Browsing by Author "Akyar I."
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Item Feconomics®: A simple, novel and fast technique for stool concentration in parasitology laboratory; [Feconomics®: Parazitoloji laboratuvarında dışkının konsantrasyonu için basit, yeni ve hızlı bir yöntem](Veteriner Fakultesi Dergisi, 2012) Kurt Ö.; Akyar I.; Görgün S.; Kocagöz T.; Özbilgin A.Feconomics® is a new ready-to-use kit for fecal concentration that eliminates the need for centrifugation and floatation by using absorbent beads. To assess its efficacy in the diagnosis of intestinal parasites, a comparative, double-blind study was conducted in the Parasitology Laboratory of Celal Bayar University Medical School. Stools (Group I, n=251) submitted for routine ova and parasite examination were concentrated with both routine formalin ethyl acetate concentration (FEAC) technique and Feconomics®. Since the number of helminthes identified in the stool samples of patients were very low, helminthes obtained from the animal models in the laboratory were included (Group II, n=11). The iodine-stained samples of all stools and some of the positive samples stained with Gomori’s trichrome and Kinyoun’s acid fast stain were read by specialists. In Group I, 103 of 251 (41.04%) samples were found to be positive for one or more intestinal parasites; among them, 76 (30.28%) and 96 (38.25%) stools were found to be positive with FEAC and Feconomics®, respectively, and the difference was significant (P=0.000). Same parasites were identified with both methods among all 11 samples in Group II. There was no difference between the methods for the morphological integrity and visual appearances of the parasites having cyst or egg forms; yet, it was noticed that the vegetative forms of the parasites were only identified with Feconomics®. Review of our data indicated that Feconomics® may be suggested as a fast and effective fecal concentration method for Parasitology laboratories owing to the identification of higher number of parasites compared to FEAC, and parasites with only vegetative forms such as Dientamoeba fragilis. © 2012, Veteriner Fakultesi Dergisi. All rights reserved.Item Leishmaniasis in Turkey: Determination of Leishmania species by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS)(Tehran University of Medical Sciences (TUMS), 2014) Culha G.; Akyar I.; Yildiz Zeyrek F.; Kurt Ö.; Gündüz C.; Özensoy Töz S.; Östan I.; Cavus I.; Gülkan B.; Kocagöz T.; Özbel Y.; Özbilgin A.Background: Cutaneous leishmaniasis (CL) is endemic in Southeastern Anatolia, mainly in Sanliurfa and Hatay provinces, and the causative agents are mostly Leishmania tropica and less frequently L. infantum. Here, we report the first MALDI-TOF analyses of Leishmania promastigotes obtained from the cultures of two CL cases from Osmaniye and Hatay provinces who were initially diagnosed by microscopy, culture and identified as L. infantum with Real-Time PCR (RT-PCR). Methods: Samples obtained from the skin lesions of patients were initially stained with Giemsa and cultivated in NNN medium. Examination of the smears and cultures revealed Leishmania amastigotes and promastigotes, respectively. The promastigotes (MHOM/TR/2012/CBU15 and MHOM/TR/2012/MK05) obtained from the cultures of both patients were used for RT-PCR targeting the ITS-1 region in the SSU of rRNA. The reference strains of four Leishmania species (L. infantum, L. donovani, L. tropica and L. major) were initially assessed with MALDI-TOF and their data were added to MALDI-TOF Biotyper Library. Results: Both RT-PCR and MALDI-TOF analyses indicated that the causative agent in both patient samples was L. infantum. Conclusion: Despite disadvantages such as requirement of culture fluid with nothing but promastigotes and high cost, MALDI-TOF analysis may be a fast, sensitive and specific diagnostic tool in especially large-scale research studies, where the cost declines, relatively.Item Leishmaniasis in Turkey: First clinical isolation of Leishmania major from 18 autochthonous cases of cutaneous leishmaniasis in four geographical regions(Blackwell Publishing Ltd, 2016) Özbilgin A.; Çulha G.; Uzun S.; Harman M.; Topal S.G.; Okudan F.; Zeyrek F.; Gündüz C.; Östan I.; Karakuş M.; Töz S.; Kurt Ö.; Akyar I.; Erat A.; Güngör D.; Kayabaşi Ç.; Çavuş I.; Bastien P.; Pratlong F.; Kocagöz T.; Özbel Y.Objective: To report isolation of Leishmania major strains obtained from 18 Turkish autochthonous cutaneous leishmaniasis (CL) patients infected with L. major between 2011 and 2014. Methods: Initial diagnosis relied on microscopy and culture in enriched medium, prepared by adding specific amounts of liver extract, protein and lipid sources to NNN medium. Promastigotes were then transferred to RPMI medium including 10% of foetal calf serum for mass culture. Species-specific real-time PCR targeting ITS1 region of Leishmania spp. was performed using both lesion aspiration samples and cultured promastigotes. Two of 18 isolates were identified by isoenzyme analysis in the Leishmaniasis Reference Center in