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Öğe Antifungal and Antibiofilm Activities of Selective Serotonin Reuptake Inhibitors Alone and in Combination with Fluconazole(Turkish Pharmacists Assoc, 2020) Tekintas, Yamac; Temel, Aybala; Ates, Aysegul; Erac, Bayri; Metin, Dilek YeIm; Polat, Suleyha Hilmioglu; Limoncu, Mine HosgorObjectives: Candida spp. are clinically important pathogens that cause difficulties for treatment by biofilm formation. Considering antifungal resistance rates and the limitations in the discovery of new antifungals, the antifungal and antibiofilm effects of various drugs used for different therapeutic purposes are becoming more important. The goal of our study was to determine the antifungal and antibiofilm effects of the selective serotonin reuptake inhibitors (SSRIs), namely sertraline (SRT), paroxetine (PRX), and fluoxetine (FLX) alone and in combination with fluconazole (FLC) against Candida spp. Materials and Methods: Twenty Candida spp. strains isolated from clinical samples from Ege University Hospital were identified by the Dalmau method and matrix-assisted laser desorption ionization time of flight mass spectrometry. The minimum inhibitory concentrations (MICs) of the SSRIs and FLC were detected by broth microdilution method. Synergistic interactions between the SSRIs and FLC were investigated by checkerboard assay. The antibiofilm effects of the SSRIs were determined by spectrophotometric microplate method. Results: Among the isolates, five different Candida spp. (C. albicans, C. glabrata, C. krusei, C. tropicalis, and C.parapsilosis) were identified. The MICs of the SSRIs ranged between 16-512 mu g/mL. While SRT showed the highest antifungal effect, the antibiofilm efficacy of FLX was higher than that of the other agents. Moreover, FLX and PRX showed a synergistic effect with FLC in 13 and 19 isolates, respectively. Four isolates were strong biofilm producers while nine isolates were moderate biofilm producers. C. parapsilosis strains showed higher biofilm production than the other species. At MIC/2 concentration, FLX and SRT alone inhibited mature biofilms in six and five isolates, respectively, while PRX caused increases biofilm formation in seven isolates. Conclusion: This study revealed that MIC/2 concentrations of SSRIs could have antifungal and antibiofilm effects. SRT and FLX alone or in combination with antifungals may possibly have therapeutic potential for combating fungal infections.Öğe Antifungal and Antibiofilm Activities of Selective Serotonin Reuptake Inhibitors Alone and in Combination with Fluconazole(2020) Temel, Aybala; Tekintaş, Yamaç; Polat, Süleyha Hilmioğlu; Metin, Dilek Yeşim; Ateş, Ayşegül; Eraç, Bayrı; Limoncu, Mine HoşgörObjectives: Candida spp. are clinically important pathogens that cause difficulties for treatment by biofilm formation. Considering antifungalresistance rates and the limitations in the discovery of new antifungals, the antifungal and antibiofilm effects of various drugs used for differenttherapeutic purposes are becoming more important. The goal of our study was to determine the antifungal and antibiofilm effects of the selectiveserotonin reuptake inhibitors (SSRIs), namely sertraline (SRT), paroxetine (PRX), and fluoxetine (FLX) alone and in combination with fluconazole(FLC) against Candida spp.Materials and Methods: Twenty Candida spp. strains isolated from clinical samples from Ege University Hospital were identified by the Dalmaumethod and matrix-assisted laser desorption ionization time of flight mass spectrometry. The minimum inhibitory concentrations (MICs) of the SSRIsand FLC were detected by broth