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Öğe Aqueous humor penetration of topically applied ciprofloxacin, ofloxacin and tobramycin(Ecv-Editio Cantor Verlag Medizin Naturwissenschaften, 1997) Durmaz, B; Marol, S; Durmaz, R; Oram, O; Hepsen, IF; Gunal, SThe purpose of this study was to determine the aqueous humor concentrations of topically applied ciprofloxacin (GAS 86393-32-0), ofloxacin (GAS 82419-36-1) and tobramycin (GAS 79645-27-5). Thirty patients undergoing cataract extraction or trabeculectomy were randomly divided into three groups and each of the group received either 0.3 % ciprofloxacin, ofloxacin or tobramycin topical drops preoperatively. Eyedrops were instilled for six times at a frequency of one drop every 15 minutes, beginning 90 minutes before initiation of the surgery. At the time of surgery, 0.1 mi aqueous fluid was aspirated from the anterior chamber. Concentrations of the antimicrobial agents were determined using the microbroth dilution procedure outlined by the National Committee for Clinical Laboratory Standards. Escherichia coli (ATCC 25922) was used as a standard strain for determination of minimal inhibitory concentrations (MICs). The mean aqueous levels of ciprofloxacin and ofloxacin were found to be 0.092 +/- 0.077 mu g/ml, 0.964 +/- 0.693 mu g/ml, respectively. Tobramycin did not reach the concentration that could be detected by applied method. Conclusion: The mean aqueous humor levels of ofloxacin and ciprofloxacin were more than the MICs levels for most of the ocular pathogens which may cause postoperative endophthalmitis.Öğe Genotyping of Mycobacterium tuberculosis clinical isolates in two cities of Turkey(Bmc, 2005) Zozio, T; Allix, C; Gunal, S; Saribas, Z; Alp, A; Durmaz, R; Fauville-Dufaux, MBackground: Population-based bacterial genetics using repeated DNA loci is an efficient approach to study the biodiversity and phylogeographical structure of human pathogens, such as Mycobacterium tuberculosis, the agent of tuberculosis. Indeed large genetic diversity databases are available for this pathogen and are regularly updated. No population-based polymorphism data were yet available for M. tuberculosis in Turkey, at the crossroads of Eurasia. Results: A total of 245 DNAs from Mycobacterium tuberculosis clinical isolates from tuberculosis patients residing in Turkey (Malatya n = 147 or Ankara n = 98) were genotyped by spoligotyping, a high-throughput genotyping method based on the polymorphism of the Direct Repeat locus. Thirty-three spoligotyping-defined clusters including 206 patients and 39 unique patterns were found. The ST41 cluster, as designated according to the international SpolDB3 database project, represented one fourth and when gathered to three genotypes, ST53, ST50 and ST284, one half of all the isolates. Out of 34 clinical isolates harboring ST41 which were further genotyped by IS6110 and by MIRU-VNTR typing, a typical 2-copy IS6110-RFLP pattern and a 215125113322 MIRU-VNTR pattern were observed among 21 clinical isolates. Further search in various databases confirms the likely Turkish-phylogeographical specificity of this clonal complex. Conclusion: We described a new phylogeographically-specific clone of M. tuberculosis, designated LAM7-TUR. Further investigations to assess its frequency within all regions of Turkey and its phylogeographical origin and phylogenetic tree will shed new light on its endemicity in Asia Minor.Öğe Genotyping of rifampin-resistant Mycobacterium tuberculosis isolates from western Turkey(K Faisal Spec Hosp Res Centre, 2004) Cavusoglu, C; Durmaz, R; Bilgic, A; Gunal, SBackground: Although the rate of multiple drug resistance is high, there is no published data on the transmission rate of drug-resistant strains of Mycobacterium tuberculosis in the Aegean region of western Turkey that are based on molecular methods. Methods: IS6110 and pTBN12 restriction fragment length polymorphism (RFLP) methods were used for typing M. tuberculosis strains isolated from 26 sputum samples from 26 patients. Results: Nineteen of the rifampin-resistant isolates (73.1%) contained 6 to 11 copies of IS6110. Eighteen different IS6110 DNA fingerprint patterns were observed in the 26 rifampin-resistant isolates. Twenty-three of the 26 rifampin-resistant isolates were also resistant to isoniazid. When evaluated together, both methods yielded 21 (80.9%) different banding patterns and the level of clustering was 34.6%. The average number per pattern was 1.23 (26/21). Conclusions: IS6110 fingerprinting suggests that the rifampin-resistant isolates obtained from the Aegean region had a relatively high clustering rate and were clonally related. These findings showed that the rifampin-resistant isolates are actively transmitted between patients. Urgent measures should be taken to prevent the spread of these resistant strains.