Montpellier, France. Each isolate was inoculated into the footpads of six mice to observe the pathogenicity of L. major. Developing lesions were observed, and the thickening of footpads was measured weekly. Results: Melting curve analyses of 18 isolates showed a peak concordant with L. major, and two of them were confirmed by isoenzyme analyses as L. major zymodeme MON103. In the mouse model, acute lesions seen on day 21 were accepted as an indication of heavy infection. Severe impairments were observed on all mouse footpads over 3 weeks, which even progressed to extremity amputation. Conclusion: Cutaneous leishmaniasis-causing L. major was recently identified in Adana province in southern Turkey, with PCR. Our study shows that such CL cases are not limited to Adana but currently present from western to Southeastern Anatolia, and along the Mediterranean coast. The role of small mammals, the main reservoirs of L. major in Anatolia, needs to be elucidated, as do the underlying factors that cause severe clinical manifestations in L. major infections in Turkey, contrary to the infections in neighbouring countries. © 2016 John Wiley & Sons Ltd.Item Evaluation of the performance of maldi-tof ms and dna sequence analysis in the identification of mycobacteria species(Turkiye Klinikleri Journal of Medical Sciences, 2018) Akyar I.; Çavuşoğlu C.; Ayaş M.; Sürücüoğlu S.; İlki A.; Kaya D.E.; Beşli Y.Background/aim: Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry is an alternative way of identifying mycobacteria via the analysis of biomolecules. It is being increasingly used in routine microbiology practice since it permits early, rapid, and cost-effective identification of pathogens of clinical importance. In this study, we aimed to evaluate the efficacy of phenotypic identification of mycobacteria by the MALDI-TOF MS MBT Mycobacteria Library (ML) 4.0 (Bruker, Daltonics) compared to standard sequence analysis. Materials and methods: A total of 155 Mycobacterium clinical and external quality control isolates, comprising nontuberculous mycobacteria (NTM) (n = 95) and the Mycobacterium tuberculosis complex (MTC) (n = 60), were included in the study. Results: Identification by MBT ML4.0 was correctly performed in 100% of MTC and in 91% of NTM isolates. All of the MTC isolates were correctly differentiated from NTM isolates. Conclusion: Based on our results, MBT ML4.0 may be used reliably to identify both NTM and MTC. © TÜBİTAK.Item Diversity of leishmania strains isolated from cutaneous leishmaniasis patients in Turkey and its reflection to clinics in mice model; [Türkiye’de kutanöz leyşmanyazis hastalarından elde edilen leishmania İzolatlarındaki farklılıklar ve bunların fare modeline klinik yansıması](Ankara Microbiology Society, 2021) Özbilgin A.; Çulha G.; Güray M.Z.; Zeyrek F.Y.; Akyar I.; Töz S.; Östan Ural İ.; Kurt Ö.; Kocagöz T.; Çavuş I.; Gündüz C.Although asexual reproduction has been attributed to Leishmania species, genetic exchange has recently been demonstrated, which helped emerging of hybrid isolates. Situated on the crossroads between three continents, Leishmania hybrids may be present in Turkey. In Turkey, visceral leishmaniasis caused by Leishmania infantum is less common, while cutaneous leishmaniasis (CL) caused by Leishmania tropica and L.infantum could reach 2500 reported cases a year. Our aim was to investigate genetic variability of local Leishmania species and presence of hybrid Leishmania strains in Turkey. Twenty CL patients from Sanliurfa and Hatay, where only L.tropica and both L.tropica and L.infantum cause CL, respectively, were registered equally. All isolates were assessed with real-time polymerase chain reaction (Rt-PCR), isoenzyme analysis, gene sequencing, two-dimensional gel electrophoresis (2D-PAGE) and MALDI-TOF/TOFMS followed by in vivo analyses on mouse model. Identification of differentially expressed proteins was performed. These proteins were confirmed by sequence analysis. All isolates from Sanliurfa were found to be L.tropica which caused cutaneous infection in mice. However, one of 10 isolates from Hatay was found as Leishmania major which caused cutaneous infection. Five isolates were found as L.tropica with Rt-PCR and gene sequencing, one of which had one different protein from the reference L.tropica strain and caused cutaneous infection. Four of the five isolates had five different proteins compared to reference strain and caused both cutaneous and visceral infections. Remaining four isolates showed double melting curves in Rt-PCR, which were concordant with L.tropica and L.infantum. Their sequencing and isoenzyme analyses indicated them as L.infantum. They had six different proteins compared to reference L.infantum strain and caused cutaneous and visceral infections. It is concluded that the isolates with different proteins were hybrid Leishmania species. In the present study, outcomes of the proteomics, genomics, clinical manifestations and tissue tropism on animal models were evaluated together for the first time. In addition to L.tropica