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    Antiprotozoal Activity of Artemisia vulgaris and Berberis vulgaris Against Leishmania major and Trichomonas vaginalis
    (Springer Int Publ Ag, 2026) Aksoy, Tulay; Girginkardesler, Nogay; Balcioglu, Ibrahim Cuneyt; Kilimcioglu, Ali Ahmet
    Purpose Leishmania major and Trichomonas vaginalis infections pose a significant global health burden, while current treatments are limited by toxicity, resistance, and restricted accessibility. This study aimed to evaluate the in vitro and ex vivo antileishmanial effects of Artemisia vulgaris and Berberis vulgaris extracts against L. major, as well as their in vitro antitrichomonal activity against T. vaginalis trophozoites. Methods Ethanolic extracts of A. vulgaris and B. vulgaris were tested against L. major promastigotes, intracellular amastigotes, and T. vaginalis trophozoites. Parasite viability was determined by CellTiter-Glo (R), microscopy, and rescue-transformation assays, and selectivity indices (SI) were calculated. Amphotericin B and metronidazole served as reference drugs. Results Both extracts exhibited low cytotoxicity in THP-1 macrophages (A. vulgaris CC50 = 465.2 & micro;g/mL; B. vulgaris = 357.7 & micro;g/mL). Against L. major, B. vulgaris showed greater potency (IC50 = 76.8 & micro;g/mL; SI = 4.7 for amastigotes) than A. vulgaris (IC50 = 179.7 & micro;g/mL; SI = 2.6). Both extracts reduced intracellular parasite burden in a dose-dependent manner, achieving complete clearance at non-cytotoxic concentrations (>= 300 & micro;g/mL). In T. vaginalis, the extracts induced concentration-dependent inhibition, with IC50 values of 68.9 & micro;g/mL (B. vulgaris, SI = 5.2) and 104.4 & micro;g/mL (A. vulgaris, SI = 4.5). Conclusion Both extracts exhibited selective, dose-dependent antiprotozoal activity, with B. vulgaris showing superior efficacy, particularly against intracellular L. major and T. vaginalis. These results highlight their potential as natural antiprotozoal sources, warranting further studies on active constituents, mechanisms, and in vivo efficacy.
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    Investigation of Anti-leishmanial Effects of Bee Products (Honey, Propolis) on Leishmania tropica Promastigotes
    (Ankara Microbiology Soc, 2020) Aksoy, Tulay; Sivcan, Eda; Dogan, Fatma; Cetin, Songul; Yar, Turkan Mutlu
    This study was aimed to investigate the anti-leishmanial effects of bee products (honey and propolis) by using the causative agent of cutaneous leishmaniasis Leishmania tropica promastigotes, in in vitro culture. In vitro anti-leishmanial efficacy of honey (pine, flower and chestnut) and propolis used in the study were evaluated using the microdilution method. Honey, which is a bee product, was dissolved with RPMI medium containing fetal calf serum (FCS) and diluted in the same medium, and serial dilutions were prepared in concentrations between 62.5-1000 mg/ml. Propolis, on the other hand, was dissolved with ethyl alcohol and only 2.5 mu l was used from all these concentrations since the alcohol content was more than 50% in these concentrations prepared and we thought that this rate would negatively effect the parasite development. Then, RPMI containing FCS was diluted in the medium and serial dilutions were prepared at concentrations between 50-800 mu g/ml. To the dilutions prepared, the promastigot suspension was added so that their final concentrations in the wells were 1 x 10(6) promastigot/ml and then the medium was incubated for 24 and 48 hours in 26 degrees C. After the incubation, promastigotes were determined microscopically for morphology, mobility and live parasite density, and cell viability was determined by MTS method and 50% inhibitor concentrations (IC50) were compared with control groups. Anti-leishmanial activity of propolis (50, 100, 200, 400 and 800 mu g/ml) and honey (62.5, 125, 250, 500 and 1000 mg/ml) on promastigotes was evaluated in vitro. In microscopic examinations, pine honey showed anti-leishmanial activity starting from 62.5 mg/ml, flower honey 250 mg/ml, and chestnut honey 125 mg/ml, and pine honey was more effective on promastigotes (p< 0.05), and propolis was effective from 100 mu g/ml concentration. It has been determined that very low concentrations of propolis caused changes in the morphological structure of the parasites and were more effective than the other bee products. The prevention of cell proliferation and decreasing of the IC50 values according with the time of pine honey (IC50= 109.28 mg/ml), flower honey (IC50= 248.07 mg/ml), chestnut honey (IC50= 147.65 mg/ml) and propolis (IC50= 82.98 pg/ml) applied on L.tropica promastigot cell culture was determined by MTS method. In this study, it was found that various concentrations of pine, flower, chestnut honey and propolis showed anti-leishmanial activity on L. tropica promastigotes. It has been observed that pine honey is more effective on promastigotes after 48 hours of incubation period, and propolis is more effective in both morphology and cell inhibition of the parasites even at very low concentrations. It is believed that these data can be used as an alternative treatment method against cutaneous leishmaniasis infections and further studies are required.