Öğe The level of endothelin-1 and nitric oxide in patients with chronic viral hepatitis B and C and correlation with histopathological grading and staging(Wiley, 2006) Ersoy, Y; Bayraktar, NM; Mizrak, B; Ozerol, IH; Gunal, S; Aladag, M; Bayindir, YBackground and aim: The aim of this study was to estimate the serum levels of endothelin-1 (ET-1) and nitric oxide (NO) and to analyze the correlation of their levels with histopathological grading and staging of the liver in patients with chronic hepatitis B (CHB) and C (CHC). Methods: Eighty-nine patients who were either HBsAg positive (45 CHB patients, 34 inactive carriers (IQ) or had CHC (10 patients) and 36 healthy volunteers as a control group were included in this study. Fifty patients from the CHB (n = 43) or CHC (n = 7) groups with elevated serum alanine transaminase (ALT) levels underwent a liver biopsy. Histological activity was scored according to Ishak's activity and the fibrotic index. The ET-1 serum concentration was determined with a commercially available ELISA assay kit. Total nitrite was measured by the Griess reaction as an index for NO production. Results: Serum levels of ET-1 and NO were significantly increased in CHB patients (7.67 +/- 4.00 pg/ml and 172.44 +/- 50.30 mu mol/l, respectively) compared with the IC group (3.99 +/- 5.42 pg/ml and 114.68 +/- 32.22 mu mol/l, respectively) and the control group (3.05 +/- 0.65 pg/ml and 58.61 +/- 24.18 mu mol/l, respectively) (p < 0,000 1). The CHC patients also had significantly higher serum levels of ET- 1 (5.92 +/- 4.24 pg/ml) and NO (147.50 +/- 55.84 mu mol/l) compared to the control group (p < 0.0001 and < 0.001, respectively). Linear regression analysis identified that the level of ET- I was an independent variable that correlated significantly with the stage score (r(2) = 0.348, p < 0.0001) in CHB patients but there was no correlation in the CHC group. Conclusion: ET-1 and NO levels were increased in chronic hepatitis and there was a significant correlation between the ET-I level and the stage in CHB patients. (c) 2005 Elsevier Ireland Ltd. All rights reserved.Öğe Molecular epidemiology of tuberculosis in Turkey(Blackwell Publishing Ltd, 2003) Durmaz, R; Gunal, S; Yang, Z; Ozerol, IH; Cave, MDOur objective was to determine the extent of fingerprint pattern diversity of Mycobacterium tuberculosis isolates from Turkey. Of the 320 patient isolates, 81 (25.3%) carried less than or equal to5 copies of IS6110. The combination of two typing procedures on isolates from 317 patients identified 157 strains as unique and clustered 160 isolates in 59 clusters. In spite of the fact that the patients originated from a large geographic area and represented only a small fraction (1.5%) of the patients in the country, 50.5% of the isolates were clustered.Öğe Primary drug resistance and molecular epidemiology of Mycobacterium tuberculosis isolates from patients in a population with high tuberculosis incidence in Turkey(Mary Ann Liebert, Inc, 2003) Durmaz, R; Ozerol, IH; Durmaz, B; Gunal, S; Senoglu, A; Evliyaoglu, ETo determine the rate of primary drug resistance and compare the fingerprint pattern diversity of the resistant and sensitive Mycobacterium tuberculosis isolates, antituberculosis susceptibility testing and restriction fragment length polymorphism (RFLP) analysis were performed on 88 M. tuberculosis isolates of the patients who were diagnosed as new tuberculosis cases in 2000. Primary resistance to isoniazid, rifampicin, ethambutol, and streptomycin were determined by the BACTEC method. IS6110 and pTBN12 were used as molecular markers. The frequency of resistance to at least one drug was 32.95%, whereas 10.23% of the isolates were resistant to more than one drug. Single-drug resistance to isoniazid, streptomycin, ethambutol, and rifampicin was found in 9 (10.22 %), 7 (7.95 %), 4 (4.54 %), and 0 (0.0 %) strains, respectively. Two M. tuberculosis strains (2.26%) showed multiple drug resistance. The