microdilution method. Synergistic interactions between the SSRIs and FLC were investigated by checkerboardassay. The antibiofilm effects of the SSRIs were determined by spectrophotometric microplate method.Results: Among the isolates, five different Candida spp. (C. albicans, C. glabrata, C. krusei, C. tropicalis, and C.parapsilosis) were identified. The MICsof the SSRIs ranged between 16-512 µg/mL. While SRT showed the highest antifungal effect, the antibiofilm efficacy of FLX was higher than that ofthe other agents. Moreover, FLX and PRX showed a synergistic effect with FLC in 13 and 19 isolates, respectively. Four isolates were strong biofilmproducers while nine isolates were moderate biofilm producers. C. parapsilosis strains showed higher biofilm production than the other species.At MIC/2 concentration, FLX and SRT alone inhibited mature biofilms in six and five isolates, respectively, while PRX caused increases biofilmformation in seven isolates.Conclusion: This study revealed that MIC/2 concentrations of SSRIs could have antifungal and antibiofilm effects. SRT and FLX alone or incombination with antifungals may possibly have therapeutic potential for combating fungal infections.Öğe Bacterial Biofilms: Detection Methods and Role in Antibiotic Resistance(2018) Temel, Aybala; Eraç, Bayrı…Öğe Bazı ilaç moleküllerinin klinik acinetobacter baumannii biyofilm yapısı üzerine etkilerinin gerçek zamanlı hücre analizi yöntemi ile incelenmesi(Ege Üniversitesi, Sağlık Bilimleri Enstitüsü, 2019) Temel, Aybala; Eraç, BayrıAcinetobacter baumannii, ventilatör ilişkili pnömoni ve kateter ilişkili bakteriyemi gibi ciddi nozokomiyal enfeksiyonlara neden olabilen, önemli bir fırsatçı patojendir. Bu bakterinin biyofilm oluşturma özelliği gibi virülans faktörleri ve son yıllarda hızla artan antibiyotik direnci, patogeneze katkı sağlayarak, enfeksiyonların tedavisini zorlaştırmaktadır. Farklı ülkelerde yürütülen çok sayıda araştırma, çoklu ilaç dirençli A.baumannii izolatlarının artışına ve bu izolatlarda biyofilm üretim kapasitesinin de yüksek olduğuna işaret etmektedir. Bu açıdan, antibakteriyel ve antibiyofilm etkinliğe sahip ilaç veya ilaç kombinasyonlarına yönelik araştırmalar değer kazanmaktadır. Ayrıca, tedavi başarısızlıklarında önemli bir rolü olan biyofilm oluşumunun hızlı ve doğru bir şekilde belirlenmesi, enfeksiyonlarla mücadele açısından kritik bir öneme sahiptir. Çalışmamızda, tigesiklin (TGC), n-asetilsistein (NAS) ve asetilsalisilik asit (ASA) gibi çeşitli ilaç moleküllerinin, klinik A. baumannii izolatlarının biyofilm yapısı üzerine etkilerinin, konvansiyonel ve gerçek zamanlı hücre analizi (Real Time Cell Analyses-RTCA) yöntemleri ile incelenmesi amaçlanmıştır. Ayrıca, incelenen etken maddelerin A. baumannii suşlarında biyofilm ile ilişkili bazı genlerin ekspresyon seviyelerine etkisi de araştırılmıştır. Ege Üniversitesi Hastanesi’nden izole edilen ve klonal olarak ilişkisiz 22 A.baumanni izolatının biyofilm oluşturma kapasiteleri öncelikle spektrofotometrik mikroplak-kristal viyole yöntemi ile belirlendi. Güçlü biyofilm üreticisi olduğu belirlenen altı temsilci kökenin biyofilm oluşturma kapasiteleri RTCA yöntemi ile de araştırıldı ve elde edilen sonuçların konvansiyonel yöntem ile uyumlu olduğu saptandı. RTCA yöntemi ile 8-10 saat içinde güçlü biyofilm üreticisi kökenler ile zayıf üretici ve negatif kontroller arasında istatistiksel olarak anlamlı fark bulundu. Tigesiklin, NAS ve ASA moleküllerinin tek başlarına ve kombine halde, kökenlerin biyofilm oluşumu ve olgun biyofilmleri üzerine etkileri her iki yöntemle de değerlendirildi ve sub-MİK konsantrasyonlarda TGC+NAS ve TGC+ASA kombinasyonlarının