and L.infantum, L.major was identified as a causative agent for CL and hybrids of L.infantum/tropica were also shown to be present. © 2020 Ankara Microbiology Society. All rights reserved.Item Determination of antimony resistance mechanism of leishmania tropica causing cutaneous leishmaniasis in Turkey; [Türkiye’de kutanöz leyşmanyazis etkeni leishmania tropica’da antimon direnç mekanizmasının belirlenmesi](Ankara Microbiology Society, 2021) Özbilgin A.; Zeyrek F.Y.; Güray M.Z.; Çulha G.; Akyar I.; Harman M.; Özbel Y.; Ertabaklar H.; Çavuş İ.; Gündüz C.World Health Organization reported that approximately one billion people are at risk in endemic areas, one million cases of cutaneous leishmaniasis (CL) and approximately 300,000 cases of visceral leishmaniasis (VL) were reported per year in the last five years. The number of deaths due to VL is reported to be approximately 20,000 per year. Approximately 2500 cases/year have been reported as CL, caused by Leishmania tropica and Leishmania infantum, in Turkey. The significant increase observed in many cities mainly in the provinces of Mediterranean and Aegean regions in cases and foci in recent years, suggests that there may be an increase in this infections in the following years as well. In Turkey, the causative agent of CL is L.tropica and meglumine antimoniate is used in the treatment of CL. We aimed to determine antimony resistance genes specific for L.tropica by comparing the gene and protein expressions of antimony-resistant and non-resistant L.tropica strains. L.tropica isolates obtained from 3 CL patients without antimonate resistance from Aegean, Mediterranean and Southeastern regions of Turkey were provided to transform into 3 resistant isolates against meglumine antimony in the laboratory conditions. Gene expression alterations by microarray method; protein profiles by two-dimensional gel electrophoresis (2D-PAGE) and relevant proteins by MALDI-TOF/TOF MS of these isolates were accomplished and compared. L.tropica isolates from 10 CL patients who did not respond to antimony therapy were analyzed for resistance to antimonial compounds and quantitative real-time polymerase chain reaction was performed to detect the expression of genes responsible for resistance development. Moreover, differences in protein expression levels in isolates with and without antimony resistance were determined by comparing protein profiles and identification of proteins with different expression levels was carried out. Enolase, elongation factor-2, heat shock protein 70, tripanthione reductase, protein kinase C and metallo-peptidase proteins have been shown to play roles in L.tropica isolates developing resistance to antimonial compounds and similar expression changes have also been demonstrated in naturally resistant isolates from patients. In conclusion, it was revealed that L.tropica strains in our country may gain resistance to meglumine antimoniate in a short time. It is foreseen that if the patients living in our country or entering the country are treated inadequately and incompletely, there may be new, resistant leishmaniasis foci that may increase the number of resistant strains and cases rapidly. © 2020 Ankara Microbiology Society. All rights reserved.Item Progressive Multifocal Leukoencephalopathy Among Ibrutinib Treatment In Chronic Lymphocytic Leukemia(SAGE Publications Ltd, 2022) Çetintepe T.; Gediz F.; Akyar I.; Çetintepe L.; Koç A.M.Introduction: Both chronic lymphocytic leukemia (CLL) itself and the drugs used for its treatment, pose a risk for progressive multifocal leukoencephalopathy (PML). Although the relationship between Rituximab and PML is well known, case reports that have been recently published, suggest that ibrutinib; which is used in the treatment of CLL, may increase the risk of PML. Case report: Here, we report a case of 64 year-old female patient with CLL who was previously treated with rituximab, fludarabine and bendamustin but developed PML after receiving monotherapy with ibrutinib. According to Naranjo’s algorithm, the causality relationship with the drug is possible with a score of 3. The patient initially exhibited neurological symptoms. Magnetic resonance of the brain revealed a bilateral asymmetric hyperintensity in the white matter involving the parietal and occipital lobules, and there was no mass effect, edema, hemorrhagic or iscemic lesions. No enhancement of contrast media was observed. The findings were consistent with demyelination and suggestive of PML. Management and outcome: Mirtazapine treatment was initiated. However, neurological sympthoms continuously progressed over the following weeks and the patient, aged 64, died six weeks after diagnosis of PML. Discussion: PML is a rare and often fatal demyelinating disease of the central nervous system (CNS) that is exclusively seen in immunocompromised patients and there is no specific agent to treat PML. The case discussed here, highlights that the use of ibrutinib in chronic lymphocytic leukemia (CLL) therapy may result in PML. © The Author(s) 2022.