Browsing by Author "Turgay N."
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Item Short report: Quantiferon-Leishmania as an epidemiological tool for evaluating the exposure to Leishmania infection(2010) Turgay N.; Balcioglu I.C.; Toz S.O.; Ozbel Y.; Jones S.L.The aim of the present preliminary study was to investigate the potential of measurement of IFN-γ secretion by T cells into blood plasma using QuantiFERON assay with leishmanial antigens to determine the presence of Leishmania infection. Blood samples from cured visceral (N = 18), and cutaneous (N = 20) leishmaniasis cases, and 20 healthy controls were tested. The IFN-γ responses to Leishmania major H2B and Leishmania infantum H2B antigens were detected from the majority of treated old visceral leishmaniasis cases, but not from controls. Future studies using larger groups will be required to establish the true potential of the assay for epidemiological screening of leishmaniasis. Copyright © 2010 by The American Society of Tropical Medicine and Hygiene.Item The comparison of parasitological and bacteriological stool examinations in patients with gastrointestinal symptoms; [Gastrointestinal sistem yakınması olan hastalarda dışkının parazitolojik ve bakteriyolojik incelemelerinin karşılaştırılması](Veteriner Fakultesi Dergisi, 2012) Bayram A.; Oyur T.; Ünver A.; Aydemir Ş.; Özacar T.; Özensoy Töz S.; Turgay N.Stool samples of 236 patients with acute and chronic gastrointestinal symptoms admitted to Ege University Medical Faculty Parasitology Outpatient Clinic Laboratory between July 2009 to June 2010 were examined. One hundred eleven out of 236 (47%) patients admitted to the laboratory during summer and autumn period with acute gastrointestinal symptoms while other 125 patients (53%) admitted during winter and spring with chronic symptoms. At least one parasite was determined in 112 out of 236 (47.45%) patients. In retrospective analysis, it has determined that the bacteriological examination of stool samples were also performed in 121 out of 236 (51.7%) patients. Seven out of 121 (5.78%) patients admitted both bacteriological and parasitological laboratories together had at least one bacterial agent in their stool examination. Five out of 7 patients were found to be having bacteriological and parasitological mixed infection. Our findings showed that it is important to perform both bacteriological and parasitological examinations together in patients admitting to hospital with intestinal symptoms due to these mixed infections. Performing both diagnostic techniques together will improve accurate diagnosis, treatment and understanding possible etiological reasons of these mixed infections. © 2012, Veteriner Fakultesi Dergisi. All rights reserved.Item E.Histolytica/dispar cases diagnosed in ege university medical faculty parasitology outpatient clinic between January 2010 to June 2011; [Ocak 2010-haziran 2011 tarihleri arasında ege üniversitesi tıp fakültesi parazitoloji polikliniğinde saptanan e. Histolytica/dispar olguları](Veteriner Fakultesi Dergisi, 2012) Ünver A.; Oyur T.; Kurt Ö.; Özensoy Töz S.; Turgay N.For definitive diagnosis of amoebiasis, The Ministry of Health requires the detection of Entamoeba histolytica (E. histolytica) trophozoites that ingested red blood cells in Trichrome-stained smears or E. histolytica-specific adhesin antigen with ELISA. Stool samples of 51 patients admitted to the Outpatients Clinic of Ege University School of Medicine Department of Parasitology between January 2010 and June 2011 were suspected to have E. histolytica/dispar cysts or trophozoites in wet mount examinations and stained with Trichrome. Examination of these smears revealed that 49 samples were positive for E. histolytica/dispar Thirty-three samples were tested for the positivity of E. histolytica-specific adhesin antigen with a commercial ELISA kit (Entamoeba CELISA-Path; CeLLabs Pty. Ltd., Brookvale, Australia) and 23 were found to be positive. Our results indicated an elevation of figures of amoebiasis cases in recent years. It is concluded that application of different methods for the diagnosis of E. histolytica infections as suggested by The Ministry of Health is essential for correct reports of peripheral laboratories. © 2012, Veteriner Fakultesi Dergisi. All rights reserved.Item Genotyping of giardia lamblia in a cohort of turkish patients: A search for a relationship between symptoms and genotypes; [Bir türk hasta kohortunda giardia lamblia genotiplendirilmesi: Semptomlar ile genotipler arası ilişkinin araştırılması](Veteriner Fakultesi