sinerjik etkili olduğu tespit edildi. İncelenen etken maddelerin A. baumannii kökenlerinin biyofilm oluşumu üzerine etkilerini saptamak açısından konvansiyonel ve RTCA yöntemleri paralel sonuçlar verirken, olgun biyofilm üzerine etkilerin değerlendirilmesinde RTCA yöntemi ile herhangi bir etki belirlenemedi. Temsilci A.baumannii kökenlerinde biyofilm oluşumu ile ilişkilendirilen bazı virülans genlerinin (bap, adeG, csuE, ompA) varlığı polimeraz zincir reaksiyonu ile araştırıldı. Altı kökenin tamamında csuE ve ompA genleri tespit edilirken, ikişer kökende bap ve adeG gen varlığı gözlendi. Fenotipik yöntemlerle sinerjik etkili olduğu saptanan ilaç kombinasyonlarının, bazı suşlarda csuE ve bap gen ekpresyon seviyelerinde düşüşe neden olduğu kantitatif gerçek zamanlı ters transkriptaz kantitatif polimeraz reaksiyonu (RT-qPCR) ile saptandı. Bildiğimiz kadarıyla, A. baumannii kökenlerinde çeşitli ilaç moleküllerinin biyofilm yapısı üzerine etkilerinin RTCA yöntemi ile ilk olarak değerlendirildiği çalışmamızda, genel olarak konvansiyonel yöntem ile uyumlu sonuçlar alınmış, sub-MİK konsantrasyonlarda sinerjik etkili kombinasyonlar saptanmıştır. Ayrıca, incelenen etken maddelerin A. baumannii biyofilmi üzerinde görülen fenotipik etkilerinin, biyofilm ilişkili bazı genlerinin ekspresyon seviyelerindeki azalmaya bağlı olabileceği de ortaya konmuştur. RTCA yöntemi biyofilm üreticisi kökenleri saptamada hızı ve tekrar edilebilirliği ile öne çıkarken, etken maddelerin olgun biyofilm üzerine etkilerinin değerlendirilmesi için uygun gözükmemektedir.Öğe COVID-19 Pandemisinde Mikrobiyolojik Tanı Yöntemleri(2021) Temel, Aybala; Ateş, Ayşegül; Eraç, BayrıDünya genelinde bir milyondan fazla insanın ölümüne neden olan COVID-19 pandemisi; enfeksiyon hastalıklarının önlenmesi ve salgınların kontrol altına alınmasında mikrobiyolojik tanı yöntemlerinin kritik önemini açıkça göstermiştir. SARS-CoV-2 ile mücadelede en etkili yol, virüsün yayılımını olabildiğince azaltmaktır. Virüsün yayılımını azaltmak ise virüsü taşıyan semptomatik veya asemptomatik tüm bireylerin, en kısa sürede ve doğru şekilde saptanmasını sağlayan tanı yöntemleriyle olasıdır. COVID-19 hastalığının tanısında ve takibinde, uluslararası sağlık otoriteleri tarafından önerilen çeşitli mikrobiyolojik yöntemler (serolojik testler, nükleik asit amplifikasyon yöntemleri) yaygın olarak kullanılmaktadır. Dünya Sağlık Örgütü tarafından COVID-19 laboratuvar tanısında altın standart yöntem olarak belirtilen gerçek zamanlı polimeraz zincir reaksiyonu (RT-PZR) metodu dâhil olmak üzere farklı diagnostik yöntemlerin uygulanmasında yaşanan zorluklar pandemi sürecinde bir kez daha anlaşılmıştır. Enfeksiyonlarla mücadelede daha yüksek başarı sağlayabilmek için mevcut mikrobiyolojik tanı yöntemlerinin uygulama ve değerlendirme standartlarının iyi belirlenmesi, daha kısa sürede doğru şekilde sonuç veren, kolay uygulanabilen, düşük maliyetli yeni yöntemlerin geliştirilmesi gerekmektedir.Öğe In vitro effects of antibiofilm agents and antibiotics on coagulase-negative staphylococci(Marmara Univ, 2020) Ozturk, Ismail; Tekintas, Yamac; Temel, Aybala; Ermertcan, Safak; Kurutepe, Semra; Hosgor Limoncu, MineCoagulase negative staphylococci (CoNS) are important nosocomial pathogens that cause biofilm infections. Biofilm provides advantages for microorganisms to resist antibiotics and host immune systems. Considering the increased antibiotic resistance, alternative treatments are needed to combat biofilm infections. in the present study, the effects of antibiotics including gentamicin (GEN), ciprofloxacin, doxycycline (DOX), rifampicin (RIF) and antibiofilm agents including N-acetylcysteine (NAC), ethylenediaminetetraaceticacid (EDTA), nisin (NIS), farnesol (FAR) on clinical CoNSbiofilm and IS256, icaA gene expression levels were evaluated. Forty-five CoNS strains were isolated from patients' catheters, at Manisa Celal Bayar University Hospital. The minimum inhibitory concentrations (MICs) of agents were detected by broth microdilution method with European Committee for Antimicrobial Susceptibility Testing (EUCAST) criteria. The combined effects of agents were investigated by checkerboard method. The antibiofilm effects of combinations were investigated by spectrophotometric microplate method. The effects of combinations on IS256 and icaA gene expressions were evaluated by real-time quantitative reverse-transcriptase PCR. Twenty-four isolates (53.3%) were detected as strong biofilm producer. Biofilm production was inhibited in seven isolates in the presence of EDTA+RIF and NIS+DOX while NIS+GEN combination and RIF inhibited biofilm in six isolates. Nine combinations were found to have synergistic effect against isolate #6 which are resistant to four different antibiotics. The expressions of icaA and IS256 were downregulated in the presence of EDTA, NAC+CIP, NAC+GEN, NIS+GEN, FAR+GEN. Antibiofilm agent/antimicrobial combinations could have promising effects for preventing catheter colonization. The further studies on antibiofilm treatment strategies would be beneficial for decreasing morbidity-mortality rates and healthcare costs caused by biofilms.Öğe In vitro effects of antibiofilm agents and antibiotics oncoagulase-negative staphylococci(2020) Ermertcan, Şafak; Limoncu, Mine Hoşgör; Tekintaş, Yamaç; Kurutepe, Semra; Öztürk, İsmail; Temel, AybalaCoagulase negative staphylococci (CoNS) are important nosocomial pathogens that cause biofilm infections. Biofilm provides advantages for microorganisms to resist antibiotics and host immune systems. Considering the increased antibiotic resistance, alternative treatments are needed to combat biofilm infections. In the present study, the effects of antibiotics including gentamicin (GEN), ciprofloxacin, doxycycline (DOX), rifampicin (RIF) and antibiofilm agents including N-acetylcysteine (NAC), ethylenediaminetetraaceticacid (EDTA), nisin (NIS), farnesol (FAR) on clinical CoNS biofilm and IS256, icaA gene expression levels were evaluated. Forty-five CoNS strains were isolated from patients’ catheters, at Manisa Celal Bayar University Hospital. The minimum inhibitory concentrations (MICs) of agents were detected by broth microdilution method with European Committee for Antimicrobial Susceptibility Testing (EUCAST) criteria. The combined effects of agents were investigated by checkerboard method. The antibiofilm effects of combinations were investigated by spectrophotometric microplate method. The effects of combinations on IS256 and icaA gene expressions were evaluated by real-time quantitative reverse-transcriptase PCR. Twenty-four isolates (53.3%) were detected as strong biofilm producer. Biofilm production was inhibited in seven isolates in the presence of EDTA+RIF and NIS+DOX while NIS+GEN combination and RIF inhibited biofilm in six isolates. Nine combinations were found to have synergistic effect against isolate #6 which are resistant to four different antibiotics. The expressions of icaA and IS256 were downregulated in the presence of EDTA, NAC+CIP, NAC+GEN, NIS+GEN, FAR+GEN. Antibiofilm agent/antimicrobial combinations could have promising effects for preventing catheter colonization. The further studies on antibiofilm treatment strategies would be beneficial for decreasing morbidity-mortality rates and healthcare costs caused by biofilms.