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Öğe ANTIOXIDANT ACTIVITY AND FATTY ACID COMPOSITION OF GINKGO BILOBA FROM TURKEY(WILEY-BLACKWELL, 2011) Maltas, Esra; Vural, Hasibe Cingilli; Yildiz, SalihIn this study, we investigated the antioxidant capacity and total phenolic content of the extracts Ginkgo biloba from Turkey. The antioxidant activity of the methanolic and acetone extracts from G. biloba leaves was measured by various assays, including ferric reducing antioxidant power assay, cupric reducing antioxidant capacity assay and metal chelating capacity. Total phenolic content of the extracts was measured as gallic acid equivalents (GAE) by Folin-Ciocalteu reagent. The methanolic extract showed higher antioxidant activity related to high phenolic content with 76.0 +/- 5.2 mg GAE/g dry weight. Fatty acid compositions of the methanolic and acetone extracts of G. biloba were analyzed. Data suggested that G. biloba grown in Turkey may be an important source of natural antioxidant.Öğe Biochemical and molecular analysis of soybean seed from Turkey(ACADEMIC JOURNALS, 2011) Maltas, Esra; Dageri, Nazan; Vural, Hasibe Cingilli; Yildiz, SalihWe aimed molecular analysis of soybean by lecithin specific primer pairs LE5/LE6. Genomic DNA was extracted from soybean by CTAB method and EZ1 nucleic acid isolation system. A sensitive qualitative detection method for soybean, using the polymerase chain reaction was developed with E5/LE6 primers, produced a 195 bp product. However, the antioxidant activity of methanolic extract of soybean from Turkey by using DPPH and ABTS radical scavenging assays, showed higher antioxidant activity with 53.19 +/- 0.87% and 45.10 +/- 0.32%, respectively. Fatty acid compositions and several phenolic acids and flavonoids of soybean extract were analysed by gas and high performenace chromatography. Data showed that, the main compounds of the extract were eriodictyol, naringenin and linoleic acid.Öğe Design of a Probe Based on Poly(glycidyl methacrylate-co-vinylferrocene)-Coated Pt Electrode for Electrochemical Detection of PTEN Gene in PCR Amplified Samples from Prostate Tissues(WILEY, 2014) Bas, Salih Zeki; Maltas, Esra; Sennik, Busra; Yilmaz, Faruk; Vural, Hasibe CingilliIn this study, a new probe based on immobilization of amino linked oligonucleotide (NH2-linked DNA) on poly(glycidyl methacrylate-co-vinylferrocene)-coated Pt electrode was fabricated for the electrochemical detection of PTEN gene from human prostate tissues. The experimental parameters such as DNA immobilization time, DNA concentration, and target concentration were optimized. The selectivity of the NH2-linked DNA probe was assessed with mismatch (MM) and noncomplementary (NC) sequences. The applicability of the NH2-linked DNA probe to the PCR amplified samples correspond to PTEN gene from prostate tissues was evaluated. The immobilization of DNA on the copolymer was confirmed by FTIR, AFM, CV and DPV analysis. The PCR products were also identified by using agarose gel electrophoresis. The prepared probe indicated a linear range (10-100 g mL(-1)) with a detection limit (4.7 g mL(-1)) and a good selectivity of the NH2-linked DNA probe toward target DNA sequence. (c) 2014 Wiley Periodicals, Inc.Öğe An EPR Study On Cytosine Irradiated(WILEY, 2011) Usta, Ayhan; Vural, Hasibe Cingilli; Usta, Keziban; Aras, Erdal; Ceylan, Yusuf; Ozmen, AyhanIn this study, paramagnetic centers over the cytosine were formed by photolysis then these centers were investigated using EPR method. EPR signals were not recorded from non-irradiated the cytosine, but irradiated polycrystalline exhibited complex EPR spectra. For obtaining of cytosine polycrystalline, novel crystallization method was performed on powder cytosine. Effective crystallization conditions were achieved by adjustment of the concentration of the metal ions, chemical solutions, NaCl, KCl, glacial asetic acid, nitric oxide, percloric acid, glutamic acid, and pH of buffer. Cytosine (C4H5N2O) polycrystalline obtained were irradiated with Co-60 - rays at room temperature for 24 and 72 h. At the sample irradiated for 72 h, the paramagnetic centers were determined between 120 and 450 K by X-band EPR spectrometer. The spectra were found to be dependent slightly on temperature. Two cation radicals were determined in the structure and these were called Radicals I and II. The g and hyperfine constants were found to be a(H2a) = 61 G, a(N2) = 9.39 G, a(N1) = 7.15 G, and g(1) = 2.0026 for the Radical I; a(H3) = 10.57 G, a(H1) = 3 G, a(N3) = 6.72 G, a(N1) = 5.36 G for, and g(2) = 2.0034 the Radical II. Copyright (C) 2010 John Wiley & Sons, Ltd.