Dergisi, 2012) Balcioglu C.; Kurt O.; Sevil N.; Dagci H.; Tetik A.; Ergunay K.; Yereli K.; Ozbilgin A.; Turgay N.; Ozensoy Toz S.Recent surveys investigating the molecular biology of Giardia lamblia revealed two distinct assemblages with different clinical outcomes. However, there is not a universal compromise about the clinical effects of each assemblage, warranting further studies. Here, we report the results of the first analyses of the assemblages of G. lamblia in Manisa province located in western Turkey, together with their relationships with the symptoms and DNA sequence analyses of the PCR products. DNA samples were isolated from the stools of 63 patients infected with G. lamblia and 54 DNA samples, amplified successfully with PCR, were digested with the enzyme Xho I for RFLP. Thirty-eight of 54 samples (70.4%) were found to be in Assemblage A, while the remaining 16 samples (29.6%) were found to be in Assemblage B. The number of female patients was found significantly higher in Assemblage B (P=0.18). There was a statistically significant relationship between the occurrence of both abdominal pain and diarrhea and Assemblage B (chi-square, 10.52; P<0.05). No other statistically significant relationship was detected between the assemblages and neither with the symptoms nor with the age groups of the patients. The comparison of the DNA sequences of the PCR products from two assemblage B (one subtype B1 and one B) and one assemblage A samples both with each other and with other DNA sequences in the NCBI website by multialignment analyses, revealed specific regions for assemblages B (B1-B) and A on tpi gene region. Further studies with more patients are required to assess these initial results. Now, our aim is to design a probe for tpi gene region to set up a real-time PCR assay that is easier to conduct and requiring shorter time for the analyses. © 2012, Veteriner Fakultesi Dergisi. All rights reserved.Item The importance of the contribution of rapid test, serological and molecular methods in the diagnosis of two imported malaria cases with atypical microscopy; [Mikroskopide Atipik Gorunumlu Dis Kaynakli Iki Sitma Olgusunda Hizli Test, Serolojik ve Molekuler Yontemlerin Taniya Katkisinin Onemi](Ankara Microbiology Society, 2017) Zorbozan O.; Pullukcu H.; Sahar E.A.; Karakavuk M.; Can H.; Tunali V.; Doskaya M.; Turgay N.; Toz S.; Ozbilgin A.Malaria is a widespread and life-threatening disease in tropical and subtropical regions. In patients with typical clinical symptoms, malaria is considered as a preliminary diagnosis if there is a travel history to malaria-endemic areas. The basis of the laboratory diagnosis of malaria is the microscopic examination of Giemsa stained smears. On the other hand, the diagnosis and differentiation of Plasmodium species with microscopic examination may have some difficulties. In the first case, adifferent appearance from the classical Plasmodium vivax erythrocytic forms in infected erythrocytes were detected in 1% of all erythrocytes in thin smear blood preparations of a 26-year-old male with complaints of fever and chills and a story of travel to Nigeria. It was observed that parasitic nuclei were not prominent, and were located in the cytoplasm irregularly as chromatin or dye particles, nucleus fragments similar to Schiiffner's granules in the form of scattered and granular spots were present in some erythrocytes, the cytoplasm of some Plasmodium erythrocytic forms were irregular and nuclei were not seen. There were no Schiiffner's granules in any of the infected erythrocytes. PMvax was detected by the rapid diagnostic test (OptiMAL, DiaMed GmbH, Switzerland), which searches for the antigens of Plasmodium species, in the peripheral blood sample of the patients. The P.vlvax 18S rRNA gene was also detected by the multiplex real-time polymerase chain reaction. Antibodies against Plasmodium species were searched by using the Pan Malaria Antibody CELISA (CeLLabs Pty Ltd, Brookvale, Australia) kit in the patient's serum sample and the optical density (OD) value of the patient sample was measured five times the OD value of the positive control. In the second case, adifferent appearance from the classical P.faldparum erythrocytic forms in infected erythrocytes were detected in 12% of all erythrocytes in thin smear blood preparations of a 31-year-old male who has been suffering from persistent fever, severe headache, pain in the eyes and was known to be working in Nigeria. It was observed that some Plasmodium trophozoites have 1 /3 of the size of erythrocytes such as P.vivax and