Öğe Investigating Biofilm Formation and Antibiofilm Activity Using Real Time Cell Analysis Method in Carbapenem Resistant Acinetobacter baumannii Strains(Springer, 2022) Temel, Aybala; Erac, BayriAcinetobacter baumannii is a significant nosocomial pathogen, with its biofilm forming capacity playing an important role in its pathogenicity. The fast and reliable detection of the biofilm formation and measurement of antibiofilm activity of various molecules are critical for combating A. baumannii infections. In this study, we aimed to detect biofilm formation by real time cell analyses (RTCA) method in clinical A. baumannii isolates and to investigate antibiofilm activities of tigecycline (TGC), N-acetylcysteine (NAC), and acetylsalicylic acid (ASA). The effect of the tested drugs on expressions of biofilm-related genes bap and csuE in clinical A. baumannii strains was also analyzed by real time quantitative reverse transcription polymerase chain reaction (RT-qPCR). Biofilm forming capacities for strong and weak biofilm producer A. baumannii strains were detected within 10 h by RTCA method (P < 0.05). We also observed that sub-minimum inhibitory concentrations of NAC + TGC and ASA + TGC combinations could significantly reduce biofilm formation and expression of biofilm-related genes in A. baumanii strains. No statistically significant activity of the tested drugs was detected against mature biofilms of the bacterial strains with the RTCA method. These results suggest that reproducible results on biofilm production capacity of A. baumannii strains and antibiofilm activities of various compounds can be obtained in a short time using RTCA method. Therefore, RTCA method seems to be a beneficial technique for biofilm detection and can help in combating A. baumannii infections by giving health providers the opportunity of implementing antibiofilm treatment strategies in a timely manner.Öğe Tc-99m-Labeled, Colistin Encapsulated, Theranostic Liposomes for Pseudomonas aeruginosa Infection(Springer, 2023) Karpuz, Merve; Temel, Aybala; Ozgenc, Emre; Tekintas, Yamac; Erel-Akbaba, Gulsah; Senyigit, Zeynep; Atlihan-Gundogdu, EvrenInfectious diseases are still the major issue not only due to antibiotic resistance but also causing deaths if not diagnosed at early-stages. Different approaches including nanosized drug delivery systems and theranostics are researched to overcome antibiotic resistance, decrease the side effects of antibiotics, improve the treatment response, and early diagnose. Therefore, in the present study, nanosized, radiolabeled with Tc-99m, colistin encapsulated, neutral and cationic liposome formulations were prepared as the theranostic agent for Pseudomonas aeruginosa infections. Liposomes exhibited appropriate physicochemical properties thanks to their nano-particle size (between 173 and 217 nm), neutral zeta potential value (about - 6.5 and 2.8 mV), as well as encapsulation efficiency of about 75%. All liposome formulations were radiolabeled with over 90% efficiency, and the concentration of stannous chloride was found as 1 mg.mL(-1) to obtain maximum radiolabeling efficiency. In alamar blue analysis, neutral liposome formulations were found more biocompatible compared with the cationic formulations. Neutral colistin encapsulated liposomes were found to be more effective against P. aeruginosa strain according to their time-dependent antibacterial effect, in addition to their highest bacterial binding capacity. As conclusion, theranostic, nanosized, colistin encapsulated, neutral liposome formulations were found as promising agents for the imaging and treating of P. aeruginosa infections.