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    Molecular Characterization of Lucimycin Gene of Lucilia sericata
    (Ankara Microbiology Soc, 2020) Cetin, Songul; Aksoy, Tulay
    Lucilia sericata, a member of the Calliphoridae family, is one of the most common species in the genus Lucilia. Medical importance of L.serkata stems from its use in maggot debridement therapy (MDT). MDT is the name of L.sericata larvae being sterilized and used in the treatment of non-healing wounds. L.sericata maggots used in the treatment of chronic and non-healing wounds (decubitus ulcer, venous leg ulcer, diabetic foot ulcer, etc.) clean the wounds with the help of secreted proteolytic trypsin and lucimycin-like enzymes. The aim of the study was to determine the molecular characterization of lucimycin gene obtained from L.sericata larvae in MDT by using molecular methods and to contribute to the literature. In this study, continuous production of adult colonies of L.sericata species was carried out in insectarium unit where conditions such as light, humidity and temperature were formed. The life cycle of L.sericata was followed and the production of eggs, larvae, pupae, adult flies and fly colonies of the species were formed. In the third stage larvae obtained from adult flies in the insectarium unit, RNA was isolated and subsequently cDNA synthesis was performed by reverse transcription. Polymerase chain reaction (PCR) analysis of the synthesized cDNAs with the specific primers designed for the lucimycin gene of L.sericata was performed and the obtained amplicons were cloned into pJET1.2/blunt vector and the plasmid was purified. The recombinant plasmids were sequenced with vector-specific primers and target gene region sequences were obtained. After the molecular characterization of the isolate with nucleotide sequences was determined, it was registered to GenBank database with the accession number MF964229. The PCR product of 288 bp was obtained from the cDNA obtained from the larvae of L.sericata produced in the insectarium unit by PCR using lucimycin specific primers. The PCR product imaged on the gel was purified by transformation and subsequent colonies were screened to see whether they contained recombinant plasmids. Three of the colonies were identified as recombinant plasmids containing L.sericata lucimycin gene by PCR screening. From three colonies confirmed by PCR screening, recombinant plasmids containing L.sericata lucimycin gene were purified by miniprep. The recombinant plasmid product was confirmed to contain the L.sericata lucimycin gene by PCR from a total of 20 mu l of the recombinant plasmid miniprep product. DNA sequencing analysis was performed to confirm the plasmid after cloning. The 288 bp L.sericata lucimycin sequence was confirmed by DNA sequence analysis. The lucimycin gene isolated was confirmed by specific and pJET1.2 forward and reverse primers using Blastn algorithm as a result of species and/or subspecies using the Blastn algorithm and the related isolate was recorded in GenBank database with the MF964229 accessory number. The DNA sequence of the isolated sample was compared with other isolates found in GenBank by Pubmed/Blast program. KJ413251.1 was found to be 99% similar to the GenBank isolate. The 113th nucleotide was C (cytosine) in the sequence of our isolate, while the existence of G (guanine) in the sequence numbered KJ413251.1 GenBank revealed the difference between the two sequences. In this study the molecular characterization of lucimycin gene derived from L. sericata larvae were determined for the first time in Turkey, it is assumed that this molecule which has an antifungal property, can be used in the studies that will be carried out in the future, especially in microorganisms causing cutaneous infections. The study is important since the isolate is registered as a biological asset of Turkey in GenBank and also being the second study in the world.
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    Thymol's antileishmanial activity and its impact on host cytokine profiles: In vitro and ex vivo studies on Leishmania tropica
    (Elsevier Ireland Ltd, 2026) Aksoy, Tulay; Kilimcioglu, Ali Ahmet
    Cutaneous leishmaniasis (CL) is a neglected tropical disease associated with significant morbidity, primarily due to chronic skin lesions, scarring, and psychosocial consequences. This study aimed to investigate the in vitro and ex vivo antileishmanial effects of thymol (1-500 mu M) against Leishmania tropica (MHOM/TR/2012/CBCL-LT) infection. Thymol's in vitro efficacy was assessed on both promastigote (Haemocytometry and CellTiter-Glo assays) and amastigote (Giemsa staining and Parasite Rescue Transformation Assay) forms of L. tropica. Additionally, its immunomodulatory effects were evaluated by analyzing cytokine secretion (IFN-gamma, IL-12, IL-10, and IL-4) and infectivity in THP-1 macrophages using ELISA. Cytotoxicity was determined by calculating the 50 % cytotoxic concentration (CC50) in THP-1 cells. The in vitro inhibitory concentration (IC50) value against L. tropica promastigotes was determined as 79.41 mu M, while the ex vivo IC50 value against amastigotes was 105.2 mu M. Incubation of infected macrophages with thymol resulted in a dose-dependent increase in IFN-gamma and IL-12 levels, along with a significant reduction in IL-10 and IL-4 secretion (p < 0.05). The CC50 value of thymol in THP-1 cells was 160.7 mu M, indicating low cytotoxicity. Moreover, the selectivity index (SI) values greater than 1 confirmed the compound's preferential action against amastigotes while exhibiting minimal toxicity toward macrophages. These findings highlight thymol's potential as an antileishmanial agent by effectively eliminating and controlling Leishmania parasites in both in vitro and ex vivo models. Due to its immunomodulatory properties and low cytotoxicity, thymol represents a promising starting point for the development of novel antileishmanial agents and alternative therapeutic strategies against CL caused by L. tropica.