have non-granular cytoplasm, some erythrocytic forms were round and the nucleus and cytoplasm were hardly distinguished, some of them were seen as crescent and close to the nucleus of the cytoplasm and some erythrocytic forms had characteristically a single nucleus and a scattered cytoplasm, similar to mature trophozoites of P.vivax. Although the Plasmodium young trophozoites were similar to Rvtvax in means of magnitude, the forms in which the nude adhered to the erythrocyte wall were common. There were no Rfalciparum gametocyte forms. Rfalciparum like young trophozoite was observedonly in one of the four smears. P.falciparum was detected by the commercial rapid diagnostic test and Rfalciparum 18S rRNA gene was also detected by the multiplex real-time polymerase chain reaction. Antibody formation against Plasmodium species was not detected in the ELISA test. In these case reports, the importance of the support of rapid diagnostic tests, serological and molecular methods to microscopic diagnosis and species determination of two imported malaria cases were demonstrated.Item Infecting glial cells with antimony resistant Leishmania tropica: A new ex-vivo model; [Glia Hücrelerinin Antimona Dirençli Leishmania tropica ile Enfekte Edilmesi: Yeni Bir ex-vivo Modeli](Ankara Microbiology Society, 2018) Zorbozan O.; Harman M.; Evren V.; Erdoǧan M.A.; Kilavuz A.; Tunali V.; Çavuş I.; Yilmaz O.; Özbilgin A.; Turgay N.Leishmaniasis is a vector-borne zoonotic disease that shows different clinical features like cutaneous, mucocutaneous, visceral and viscerotropic forms. The protocols used in the treatment of leishmaniasis are toxic and have many limitations during administration. One of the limitations of treatment is the resistance against the protocols in practice. There is also a need to define new treatment options especially for resistant patients. Ex-vivo models using primary cell cultures may be a good source for evaluating new drug options in patients with antimony resistance, in addition to in-vitro and in-vivo studies. In this study, it was aimed to define a new ex-vivo culture model to evaluate treatment options in patients with cutaneous leishmaniasis who did not respond to treatment. In our experimental model of ex-vivo infection, Leishmania tropica promastigotes isolated from a case previously diagnosed with cutaneous leishmaniasis were used. The primary astroglial cell culture used for the ex-vivo model was prepared from 2-3 days old neonatal Sprague Dawley rat brains under sterile conditions by the modification McCarthy's method. The astroglia cells, which reached sufficient density, were infected with antimony resistant Ltropica promastigotes. After 24 hours of incubation, the supernatant on the cells were collected, the cell culture plate was dried at room temperature, then fixed with methyl alcohol and stained with Giemsa to search for Ltropica amastigotes. Amastigotes were intensely observed in glia cells in primary cell cultures infected with Ltropica promastigotes. No promastigotes were seen on Giemsa stained preparations of the precipitates prepared from the bottom sediment after the centrifugation of the liquid medium removed from the infected plates. In this study, promastigotes from a cutaneous leishmaniasis patient unable to respond to pentavalent antimony therapy were shown to infect rat glia cells and converted to amastigote form. This amastigote glial cell model, as far as we know, is the first model in the literature produced by Ltropica. The occurrence of Ltropica amastigote forms in glia cells may be indicative of the ability of Leishmania species to infect the central nervous system. The central nervous system may be an area for the Leishmania amastigotes to escape from the immune system in cases of leishmaniasis without a treatment response. Our study is important because it is the first study to show the infection of glia cells with L.tropica amastigotes. © 2018 Ankara Microbiology Society. All rights reserved.Item Evaluating the Glucantime Concentration for the ex vivo Glial Cell Model of Antimony-resistant Leishmania tropica Amastigotes; [Antimon Dirençli Leishmania tropica Amastigotlarının ex vivo Glial Hücre Modeli için Glukantim Konsantrasyonunun Değerlendirilmesi](Galenos Publishing House, 2021) Zorbozan O.; Evren V.; Harman M.; Özbilgin A.; Yılmaz Ö.A.; Turgay N.Objective: Because the protocols used in the treatment of leishmaniasis can be toxic and have many limitations, such as the development of resistance against such protocols, new treatment options are needed, especially