Öğe Extraction of genomic DNA from polysaccharide- and phenolics-rich Ginkgo biloba(ACADEMIC JOURNALS, 2011) Maltas, Esra; Vural, Hasibe Cingilli; Yildiz, SalihOne prerequisite to reliable molecular biology work is that the genomic DNA of a sample should be of good quality. The isolation of intact, high-molecular-mass genomic DNA is essential for many molecular biology applications including long Polymerase Chain Reaction (PCR), endonuclease restriction digestion, Southern blot analysis, and genomic library construction. Many protocols are available for the extraction of DNA from plant material. DNA extraction of Ginkgo biloba is quite difficult to work on because of the high phenolic and polysaccharide content of its leaves. This study aimed to determine which protocol to use and which part of Ginkgo tree is most appropriate to extract good-quality genomic DNA. For this purpose, cetyltrimethylammonium bromide protocol and protocol of commercially available kit by EZ1 Nucleic acid isolation system have been optimized for extraction of genomic DNA from G. biloba leaves. Efficient yields of high-quality amplifiable DNA was produced rapidly with kit by EZ1 Nucleic acid isolation method. The purified DNA which has excellent spectral quality was efficiently amplified by 5 arbitrary primers (OPA11-15), and was suitable for long-fragment PCR amplification.Öğe Genomic DNA isolation from aromatic and medicinal plants growing in Turkey(ACADEMIC JOURNALS, 2009) Vural, Hasibe CingilliHerbal and aromatic plants are attracting more attention among contemporary plant researchers because some human diseases resulting from bacterial antibiotic resistances have gained worldwide concern. A number of methods are available and are being developed for the isolation of nucleic acids from plants. Because plants contain high amounts of many different substances, it is unlikely that just one nucleic acid isolation method suitable for all plants can ever exist. Therefore, we developed 4 modified new methods that produced good quality DNA from these plants. This article deals with modern approaches in determining genetic variability, in which three categories of genetic markers are applied by morphological, biochemical and molecular. Furthermore, the aim is to assess the available genetic diversity for each species; to provide more accurate and detailed information than is available using classical phenotypic data in this subject. Various types of plant materials and a number of different protocols for the isolation of DNA were tested in order to obtain good quality DNA for PCR reactions. Ten populations of different aromatic and medicinal plants from Turkey were tested in the study. The number of plants examined for each population varied from two to five. When fresh or frozen leaves of plants collected in autumn were used for the isolation of DNA, no positive result in PCR reaction was obtained regardless of the isolation protocol being used. Four different DNA methods were compared for the isolation of DNA from the different plant homogenates, namely the CTAB, Plant Genomic DNA Purification Kit, and EZ1 Nucleic acid isolation methods and DNA extraction with phenol purification and liquid nitrogen method.Öğe Identification of Genotype and Allelic Frequencies of Vitamin D Receptor Gene (Taq1) Polymorphisms in Type 1 Diabetes Mellitus Patients from Turkey(Research Journal Biotechnology, 2012) Vural, Hasibe Cingilli; Isler, OrcunChanges in the DNA or polymorphisms of the VDR cause the protein to bind more or less tightly to 1,25OH. The tighter the vitamin D binds, the stronger and longer lasting the metabolic changes are. Some of the different polymorphisms of the VDR have been associated with an increased risk for Diabetes Mellitus. Recent studies suggest that allelic variations of the vitamin D receptor (VDR) gene can influence Diabetes Mellitus. In brief, the aim of this study was to assess the contribution of these VDR polymorphisms to the susceptibility to Type 1 Diabetes Mellitus in the Turk population. There are several polymorphisms for the VDR gene, but only the three most commonly studied polymorphisms are Taq1, Bsm1 and Fok1 examined. This study suggests that while Taq1 