3p, hsa-miR-26a-5p, hsa-miR-23b-3p, and hsa-miR-125b-5p miRNAs of the constructed networks. These findings provide new insights into the bladder cancer regulatory networks and give us a hypothesis that hsa-let-7c, hsa-miR-195-5p, and hsa-miR-125b-5p, along with CDK4 and CDK6 genes might exist in the same bladder cancer pathway. Particularly, hub miRNAs and genes might be potential biomarkers for bladder cancer clinics.

Is there a genetic predisposition for Turkish patients with sarcoidosis in the 329-bp region containing the BTNL2 rs2076530 polymorphism?

BACKGROUND/AIM: Sarcoidosis is a complex, multifactorial immune disorder with unknown etiology. A single nucleotide polymorphism (G→A, rs2076530) in the butyrophilin-like 2 (BTNL2) gene results in a truncating protein formation. It has been previously reported that this variation may be a risk factor for sarcoidosis in certain ethnic groups. This study was conducted to determine whether there is any genetic predisposition for the BTNL2 rs2076530 polymorphism in the 329-bp region in Turkish patients with sarcoidosis. MATERIALS AND METHODS: DNA samples were obtained from volunteers including 53 Turkish patients with sarcoidosis and 52 healthy controls. Analysis of the 329-bp region was carried out by polymerase chain reaction and sequencing of genomic DNA. RESULTS: We did not find any genetic variation except the rs2076530 polymorphism in the 329-bp region. The AA genotype was associated with an increased risk of sarcoidosis in a recessive model [P = 0.027, OR 2.56 (95% CI 1.02-6.49)], but it did not include a risk for sarcoidosis in a dominant model (P = 0.885). CONCLUSION: Our results emphasize the recessive characteristic of the rs2076530 polymorphism in Turkish patients with sarcoidosis. The lack of any genetic variation except rs2076530 in the 329-bp region is another significant finding for Turkish patients.

Conventional and molecular cytogenetic analyses in Turkish patients with multiple myeloma.

OBJECTIVE: Multiple myeloma (MM) is characterized by the accumulation and proliferation of malignant plasma cells, secreting monoclonal immunoglobulins and genetic abnormalities in MM have implications for disease progression and survival. In the present study, we investigated the frequency of chromosomal abnormalities (CA) in Turkish patients with MM, using interphase FISH and CC and evaluated the relationship between the rearrangements detected, prognosis and stage of disease. MATERIAL AND METHODS: We performed conventional cytogenetic and FISH studies in 50 patients to detect chromosome anomalies associated with MM. FISH probes were used to detect 13q14, 13q34, 17p13 deletions, IGH rearrangements, and monosomy and/or trisomy of chromosomes 5, 9, and 15. RESULTS: CC studies could be performed in 32 of 50 cases and five patients (15.6%) showed chromosomal aberrations while 27 (84.3%) had normal karyotypes. By FISH, eighteen percent (9/50) of cases were found to be normal for all parameters evaluated. Eighty-two percent (41/50) of the patients were positive for at least one abnormality. Chromosome 13 anomalies were detected in 54% (27/50) of cases. The second most common aberration observed is chromosome 15 aberrations (50%). CONCLUSION: Median survival rate was shorter in patients with one of the abnormalities including chromosome 13 aberrations, IGH rearrangements or P53 deletions. Chromosome 15 aberrations were significantly higher in patients with stage III disease (p=0.02). We conclude that FISH studies should be performed in conjunction with conventional cytogenetic analysis for prognosis in multiple myeloma patients.

Performance of MLPA as a screening method for aneuploidy in uncultured amniocytes.

OBJECTIVE: To test whether the Multiplex Ligation-dependent Probe Amplification (MLPA) technique can be used as a screening test for rapid diagnosis of aneuploidies in uncultured amniocentesis. MATERIAL AND METHODS: In this prospective blind study, MLPA with chromosomes 13,18,21,X and Y specific probe mixes was performed in 500 amniotic fluid samples. Chromosome copy numbers were determined by analyzing size and peak area for each MLPA probe. Results were compared with those of karyotyping/FISH. RESULTS: Conclusive test results were obtained in 98% of the samples, whereas 10 were inconclusive. In all conclusive tests, the MLPA results were concordant with that of cytogenetic and/or FISH analyses. There were no false-positive results. A case with 69,XXX triploidy could not be diagnosed by MLPA. In total, 28 aneuploidies were diagnosed. There were no false-positive results. The performance of each probe was determined. CONCLUSION: MLPA is a rapid, simple and reliable assay for aneuploidy screening in uncultured amniocytes.

The analysis of the relationship between A1298C and C677T polymorphisms of the MTHFR gene with prostate cancer in Eskisehir population.

Prostate cancer is the most common cause of cancer deaths in men and is a major health problem worldwide. Methylene tetrahydrofolate reductase (MTHFR) plays an important role for folate metabolism and is also an important source for DNA methylation and DNA synthesis (nucleotide synthesis). The objective of this study was to investigate the relationship between the A1298C and C677T polymorphisms of the MTHFR gene and prostate cancer in the Turkish population. In our study, 93 prostate cancer patients between the ages of 50-89 and a control group of 166 benign prostate hyperplasia patients were evaluated. C677T and A1298C polymorphism ratios were compared among these two groups, and an analysis was made to see if there is a statistically meaningful difference. In this study, it has been observed that C677T polymorphism of the MTHFR gene produces no statistically significant difference for T allele frequency and the genotype frequency in prostate cancer patients and male controls with benign prostate hyperplasia not having prostate cancer, whereas it has been observed that A1298C polymorphism produces a statistically significant difference for C allele frequency in prostate cancer patients and controls and that it also produces a statistically marginal significance for genotype frequencies.

