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Öğe 17p Deletion is associated with resistance of B-cell chronic lymphocytic leukemia cells to in vitro fludarabine-induced apoptosis(Taylor & Francis Ltd, 2007) Turgut, Burhan; Vural, Ozden; Pala, Funda S.; Pamuk, Gulsum E.; Tabakcioglu, Kiymet; Demir, Muzaffer; Ongoren, SenizWe explored the relationship between the cytogenetic/biologic characteristics of B-chronic lymphocytic leukemia (B-CLL) cells and their tendency to undergo spontaneous or fludarabine-induced apoptosis in vitro. B cells from 36 B-CLL patients were incubated with or without fluclarabine for 48 h. Apoptosis was determined by two assays: annexin V staining and DNA staining. Fluorescence in situ hybridization was used for detection of trisomy 12, 11q deletion, and 17p deletion. Bcl-2 and CD38 expressions were determined by flow cytometry. Five patients had 17p deletion, 6 had trisomy 12, and another 6 had 11q deletion. B-CLL cells with 17p deletion had significant resistance to apoptosis induced by fludarabine and a slight spontaneous resistance to apoptosis. Bcl-2 and CD38 were not associated with in vitro spontaneous and fludarabine-induced apoptosis. In conclusion, 17p deletion, which causes loss of p53 gene, is associated with resistance to fludarabine-induced apoptosis in vitro. New treatment modalities should be tried in B-CLL patients with 17p deletion.Öğe A case of Infective Endocarditis Caused by Pseudoclavibacter species(Ankara Microbiology Soc, 2022) Eryildiz, Canan; Mert, Haibe Tulin Elmaslar; Tabakcioglu, KiymetInfective endocarditis is an infectious disease usually caused by bacteria, including streptococci, staphylococci, and enterococci. In this report, a case of infective endocarditis in which Pseudoclavibacter spp. detected as the causative agent was presented. A 66-year-old female patient was admitted to our hospital with weight loss, malaise, and lumbar pain. A 2/6 murmur was detected in the physical examination of the patient, who had a history of mitral valve surgery nine years earlier. Blood sample was collected for culture and vancomycin [1 g/24 hours, intravenously (IV)] and gentamicin (80 mg/8 hours, IV) therapy were started. Gram-positive bacilli were detected on the second day of incubation in the blood culture bottle incubated in the BacT/ALERT 3D microbial detection system (bioMerieux, France). High uptake in the focal area of the mitral valve on positron emission tomography-computerized tomography (PET-CT) was interpreted as infective endocarditis. The patient was considered to be at very high risk for surgery and she was discharged after the vancomycin treatment completed for 42 days. For bacterial identification, DNA was isolated using a commercial kit (Invitrogen PureLink Genomic DNA Mini Kit; ThermoFisher Scientific, Waltham, MA, USA), and the 16S rRNA gene was amplified by a polymerase chain reaction (PCR) assay using universal primers 27F and 1492R. Sequence analysis of the PCR product was carried out on an Applied Biosystems 3730XL DNA Analyzer with an Applied Biosystems BigDye Terminator v3.1 Cycle Sequencing Kit (ThermoFisher Scientific, Waltham, MA, USA). A 1366 bp long sequence of the isolate was analyzed using the Basic Local Alignment Search Tool (BLAST) in GenBank and the sequence was 99% compatible with Gulosibacter spp. (GenBank sequence ID: LR884222.1, identity 1365/1366 bp) and Pseudoclavibacter spp. (GenBank sequence ID: H375951.1, identity 1364/1366 bp). Upon the negative result of nitrate reduction test of bacterium that was oxidasepositive, the bacterium was considered as Pseudoclavibacter spp. Linezolid, clindamycin, and tetracycline susceptibilities were determined by using the disc diffusion method. The isolate was susceptible to linezolid and resistant to clindamycin and tetracycline. It was also susceptible to penicillin [minimum inhibitory concentration (MIC) = 0.002 mu g/ml], vancomycin (MIC= 0.25 mu g/ml), and rifampicin (MIC= 0.003 mu g/ml) and was categorized as susceptible, increased exposure to ciprofloxacin (MIC= 0.25 mu g/ml), as determined by using a gradient strip test. Pseudoclavibacter species are non-spore-forming, non-motile, catalase-positive, aerobic gram-positive bacilli belonging to the Microbacteriaceae family. A limited number of clinical cases due to Pseudoclavibacter species have been reported. In the present case, gram-positive bacilli were considered to be the causative agent of infective endocarditis because of the growth of the same bacteria in six bottles of blood cultures taken at different times and increased focal involvement on PET-CT. To our knowledge, this is the second reported case of infective endocarditis due to Pseudoclavibacter species. In conclusion, in cases of clinical compliance in patients with prosthetic heart valves, gram-positive bacilli should be considered causative agents of infective endocarditis and in such cases, a range of microbiological assays should be used for identification.