polymorphisms may be functionally different, it may also play a role in serum levels. TT allele of VDR gene has been associated with higher Diabetes Mellitus risk for study on young adults or 100 patients with Type 1 Diabetes Mellitus (DM) (50 women, 50 men) and 120 healthy subjects. The Polymerase chain reaction (PCR) was used for amplification of a 200 bp fragment of the Vitamin D Receptor gene. One study found that TT genotype are overrepresented in Type 1 Diabetes Mellitus patients and those with the TT allele had a 3 fold increase in Type 1 Diabetes Mellitus risk. In addition, the aim of the present study was to adapt PCR amplification, the PCR-RFLP and the most effective DNA isolation method. Taq1 polymorphism indicates susceptibility to Type 1 Diabetes Mellitus in the Turk population. The results indicate that the Taq 1 polymorphism in the VDR gene plays a significant role in protection against T1DM.Öğe Interaction of L-myc oncogene in breast cancer with irinotecan onto functionalized magnetic nanoparticles(ELSEVIER SCIENCE BV, 2013) Maltas, Esra; Ozmen, Mustafa; Vural, Hasibe CingilliThe effect of different functional groups on binding capacity of DNA was studied with bare and modified superparamagnetic iron oxide nanoparticles (SPIONs). For this purpose, modifications were performed with [3(2,3-epoxypropoxy)propyl] trimethoxy silane (GPTS) and (3-aminopropyl) triethoxysilane (APTS) by silanization reaction and also subsequent reaction with glutaraldehyde (GA) to obtain functional groups on nanoparticles surface. L-myc gene from breast tissue with cancer was amplified by using polymerase chain reaction (PCR). APTS functionalized nanoparticles showed the highest binding capacity (82.70%) when compared with bare, GPTS, APTS and GA functionalized SPIONs. DNA immobilized magnetic nanoparticles were also examined by Scanning Electron Microscopy. Binding capacity of irinotecan to DNA onto SPIONs was found as 2.81 x 10(-5) mg/mL for 20 mg of APTS modified nanoparticles by using fluorescence spectroscopy. (C) 2013 Elsevier B.V. All rights reserved.Öğe Investigation of "contagious type antibiotic resistance properties" related with r plasmids in escherichia coli strains isolated from İzmit gulfs (Turkey)(KAFKAS UNIV, VETERINER FAKULTESI DERGISI, 2011) Vural, Hasibe Cingilli; Akcin, AbdulkadirIn this study, 8 Escherichia coli strains one of the pollutions of the Gulf were from the twenty-three seawater samples taken from 9 chosen sites in the Gulf of Izmit, and 8 E. coli coliforms which is one of the pollutions of the Gulf, have been studied with respect to their resistance to antibiotics and their carrier properties of Contagious Types Plasmids with "R factor. E. coli samples were isolated and identified by use of classic methods. Sensitivity tests for antibiotics were conducted according to the Kirby-Bauer Disc Diffussion method. The results of E. coli isolates to various antibiotics were as follows; 50% resistance to Tetracyline, 62.5% for Sulbactam/Amphicillin, 62.5% for Penicillin, 50% for Gentamicin, % 12.5 for Amikacin, 37.5% for Chloramphenicol, 25% for Cefoperozone, 37.5% for Kanamycin, and 62.5% for Trimethoprim + Sulphamethoxozale. This study demonstrates the importance of water pollution in the Gulf of Izmit by these pollution indicators, the presence of coliforms. Furthermore, E. coli a water related bacterium, has been proven to be highly resistant to at least 2 antibiotics and to be able to transfer this resistance to R plasmids. In coliform bacteria, presence of high resistance provider R plasmids have been found. This study may effectively help to understandthe resistance models of the E. coli found at the Gulf of Izmit/Turkey.Öğe İzmit körfezinden izole edilen escherichia coli'lerde r plazmidlerine bağlı "bulaşıcı tipte antibiyotik direnç özelliğinin'' belirlenmesi(2011) Vural, Hasibe Cingilli; Akçin, AbdülkadirBu çalışmada İzmit Körfezi'ndeki 9 istasyondan alınan 23 deniz suyu örneğinden izole edilen ve körfezde kirliliğe neden olan koliformlardan 8 Escherichia coli suşu antibiyotiklere karşı çoklu dirençlilik ve bulaşıcı tipte plazmid "R faktörü'' taşıma özellikleri yönünden incelenmiştir. E. coli suşlarının örneklerden (deniz suyundan) izolasyonu ve identifikasyonu klasik metodlarla yapılmıştır. Antibiyotik duyarlılık testleri Kirby-Bauer Disk Difüzyon Metoduna göre gerçekleştirilmiştir. İzole edilen E. coli suşlarının antibiyotiklere dirençlilikleri