against resistant patients. Ex vivo models may be a good source for evaluating new drug options for patients with antimony-resistant parasites. This study aimed to evaluate the Glucantime concentration for our ex vivo glial cell amastigote model we had defined in previous work. Methods: We prepared the astroglial cell culture from brains of 2 to 3 day old neonatal Sprague-Dawley rats under sterile conditions by modifying McCarthy’s method. Four plates of cells were infected with antimony-resistant Leishmania tropica promastigotes. After 24 h of incubation, we added Glucantime to 3 plates with different concentrations. After 72 h, we removed the supernatant and then dried, fixed, and stained the plates with Giemsa to count the amastigotes in the glial cells. Results: We observed the amastigotes in glial cells in the control flask. Glial cells were ruined in flasks, which include 75 µg/ mL and 37.5 µg/mL Glucantime. The number of amastigotes per 100 glial cells was 116 for the flask with 7.5 µg/mL Glucantime concentration, while 487 for the control flask. Conclusion: We found that while high concentrations of Glucantime were toxic for glial cells, 7.5 µg/mL Glucantime concentration managed to reduce the number of Leishmania tropica amastigotes in glial cells. © 2021 Turkish Society for Parasitology-Available online at www.turkiyeparazitolderg.orgItem Overcoming the Challenge; In Vivo Efficacy of Miltefosine for Chronic Cutaneous Leishmaniasis(Springer Science and Business Media Deutschland GmbH, 2021) Tunalı V.; Harman M.; Çavuş İ.; Gündüz C.; Özbilgin A.; Turgay N.Background: Cutaneous Leishmaniasis (CL) is the most common form of leishmaniasis. CL can be divided into two major groups: acute CL (ACL) and chronic CL (CCL). The aim of this study is to compare the efficacy of miltefosin and pentavalent antimony compounds in vivo with the CCL patient samples. Materials: Three study groups were formed, each consisting of five male Mus musculus (Balb/C) mice. In this model, promastigotes from the culture of a CCL patient were utilized. 100 μL L. tropica promastigote suspension with a density of 108 promastigotes/ml were injected into the hint-right footpad of each experimental animal intradermally. Footpads of the mice were measured every two weeks until 24th week. From the 13th week, miltefosin 50 mg/kg/day was administered orally using gavage for 21 days, Meglumin antimoniate (MA) was administered by intramuscular (IM) injection daily for 21 days at 50 mg/kg/day and saline was administered IM for 21 days for the miltefosine, MA and control group, respectively. Results: The footpad measurements of the miltefosine group were lower than the control group statistically. Between the MA group and the miltefosine group and MA group and the control group, there was no statistically significant difference. Giemsa stained slides revealed amastigotes in one, two and all of the slides for the miltefosine, MA and control group, respectively. Molecular tests were performed with the Rotor-Gene device and L. tropica consistent peaks were obtained in one of the miltefosine group, four in the MA group and all mice in the control group. Conclusions: Demonstration of both clinical and laboratory improvement in four of the five experimental animals provides strong evidence that miltefosine is an effective drug in the treatment of CCL. In the literature, no clinical or laboratory studies using miltefosine have been performed with CCL patients only. © 2020, Witold Stefański Institute of Parasitology, Polish Academy of Sciences.Item Investigation of in vitro Efficacy of Miltefosine on Chronic Cutaneous Leishmaniasis; [Kronik Kutanöz Leishmaniasis’te Miltefosin Etkinliğinin in vitro Olarak Araştırılması](Galenos Publishing House, 2022) Tunalı V.; Harman M.; Çavuş İ.; Zorbozan O.; Özbilgin A.; Turgay N.Objective: Leishmaniasis is the second deadliest parasitic disease in the World Health Organisation’s list of neglected diseases, following malaria. Cutaneous leishmaniasis (CL) is the most common form of the disease and it is one of the few communicable diseases with increasing incidence rates owing to factors like armed conflicts and climate change. CL can be divided into two major groups: Acute CL (ACL) and chronic CL (CCL). The aim of this study was to compare the in vitro efficacy of miltefosine and pentavalent antimony compounds in the CCL patient samples. Methods: Five isolates previously isolated from 5 CCL patients were included in this study. Genotyping is performed using internal transcribed spacer 1 (ITS 1) gene region real-time