Öğe Effect of lipoic acid on paraoxonase-1 and paraoxonase-3 protein levels, mRNA expression and arylesterase activity in liver hepatoma cells(General Physiol And Biophysics, 2017) Ozgun, Eray; Ozgun, Gulben Sayilan; Tabakcioglu, Kiymet; Gokmen, Selma Suer; Sut, Necdet; Eskiocak, SevgiParaoxonase-1 (PON1) and paraoxonase-3 (PON3) are anti-atherosclerotic enzymes, synthesized primarily in liver and bound to HDL in circulation. The aim of the present study was to investigate the effects of therapeutic doses of lipoic acid on PON1 and PON3 protein levels, mRNA expression and arylesterase activity in liver. We treated HepG2 cells with 10, 40 and 200 mu M lipoic acid for 72 h. Cell viability was evaluated by 3-(4,5-dimethy1-2-thiazoly1)-2,5-dipheny1-2Htetrazolium bromide assay. PON1 and PON3 protein levels were measured by Western blotting, their mRNA expression was measured by quantitative PCR and arylesterase activity was measured spectrophotometrically. 200 mu M lipoic acid caused a significant increase on PON1 and PON3 protein levels and arylesterase activity as compared with control, 10 mu M and 40 mu M lipoic acid treated cells. 200 mu M lipoic acid also caused a significant decrease on PON1 mRNA expression whereas on a significant increase PON3 mRNA expression as compared with control, 10 mu M and 40 mu M lipoic acid-treated cells. Our study showed that although lip oic acid up-regulates PON3 but down-regulates PON1 mRNA expression, it increases both PON1 and PON3 protein levels and arylesterase activity in HepG2 cells. We can report that lipoic acid may be useful for preventing atherosclerosis at therapeutic doses.Öğe The Effects of Micronuclei with Whole Chromosome on Biological Dose Estimation(Tubitak Scientific & Technological Research Council Turkey, 2008) Pala, Funda S.; Alkaya, Fadime; Tabakcioglu, Kiymet; Tokatli, Fuesun; Uzal, Cem; Parlar, Sule; Alguenes, CetinThe total micronucleus (MN) assay has been used for purposes of biological dosimetry for many years. The variable spontaneous incidence of micronuclei in peripheral blood lymphocytes affects the sensitivity of biological dose estimations at low doses. It has been suggested that this problem could be solved by using the micronuclei-centromere assay. In this study. Co-60 gamma ray dose response curves for micronuclei (MN) and micronuclei without centromeres (MNC-) in the range of 0-5.0 Gy were established using a pancentromeric FISH probe on cultured binucleate lymphocytes from 2 donors. There were no significant inter-donor differences in the dose responses for either MN or MNC-. The relative proportions of MN that contained centromeres (MNC+) decreased with radiation dose, which is in line with the proposition that radiation predominately causes chromosomal breakage rather than whole chromosome loss. The a coefficients of MNC- curves decreased to 62% of the values for total MN whilst the beta coefficients were unchanged. MN and MNC+ frequencies of 60 control smoker and 40 non-smoker donors were also compared. No effect of smoking was observed. However the MNC+ spontaneous frequencies showed an age and gender effect with the highest frequencies in older women.Öğe Effects of permissible maximum-contamination levels of VOC mixture in water on total DNA, antioxidant gene expression, and sequences of ribosomal DNA of Drosophila melanogaster(Springer Heidelberg, 2015) Doganlar, Oguzhan; Doganlar, Zeynep Banu; Tabakcioglu, KiymetIn this study, we aimed to investigate the mutagenic and carcinogenic potential of a volatile organic compound (VOC) mixture with references to the response of D. melanogaster using selected antioxidant gene expressions, RAPD assay and base-pair change of ribosomal 18S, and the internal transcribed spacer, ITS2 rDNA gene sequences. For this purpose, Drosophila melanogaster Oregon R, reared under controlled conditions on artificial diets, were treated with the mixture of thirteen VOCs, which are commonly found in water in concentrations of 10, 20, 50, and 75 ppb for 1 and 5 days. In the random amplified polymorphic DNA (RAPD) assay, band changes were clearly detected, especially at the 50 and 75 ppb exposure levels, for both treatment periods, and the band profiles exhibited clear differences between the treated and untreated flies with changes in band intensity and the loss/appearance of bands. Quantitative real-time PCR (qRT-PCR) analysis of Mn-superoxide dismutase (Mn-SOD), catalase (CAT) and glutathione-synthetase (GS) expressions demonstrated that these markers responded significantly to VOC-induced oxidative stress. Whilst CAT gene expressions increased linearly with increasing concentrations of VOCs and treatment times, the 50- and 75-ppb treatments caused decreases in GS expressions compared to the control at 5 days. Treatment with VOCs at both exposure times, especially in high doses, caused gene mutation of the 18S and the ITS2 ribosomal DNA. According to this research, we thought that the permissible maximum-contamination level of VOCs can cause genotoxic effect especially when mixed.