Tetracylin için %50, Sulbactam/Ampicillin için %62.5, Penicillin için %62.5, Gentamicin için %50, Amikacin için %12.5, Chloramphenicol için %37.5, Cefoperozone için %25, Kanamycine için %37.5, Trimethoprim Sulphametoxozale için %62.5 olarak tesbit edilmiştir. Bu çalışma, İzmit Körfezi'ndeki kirlenmenin ve kirlilik indikatörü olan koliform varlığının önemli boyutlarda olduğunu göstermektedir. Ayrıca su kaynaklı olan E. coli suşlarının en az 2 antibiyotiğe birden çoklu direnç taşıdıkları ve bu dirençlilik durumlarını R plazmidleri ile aktarabilme yeteneğinde oldukları ispatlanmıştır. Koliform bakterilerde çoklu dirençliliği sağlayan R plazmidlerinin varlığı da saptanmıştır. Bu çalışmanın sonuçları, önemli bir sağlık tehdidi olan İzmit Körfezi'ndeki E. coli suşlarının direnç modelinin ortaya konulmasında önemlidir.Öğe Optimization of DNA isolation for RAPD-PCR analysis of selected (Echinaceae Purpurea L. Moench) medicinal plants of conservation concern from Turkey(ACADEMIC JOURNALS, 2009) Vural, Hasibe Cingilli; Dageri, AsliGenetic analysis of plants relies on high yields of pure DNA samples Here we present the optimization of DNA isolation and PCR conditions for RAPD analysis of selected medicinal plants of conservation concern from Turkey containing high levels of polysaccharides, polyphenols and secondary metabolites The method involves a modified CTAB extraction employing polyvinyl pyrrolidone (PVP) while grinding successive long term chloroform isoamylalcohol extractions, EZ1 nucleic acid isolation protocols The yield of DNA ranged from 1 2 mu g/mu l per gram of the leaf tissue and the purity (ratio) was between 1 7 1 8 indicating minimal levels of contaminating metabolites EZ1 nucleic acid isolation technique is ideal for isolation of DNA from different plant species and the DNA isolated was used for randomly amplified polymorphic DNA (RAPD) analysis RAPD protocol was optimized based on the use of higher concentration of MgCl(2) (3 mM) lower concentrations of primer (0 5 mu M) and Taq polymerase (0 2 units) 50 ng of template DNA and an annealing temperature of 37 degrees C, resulted optimal amplification Reproducible amplifiable products were observed in all PCR reactions Thus the results indicate that the optimized protocol for DNA isolation and PCR was amenable to plant species belonging to different genera which is suitable for further work on diversity analysis Furthermore here we used suitable DNA isolation protocol for RAPD analysis to study the genetic variation in the future in Echinaceae sp grown in TurkeyÖğe Quantification and presence of human ancient DNA in burial place remains of Turkey using real time polymerase chain reaction(ACADEMIC JOURNALS, 2009) Vural, Hasibe CingilliArchaeometry and forensic laboratories are increasingly confronted with problematic samples from the scene of samples, containing only minute amounts of deoxyribonucleic acid ( DNA), which may include polymerase chain reaction (PCR) inhibiting substances. Efficient DNA extraction procedures, as well as accurate DNA quantification methods, are critical steps involved in the process of successful DNA analysis of such samples. Genomic DNA was extracted automatically by using EZ1 Automatic Nucleic Acid Isolation System (Qiagen, Germany) with investigator kit ( Qiagen, Ilden, Germany) from ancient bones. This method is a sensitive for the extraction of DNA from a wide variety of forensic samples, although it is known to be laborious compared with single tube extraction methods. The relatively high DNA recovery and the quality of the extracted DNA speak for itself. For reliable and sensitive DNA quantitation, the application of real time PCR is described. A published real-time PCR assay, which allows for the combined analysis of nuclear or ancient DNA and mitochondrial DNA, was modified. This approach can be used for recovering DNA from the surface of fossil bone remains in Turkey via a simple procedure that permits a direct quantitative and qualitative assessment of molecular markers. Using quantitative RT-PCR, the available sources of total aDNA was shown to consists of intact DNA that is virtually free of RNA, resulting in a more accurate representation of gene expression using RTPCR and PCR amplification methods. In this study, the results demonstrate that RT-PCR method can be useful for the improved ancient DNA extraction in anthropology and archeology.