Detection of deletions and/or amplifications of genes related with lung cancer by multiplex ligation-dependent probe amplification (MLPA) technique.

BACKGROUND: Lung cancer has the leading mortality rate among all cancers and it is the second most common cause of death following cardiovascular diseases.The aim of the study was determining deleted and/or amplified regions of 64 different loci previously associated with lung cancer, by using Multiplex Ligation-dependent Probe Amplification (MLPA). RESULTS: The most frequently seen deletions in lung cancerous tissues were in 2p, 3p, 13q, 17p, 16p and the most frequently seen amplifications were in 17q, 8p and 5q. We observed same deletions in the same regions in normal lung tissues as in cancerous tissues in lower frequencies. Deletions in 5q, 8p, 9q, 10p, 11p. 11q, 12p, 14q, 17q and 21q probe regions were seen especially in cancerous tissues. MATERIALS/METHODS: One hundred non small cell lung cancer (NSCLC) tissue samples which had been previously examined histopathologically were included in this investigation. DNA extracts of normal lung tissues from the same patients were used as control group in the study. CONCLUSIONS: As a conclusion, it was determined that MLPA is an alternative technique which can give cheap, fast and reliable results in the screening of lung cancers. The findings obtained in the study are compatible with the literature. MLPA is one of the most important molecular techniques which have been developed recently and it can be used in cancer screening easily and reliably.

Prognostic impact of chromosome alterations detected by FISH in Turkish patients with B-cell chronic lymphocytic leukemia.

The goal of this study was to evaluate the relation of chromosomal abnormalities detected by fluorescence in situ hybridization (FISH) in the prognosis of B-cell chronic lymphocytic leukemia (B-CLL) patients. We evaluated the common recurrent chromosomal aberrations in 79 B-CLL patients (51 men, 28 women; mean age 64.3+/-1.2) by FISH analysis using 11q22.3 (ATM), 13q14.3 (13S319 and 13S25), CEP12, and 17p13.1 (TP53) specific probes. Of the 79 patients analyzed by FISH, 40 or 50.6% had at least one aberration. In particular, 34 (43%) patients had a single abnormality and 6 (7.6%) patients had 2 abnormalities. The most frequent abnormality was 13q14.3 deletion, which was detected in 26 (32.9%) patients. Trisomy 12 was seen in 12 (15.2%) cases, and was followed by 17p13.1 (TP53) deletions and 11q22.3 (ATM) deletions in 6 (7.6%) and 4 (5.1%) patients, respectively. When the overall frequencies of these chromosomal aberrations were distributed according to RAI stages, the majority of patients with 13q14.3 deletion (55%), trisomy 12 (70%), and ATM or TP53 deletions (66.7 %) were in advanced stages of disease (RAI II-IV). The overall survival durations in good, intermediate, and poor prognostic groups were 51+/-1.3, 50.9+/-8.6, and 12+/-3.3 months, respectively. Our data suggests that FISH analysis of B-CLL patients provides important diagnostic, clinical, and prognostic information which may help clinicians assess the prognosis and make appropriate treatment decisions.

Genomic alterations in low-grade, anaplastic astrocytomas and glioblastomas.

To extend our understanding of potential stepwise genetic alterations that may underlie tumor progression from low-grade astrocytomas to glioblastomas, histopathologic and comparative genomic hybridization analyses were performed on tumor specimens from 68 primary lesions, including 40 glioblastomas, 10 anaplastic and 18 low-grade astrocytomas. The number of aberrations per case increased towards the higher grade tumors (grade II: 1.66+/-1.49; grade III: 2.80+/-1.68; grade IV: 3.02+/-1.07; F=6.955, p=0.002). A gain of 7/7q was common and the most frequently seen aberration in low-grade astrocytomas, whereas loss of 10q was the most frequently seen anomaly in anaplastic astrocytomas and glioblastomas. Chromosome 7p amplification was only detected in glioblastomas. Chromosome 10/10q deletion and combination of 1p, 19q and 17p deletions were specific to high-grade astrocytic tumors. Sequences of chromosome 7 and 10 seem to have pivotal roles in the biology of human gliomas. The genomic copy deletions of chromosomes 1p and 19q might provide an alternative mechanism in the genesis of astrocytomas.

Fish detected p53 deletion and N-MYC amplification in colorectal cancer.

BACKGROUND/AIMS: Data about p53 condition in sporadic colorectal cancer and its impact on clinical features is controversial, and studies regarding N-myc gene in colorectal cancer are limited in number. Our aim was to determine the frequency of p53 deletion and N-myc amplification by fluorescence in situ hybridization method in colorectal cancer and their relationship with clinical parameters. METHODOLOGY: The study was prospectively derived from 40 patients who were diagnosed as having colorectal cancer (Dukes' stages: 11 B, 18 C, 11 D) and went to surgery. Fresh tumor samples from all patients and adjacent normal tissue from 16 patients as control specimens were obtained. Locus specific fluorescence in situ hybridization probes was used for p53 and N-myc gene. For each sample, hundred interphase nuclei were analyzed based on their fluorescence phenotype. RESULTS: Both p53 allelic loss and N-myc amplification were found in 21 cancer tissues, while p53 allelic loss was not observed in any of control tissue, and N-myc amplification only in 1 (p<0.001, p<0.01). Mutations were significantly related with metastasis. CONCLUSIONS: Our study indicates that p53 and N-myc mutations are frequently found in tumor tissue of colorectal cancer and both gene alterations are correlated with the more aggressive tumor phenotype.