Öğe Evaluation of Frequencies of HLA-A, B and DR in Thracian Population and Examination of its Relationship With Balkan Populations(Aves Yayincilik, Ibrahim Kara, 2008) Pala, Funda Sibel; Tabakcioglu, Kiymet; Alguenes, Cetin; Oemuerlue, Imran KurtObjectives: In this study, human leukocyte antigen (HLA) allel frequencies of Thracian Turkish population were determined. Study Design: The study group consisted of 105 tissue donors who live in Thrace region of Turkey for three generations and have similar linguistic features. Polymerase chain reaction-sequence-specific primer (PCR-SSP) method was used for genotyping of HLA-A, B and DRB1 alleles. The most frequent HLA alleles were HLA-A*02 (20.5%), HLA-B*35 (22.9%) and HLA-DR*11 (17.6%). Results: Frequencies of HLA alleles show some variations among different populations because of their highly polymorphic gene structure. Determining the distribution of HLA alleles is one of the most preferred genetical approaches for clarifying the relationship between populations. Conclusion: In this study also, HLA-DR allel frequencies were compared in order to determine the relationship between other Turkish populations and Balkan populations. It is observed that Thracian Turkish population has similar HLA-DR distributions with Balkan populations.Öğe An Evaluation of the Effect of the Biological Dose of Fluoroscopic Radiation Exposure in the Operating Room(Aves, 2018) Yilmaz, Baris; Copuroglu, Cem; Tabakcioglu, Kiymet; Pala, Funda Sibel; Ozcan, Mert; Ciftdemir, MertObjective: Through an evaluation of the biological dose, we aimed to evaluate the risks of ionizing radiation to which physicians and auxiliary healthcare personnel working in orthopedic operating rooms are exposed to via diagnostic use of fluoroscopy. Methods: Blood samples were collected from physicians and auxiliary healthcare personnel working in the orthopedic operating room. The biological dose was evaluated using micronucleus and dicentric analysis. To assess the effects of physical and chemical agents together, a total of 31,000 binucleate cells were evaluated using the micronucleus method and 16,500 metaphase plaques were evaluated using dicentric analysis, which is accepted as the most important indicator in determining the effects of radiation. Results: The study participants comprised 18 males and 5 females (16 physicians, 4 nurses, and 3 patient carers) with a mean age of 34.1 years (range, 22-58 years) who were thought to have been exposed to ionizing radiation in the working environment. The mean duration of working under ionizing radiation was 73.6 months (range, 1.5-420 months). In the blood samples, the total micronucleus frequency was determined as 8.8 +/- 1.4. In the evaluation of the 16,500 metaphysis plaques, radiation-specific dicentric was observed in 5 subjects (normal frequency: 5/10,000). As a result of the analysis made use both methods, the dose was determined to be slightly above background level, and below risk level in 6 subjects. The dose was related with medical applications in 4 of these subjects. Conclusion: Fluoroscopy should be attempted in the operating room within a restricted time as far as possible and at measurements of high kV and low mA. kV-mA values are of utmost importance for providing the best results according to the nature of the operation; the tube outlet is predefined away from the patient's skin.