PCR. In vitro drug efficacy tests were applied to determine their activity against meglumine antimoniate (MA) and miltefosine. Serial dilutions (512, 256, 128, 64, 32, 16, 8 and 4 µg/mL) prepared from MA and miltefosine were prepared in 96-well flat-bottom cell culture plates and incubated at 24 °C for 48 hours. The efficacy of the drug on Leishmania spp. promastigotes after 24 and 48 hours was evaluated by hemocytometer slide and XTT cell viability test. Results: All of the samples were genotyped as L. tropica. Evaluation of 24 and 48 hours showed, 128 µg/mL and 256 µg/mL and 32 µg/mL and 64 µg/ mL concentrations of miltefosine and MA were enough to kill all the promastigotes respectively. The results of the hemocytometer slide and XTT were consistent. Conclusion: There are no studies investigating the in vitro efficacy of miltefosine with the CCL patient group. To overcome the treatment challenges experienced in this special patient group, more studies are needed. According to our results, it is concluded that miltefosine is efficient for the treatment of CCL and further clinical studies with miltefosine will reveal valuable data. © 2022 Turkish Society for Parasitology.Item Autochthonous transmission of Leishmania donovani and Leishmania major with all the components of infection cycle at Europe's doorstep(Elsevier B.V., 2022) Özbilgin A.; Tunalı V.; Akar Ş.Ş.; Yıldırım A.; Şen S.; Çavuş I.; Zorbozan O.; Gündüz C.; Turgay N.; İnanır I.Objectives: Leishmaniasis is a vector-borne disease and dogs may act as urban reservoirs. Turkey and most of the Mediterranean basin countries are endemic for leishmaniasis. In this study, it is aimed to report the autochthonous leishmaniasis cases, with all the components of the infection cycle (reservoir, vector, and the host) in a region close to Europe. Methods: Nine human and four canine autochthonous leishmaniasis cases were included in the study. Direct microscopy, culture methods, serological, and molecular tests were applied to the samples obtained from the cases. Results: VL and CL patients consisted of 2 L.infantum, 1 L. donovani, 2 L. tropica, and 2 L. tropica,1 L. major,1 L. infantum infected patients respectively. CanL cases were infected with L. infantum, L. donovani, L. tropica, and L. major. Conclusions: All the cases were autochthonous cases located in Manisa province. As Greece and all the Mediterranean basin countries in Europe share competent vectors, it is concluded that the detection of all 4 species of Leishmania parasites in such proximity to Europe poses an important public health threat for Europe. This study reports all four species of Leishmania spp., including L. major and L.donovani in close proximity to continental Europe. © 2022 Elsevier B.V.Item Unpleasant Souvenir: Imported Plasmodium falciparum Malaria in Türkiye; [Nahoş Hatıra: Türkiye’de Yurtdışı Kaynaklı Plasmodium falciparum Sıtması](Galenos Publishing House, 2023) Özbilgin A.; Tunalı V.; Akar Ş.Ş.; Çavuş İ.; Zorbozan O.; Yıldırım A.; Turgay N.Objective: Each year, approximately 125 million people visit malaria-endemic countries. This study aimed to investigate the clinical characteristics of imported Plasmodium falciparum malaria infections in Türkiye. Methods: The study included patients diagnosed with P. falciparum malaria between 1996 and 2022. A retrospective evaluation was conducted on whole blood samples and/or blood smears, as well as detailed medical histories, clinical manifestations, and laboratory findings. A total of 131 imported cases of P. falciparum were included in the study. Results: Among the patients, 121 were male. Of these, 101 had traveled to Africa, while 30 had visited Asia. Among the patients, 109 were returned travelers, and 22 were refugees/migrants. Early trophozoites were observed in all patients, while gametocytes were detected in 30 patients. Cerebral malaria developed in 15 patients, resulting in the death of two individuals. Additionally, 10 patients received preventive chemoprophylaxis. Conclusion: Turkey is situated on migration routes that connect two continents to Europe, where more than 95% of the global malaria burden exists. The importation of malaria through returned travelers poses a risk of malaria reintroduction in our country, given the presence of suitable vectors, climate conditions, and environmental factors. Importantly, 30 patients (22.9%) exhibited gametocyte forms of P. falciparum, which have the potential to infect Anopheles species, thus establishing a basis for local malaria transmission. © 2023 Turkish Society for Parasitology.