Öğe Radioactivity concentrations and dose assessments of therapeutic peloids from some Turkish spas(MINERALOGICAL SOC, 2015) Karakaya, Muazzez Celik; Dogru, Mahmut; Karakaya, Necati; Vural, Hasibe Cingilli; Kuluozturk, Fatih; Bal, Sultan SahinThe activity concentrations of natural radionuclides in peloids were studied to assess the radiologic hazard from 18 Turkish spas. The peloids are mainly used for therapeutic treatments, rheumatic diseases and aesthetic purposes. The concentrations of the natural radionuclides Ra-226, Th-232, K-40 and Cs-137 were determined with a gamma ray spectrometer using a HPGe detector. The average activity concentrations of Ra-226, Th-232, K-40, and Cs-137 in the peloids studied were 110.69, 71.52, 576.48 and 0.447 Bq/kg, respectively. The radium equivalent activities in the peloid samples ranged from 63.3 to 766.77 Bq/kg. The absorbed dose rate (D-out) varied between 37.52 and 330.67 nGy/h and most of the observed spa doses are greater than the worldwide recommended values. The annual effective dose values range from 0.26 to 2.78 mu Sv/y. The annual gonadal dose equivalents of the samples vary from 224.07 to 2283.55 with a mean of 821.99 mu Sv/y.Öğe Screening of free radical formation in crystals of guanosine by ESR study(ELSEVIER SCIENCE BV, 2011) Usta, Ayhan; Vural, Hasibe Cingilli; Asik, Biray; Usta, KezibanIn this study, to obtain guanosine polycrystalline, novel crystallization method was performed on powder guanosine material. Effective crystallization conditions were achieved by adjustment of the concentration of the metal ions, chemical solutions, NaCl, KCl, glacial acetic acid, nitric oxide, perchloric acid, glutamic acid, and pH of buffer. Behaviors of the guanosine polycrystal samples exposed to high-energy values were investigated using ESR method. The polycrystal samples were exposed to gamma-rays for 48 and 72 h. ESR signals were not recorded from the non-irradiated sample and the sample irradiated for 48 h, but the polycrystalline sample irradiated for 72 h exhibited complex ESR spectra. ESR measurements were taken on the irradiated sample in temperature range from 300 to 450 K. On the basis of all these measurements dependence temperature, it can be said that the shape of the spectrum was to be dependent on temperature slightly. Hence, we assume that the radical structure occurred was resistance to high temperature. Two radicals were determined in the structure irradiated and these were called radical I and radical II. The g, hyperfine constants, and spin density were found to be rho = 0.96, a(NNH)(N) = 2.7 mT, a(NNH)(N) = 1.155 mT, a(N) = 0.35 ml and g(1) = 2.0093 for the radical 1; a(N) = 4.7 ml and g(2) = 2.0094 for the radical II. (C) 2011 Elsevier B.V. All rights reserved.Öğe Species Determination of Ancient Bone DNA From Fossil Skeletal Remains of Turkey Using Molecular Techniques(ACADEMIC JOURNALS, 2010) Vural, Hasibe Cingilli; Tırpan, Ahmet AdilThe use of genetic technology in forensic science and archaeometry is applied primarily to distinguish between individials who may be the source of biological material associated with archeological remains. DNA sequences from ancient fossils have great potential for studies of phylogeny, biogeography and molecular evolution. DNA from fossils also facilitates the rigorous testing and calibration of mutation rates among related taxa, sex test and molecular divergence time (Cano et al., 1993; Burger et al., 1999). In this study, a rapid and quantitative ancient DNA extaction methods from human skeletal remains was developed for application of forensic science and archaeometry. For that reason, DNA was extracted from ancient human bones from Mugla in Turkey. Furthermore, all the bone samples which are obtained from burial place are subjected to DNA isolation and then interspecific sequence polymorphisms in the mitochondrial cytochrome b gene were analyzed by PCR to determine the species origin of Bronze Age animal and human skeletal remains. Existing techniques were refined by targeted primer design focusing on a DNA fragment shorter than 200 bp, an approach allowing us to identify up to all bone samples at the same time. For routine applications in archaeometry, food or material analyses, PCR may thus provide a simple alternative to sequencing of PCR products, allowing discrimination between species, even if the template DNA is degraded or contains traces of DNA from various species.