Öğe Genotoxic Effects of Heavy Metal Mixture in Drosophila melanogaster: Expressions of Heat Shock Proteins, RAPD Profiles and Mitochondrial DNA Sequence(Springer International Publishing Ag, 2014) Doganlar, Zeynep Banu; Doganlar, Oguzhan; Tabakcioglu, KiymetThe genotoxic effects of four heavy metal mixtures on Drosophila melanogaster were investigated with reference to gene expressions of heat shock proteins (HSP26, HSP60, HSP70 and HSP83), DNA profiles, and mitochondrial NADH dehydrogenase sequence. Adult D. melanogaster flies were treated with a mixture of four (Fe, Cu, Cd and Pb) heavy metals (HMs) in three different concentrations, which were selected based on one higher dose (HM3) and one lower dose (HM1) relative to the permitted limits (HM2) in drinking water at 1st, 5th and 10th days. It was determined that the amount of the accumulated heavy metals and the expressions of the HSP genes were changed with increasing exposure time. The accumulations of Cd and Pb were increased with increasing exposure time; additionally, the HSP expression patterns were determined as HSP70>HSP60>HSP26>HSP83 HM1 (5th day), HM2 (5th day and 10th day), and HM3 (all exposure times). It was also determined that the application of the heavy metal mixture affected the random amplified polymorphic DNA (RAPD) profiles and the mitochondrial NADH dehydrogenase sequence of D. melanogaster. The highest base pair changes (9 bp) were determined at the HM2 concentration (permissible limits in drinking water) on the 1st day of treatment. Therefore, it was shown that mixture of four heavy metals caused a genotoxic effect and D. melanogaster is a useful model organism for heavy metal-induced genotoxicity studies.Öğe Genotyping and Identification of Antigen B Gene Polymorphism of Echinococcus granulosus in Edirne, Thrace, and the First Report of Genotype G2 (Tasmanian Sheep Strain) in Turkey(Galenos Publ House, 2022) Tarladacalisir, Taner; Eryildiz, Canan; Tabakcioglu, Kiymet; Sakru, NerminBackground: Echinococcus granulosus is the causative agent of cystic echinococcosis in humans and livestock. It is common worldwide. Cystic echinococcosis is still an important public health problem in Turkey, which is an endemic region.Aims: To genotype Echinococcus granulosus isolates and investigate antigen B gene polymorphism in Thrace, Turkey.Study Design: A cross-sectional study.Methods: Seventy-five hydatid cyst materials obtained between June 2020 and May 2021 were included in the study. Hydatid cyst materials were collected from 12 humans from various hospitals in Edirne and 63 from slaughterhouse animals during the same period. Cyst materials were localized in 8 livers and 4 lungs in humans, 23 livers and 17 lungs in cattle, and 13 livers and 10 lungs in sheep. In the first step, the 12S ribosomal RNA gene was amplified by polymerase chain reaction for all samples and run on an agarose gel. Band patterns were used for strain typing. Then, the selected samples that represented each of the band patterns obtained by single-strand conformation polymorphism analysis were sequenced for AgB1, AgB2, mt-CO1, and mt-ND1 genes.Results: Three different genotypes in Edirne, Thrace, Turkey, were observed for Echinococcus granulosus: G1 (domestic sheep strain), G2 (Tasmanian sheep strain), and G3 (buffalo strain). G1 was the dominant genotype in Edirne, and G3 was the second most common. Additionally, polymorphism in AgB1 and AgB2 gene regions was found.Conclusion: This study is the first to report on Echinococcus granulosus G2 (Tasmania sheep strain) in Turkey and G3 (buffalo strain) and antigen B polymorphism in Thrace. The study results will contribute to the prevention and control programs for cystic echinococcosis in Turkey and worldwide.Öğe Investigation of antimicrobial resistance and virulence genes of Campylobacter isolates from patients in a tertiary hospital in Edirne, Turkey(Elsevier, 2020) Eryildiz, Canan; Tabakcioglu, Kiymet; Kuyucuklu, Gulcan; Sakru, NerminPurpose: Campylobacter is one of the most common pathogens that cause food-borne infections worldwide. The aim of this study was to determine the antimicrobial resistance rates and the presence of multiple virulence genes in Campylobacter isolates obtained from humans. Materials and Methods: In this study, 71 Campylobacter isolates obtained from human faecal samples were used. Antimicrobial susceptibility tests were performed through the gradient strip method. The presence of virulence genes was investigated by monoplex and multiplex polymerase chain reaction. Results: The rate of resistance of the 66 Campylobacter jejuni isolates was 12.1% for erythromycin, 40.9% for tetracycline and 68.2% for ciprofloxacin. Only one of five Campylobacter coli isolates was resistant to these three antimicrobial agents. The flaB, pldA, cdtA, cadF, cdtC and ceuE genes were found in all 66 of the C. jejuni isolates. In the C. jejuni isolates, positivity rates of 92.4% for flaA, 96.7% for cdtB, 98.5% for ciaB, 90.9% for dnaJ and 96.7% for racR were observed. The flaA, flaB, ciaB, cdtA and cdtC genes were present in all C. coli isolates. Conclusions: It was detected that there is an increase in antimicrobial resistance of Campylobacter strains in our region, and most of the isolates harbour virulence genes.