combination of two fingerprinting procedures on a total of 88 isolates identified 58 (65.9%) strains as unique and clustered 30 strains in 11 clusters (clustering = 34.1%). The clustering rate for resistant and sensitive isolates was 13.8% and 40.1%, respectively. In conclusion; drug susceptibility testing showed that the majority of the drug-resistant infections involved either isoniazid or streptomycin alone. In addition to the high tuberculosis incidence, elevated primary drug resistance and high clustering rate indicate problems in the present control programs. New control strategies supported by molecular typing might be more effective to reduce tuberculosis.Öğe Sensitivity of two-stage PCR amplification for detection of Mycobacterium tuberculosis in paraffin-embedded tissues(Elsevier Science Bv, 1997) Durmaz, R; Aydin, A; Durmaz, B; Aydin, NE; Akbasak, BS; Gunal, SIn order to improve the sensitivity of polymerase chain reaction (PCR) for the detection of mycobacterial DNA in paraffin-embedded tissues, a new approach with two sets of specific primers in two-stage PCR was employed in specimens obtained from tuberculosis patients and controls. Thirty-nine paraffin blocks selected from patients who had been diagnosed as having tuberculosis by radiological evaluations, histopathological findings,land clinical symptoms and signs including response to antituberculous treatment were examined. The control group consisted of 10 specimens from individuals that were proved to be negative for tuberculosis. After deparaffinization, lysis, phenol-chloroform extraction, and ethanol precipitation, the isolated DNA was amplified by PCR. Initially, all specimens were examined by the one-stage PCR using specific primers for 123-base pair (bp) fragment in IS6110 of mycobacterial DNA which yielded positive results only in 3 out of 39 (7.7%). In the two-stage PCR technique, 245-bp fragment of mycobacterial DNA was amplified at the first-step, then the PCR products were reamplified using the second specific primer pairs for 123-bp fragment. The true positivity of the two-stage PCR was 84.6% (33/39). The results indicate that two-stage PCR is more sensitive than one-stage (84.6% vs. 7.7%). All control specimens were negative by both PCR amplification methods, indicating that specificity of both methods was high. When the two-stage amplification was used, PCR positivity in the specimens obtained from different tissues was as follows: peritoneal and omental biopsies, 4/4; bone biopsies, 3/3; lymph node biopsies, 12/14; genito-urinary biopsies, 7/9; skin biopsies, 4/6; and one from each lung, breast, and pleural biopsies. PCR showed a good correlation with the granulomatous tissue reaction resulting in a 83.8% (31/37) positivity. The results indicate that the two-stage PCR amplification can be used for detection of M. tuberculosis in paraffin-embedded tissues and is a useful technique in confirming tuberculosis in patients with clinically suspected disease who have acid-fast stain-negative. (C) 1997 Elsevier Science B.V.Öğe Simultaneous detection of isoniazid, rifampin, and ethambutol resistance of Mycobacterium tuberculosis by a single multiplex allele-specific polymerase chain reaction (PCR) assay(Elsevier Science Inc, 2005) Yang, ZH; Durmaz, R; Yang, D; Gunal, S; Zhang, LX; Foxman, B; Sanic, APrompt detection of drug resistance of Mycobacterium tuberculosis is essential for effective control of tuberculosis (TB). We developed a multiplex allele-specific polymerase chain reaction (MAS-PCR) that detects the most commonly observed isoniazid (M), rifampin (RIF), and ethambutol resistance-associated mutations in a single assay. The usefulness of the newly developed method was evaluated with 174 clinical isolates of M. tuberculosis obtained from Turkey. Distinct PCR banding patterns were observed for different mutation profiles and the correlation between MAS-PCR results and DNA sequencing findings was 99.4%. With culture-based phenotypic drug susceptibility testing as a reference standard, the sensitivity and specificity of the newly developed MAS-PCR assay for drug resistance-related genetic mutation detection were determined to be 81.1% and 97.5% for INH, 93.0% and 98.9% for RIF, and 54.5% and 68.0% for ethambutol. MAS-PCR provides a rapid, potentially more cost-effective, method of detecting multidrug-resistant TB. (c) 2005 Elsevier Inc. All rights reserved.