Öğe Lysinibacillus massiliensis Isolated from the Synovial Fluid: A Case Report(Bilimsel Tip Yayinevi, 2020) Eryildiz, Canan; Tabakcioglu, Kiymet; Kehaya, Sezgin; Sakru, Nermin; Gurcan, SabanLysinibacillus massiliensis is an aerobic, endospore-forming, gram-negative staining bacterium with peritrichous flagella belonging to the Bacillaceae family. A few cases of L. massiliensis isolated from the cerebrospinal fluid and tissue have been reported. In this study, we aimed to describe a case of L. massiliensis isolated from the synovial fluid. The synovial fluid from a 74-year-old female patient was inoculated into blood culture bottle. Gram-negative rods were observed in a gram-stained smear from a positive blood culture bottle. The bacterium was identified as Lysinibacillus sphaerkus/Lysinibacillus fusiformis, with a probability of 89% using an automated bacterial identification system (VITEK2; Biomerieux, France). Subsequently, 16S rRNA gene sequencing was performed, and the sequence was analyzed using the Basic Local Alignment Search Tool. The sequence had 99.9% (1426/1427) identity with the strain L. massiliensis (GenBank ID: NR_043092.1). To our knowledge, this is the first reported case of L. massiliensis isolated from the synovial fluid. When an endospore-forming gram-negative staining bacterium can not be identified by phenotypic characterization, L. massiliensis should be considered, and different microbiological methods should be used for identification.Öğe MEFV Gene Exon 2 and Exon 10 Gene Region Mutations of Familial Mediterranean Fever Patients in Trakya Population(Aves Yayincilik, Ibrahim Kara, 2010) Gurkan, Hakan; Ozkayin, Emine Nese; Tabakcioglu, Kiymet; Algunes, CetinObjectives: The objective of the study is to explore the MEFV gene exon 2 and exon 10 gene region mutations which take place in etiopathogenesis of Familial Mediterranean Fever in the Thrace population with the DNA sequence analysis method and to compare the results with the other studies. Patients and Methods: The study included patients with Familial Mediterranean Fever who have no relative relationship, have the same linguistic characteristic and live in the Thrace region for at least three generations (34 females, 34 males). MEFV gene exon 2 and exon 10 gene regions multiplied with PCR and their nucleotids were determined with the DNA sequence analysis method. Results: G442C, T306C, A414G, C495A, G605A SNPs were found in MEFV gene exon 2 gene region and G20400, A2080G, 02082A, A2084G, T2177C, G2282A SNPs were found in MEFV gene exon 10 gene region in the Thrace population. Conclusion: The T306C, A414G, C495A, G605 single-nucleotide polymorphisms in MEFV gene exon 2 gene region and the mutations in exon 10 gene region are not compatible in terms of their frequencies with the results of the other studies.Öğe Molecular Identification of Campylobacter Species Isolated from Patients with Gastroenteritis in Edirne, Turkey(Galenos Publ House, 2022) Eryildiz, Canan; Sakru, Nermin; Tabakcioglu, Kiymet; Ugur, Mediha Cerrah; Bukavaz, SebnemBACKGROUND/AIMS: Campylobacter is a major cause of foodborne diarrheal disease, and the incidence of campylobacteriosis has significantly increased in both developed and developing countries. The purpose of the present study was to identify the species of Campylobacter isolates and to evaluate the distribution of Campylobacter infections according to various characteristics in our region. MATERIALS AND METHODS: Campylobacter isolates obtained from patients at a tertiary hospital in Edirne, Turkey were included in this study. The distribution of Campylobacter infections was evaluated according to age, season, and gender. Species identification was performed by multiplex polymerase chain reaction (PCR). The RNA polymerase beta-subunit gene (rpoB) of selected samples was amplified, and DNA sequencing was performed. RESULTS: Campylobacter species were isolated from 226 (4.3%) of the 5,241 samples. One hundred and seventy-six (89.3%) of 197 samples were identified as C. jejuni and 19 (9.6%) as C. coli by multiplex PCR. Two isolates showed a band profile compatible with both C. jejuni and C. coli. DNA sequencing was performed for 21 isolates. Sixteen isolates were compatible with C. jejuni and 5 isolates were consistent with C. coli. There was no statistically significant difference in Campylobacter isolation rates according to gender and season (p>0.05). Campylobacter species were most frequently isolated from children in the age group of 0-14 years (p<0.01). CONCLUSION: Campylobacter is one of the main causes of diarrhea in Turkey, and this infection is more common in children. This study contributes to information about the situation of Campylobacter infection and the genetic features of isolates in Turkey.
