Browsing by Author "Oztatlici, M"

Now showing 1 - 7 of 7
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    In-Vitro Evaluation of Immunomodulation Effects of Mesenchymal Stem Cell-Derived Exosomes in Refractory Chronic Spontaneous Urticaria
    Ozdemir, AT; Kirmaz, C; Ozgul Ozdemir, RB; Oztatlici, M; Kilicarslan Sonmez, P; Tuglu, MI
    Objective: Approximately half of chronic spontaneous urticaria (CSU) patients are thought to have an autoimmune pathology, and they are resistant to current treatment approaches. Mesenchymal stem cells (MSCs) are adult cells that have been shown to be useful in many autoimmune pathologies due to their immunomodulation properties. This study aimed to investigate the immunomodulatory effects of MSCs, and exosomes isolated from refractory CSU patients.Materials and Methods: Peripheral blood mononuclear cells (PBMCs) were isolated from 5 refractory CSU patients and 5 healthy volunteers. The effects of MSCs isolated from CSU patients and healthy MSCs were compared. Co-culture experiments were performed to evaluate the efficacy of Mesenchymal Stem Cells and exosomes on PBMCs of CSU patients and healthy volunteers. To compare the resulting effects, changes in IFN-gamma, IL-4, IL-10, IL-17a, and TGF-(3 cytokines were detected by the ELISA method. Cell proliferations were detected with the CCK-8 kit.Results: The effects of autologous and allogeneic MSCs on IFN-gamma expressions were similar, both providing significant suppression at all cell ratios. However, IL-4 and IL-10 expression of PBMCs co-cultured with allogeneic MSCs significantly decreased while IL-17a and TGF-(3 expression increased significantly. In addition, our findings indicated that exosomes were capable of significant suppression at low PBMC ratios, regardless of autologous or allogeneic origin, but MSCs were more effective as the number of PBMCs increased.Conclusion: These preliminary findings from in-vitro experiments suggested that allogeneic MSC, or high-dose exosome administration may be a potential approach for treatment in CSU patients, most of whom are regarded as suffering from an autoimmune disease and resistant to current treatments. However, our findings need to be supported by clinical studies.
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    In-vitro Evaluation of Effects of Mesenchymal Stem Cells on TLR3, TLR7/8 and TLR9-activated Natural Killer Cells
    Ozdemir, AT; Kirmaz, C; Ozdemir, RBO; Degirmenci, P; Oztatlici, M; Degirmenci, M
    Objectives: In this study, it was aimed to investigate the immunomodulatory effects of Mesenchymal stem cells (MSCs) on Natural Killer (NK) cells activated by Toll-like receptor (TLR) agonists. Methods: MDA-MB-231, MCF-7 and NK-92 cells were induced with TLR3, TLR7/8 and TLR9 agonists and co-cultured with MSCs. Alterations in IFN-gamma, TNF-alpha, Granzyme-b and Perforin expressions were determined by qPCR method, CD69 and CD107a expressions were determined by flow cytometry, and cytotoxicity was determined by MTT-assay. Results: All TLR agonists significantly increased the expressions of the IFN-gamma, TNF-alpha, Granzyme-b, Perforin, CD69 and CD107a in-vitro. We determined that the cytokine, cytotoxic molecules, and activation markers of NK-92 cells interacting with breast tumor cells significantly increased by TLR3 and TLR9 agonists. However, suppression rather than activation occurred on the NK-92 cells due to the simultaneous induction of the immunosuppressive effects of MSCs by these agonists. On the other hand, the TLR7/8 agonists provided a low NK-92 induction, however, the inhibitory effects of MSCs were not triggered. Therefore, it provided a more significant activation than TLR3 and TLR9 agonists. Conclusion: Our findings suggested that TLR7/8 agonists may be a better choice to induce antitumor effects of NK cells in a tumor tissue rich in MSCs.
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    Molecular Analysis of Cattle Isolates of Echinococcus granulosus in Manisa Province of Turkey
    Altintas, N; Oztatlici, M; Altintas, N; Unver, A; Sakarya, A
    Echinococcus granulosus is the causative agent of cystic echinococcosis (CE) in humans and many domestic animals, and still one of the most important global health problem in the world and in Turkey. Infection with metacestode causes severe illness and high economic losses. Several strains of Echinococcus have been identified based on the epidemiological and biological characteristics of strains. In this study, a total of 18 individual hydatid cyst samples from cattle were examined. They were obtained from central slaughterhouse in the province of Manisa/Turkey between 2010-2012. The total genomic DNA (gDNA) was extracted using RTA-DNA Isolation Kit (Gebze/Kocaeli, Turkey) according to manufacturer instructions from protoscoleces and cystic germinal membranes. The aim of this study was to provide molecular characterization of E. granulosus isolates which were obtained from cattles by using polymerase chain reaction (PCR) in Manisa province of Turkey. After PCR, to investigate the genetic characteristics of isolates, deoxyribonucleic acid sequencing of the mitochondrial cytochrome c oxidase subunit 1 (CO1) and nicotinamide adenine dinucleotide dehydrogenase subunit 1 (NAD1) genes were performed with ABI Prism Genetic Analyzer 3100 instrument. As a result of our study, all (18) cattle isolates were detected as E. granulosus sensu stricto (G1-G3 complex). This is the first molecular study report genotyping of Echinococcus isolates from cattle in Manisa province.
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    Placental expressions of Anti-Mullerian hormone/Receptor, vascular endothelial growth factor and related microRNAs in patients with preeclampsia: a case control study
    Hasdemir, PS; Celikcekic, D; Oztatlici, M; Ozbilgin, K
    Anti-Mullerian hormone (AMH) has been implicated in the pathogenesis of preeclampsia. The present study was primarily designed to determine the placental tissue AMH, Anti-Mullerian hormone Receptor II (AMHRII), vascular endothelial growth factor (VEGF) and microRNA (miRNA) 26a/126/155/210 expressions and serum miRNA 26a/126/155/210 levels in patients with preeclampsia to examine their potential role in the pathogenesis of preeclampsia. Placental tissue samples from patients with preeclampsia (n = 20) and control subjects (n = 20) were examined by immunohistochemical staining and quantitative polymerase chain reaction (qPCR) for AMH, AMHRII, VEGF mRNA expression levels and miRNA 26a/126/155/210 expressions. Serum levels of miRNA 26a/126/155/210 were measured by qPCR. Patients with preeclampsia had lower AMH/AMHRII immunostaining, particularly in syncytiotrophoblastic cells compared to control subjects (p < 0.05). The relative mRNA expressions of AMH/AMHRII were increased (1.535 +/- 0.121 and 1.155 +/- 0.049 fold, p < 0.0002 and p < 0.033, respectively) and the relative mRNA expression of VEGF was decreased (4.878 +/- 0.331 fold, p < 0.0002) in patients with preeclampsia compared to control subjects. The miR-26a expression was increased and miR-126 expression was decreased in serum samples of patients with preeclampsia compared to control subjects (p < 0.0002). miR-155 and miR-210 expressions were increased in serum and placental tissue samples of patients with preeclampsia compared to control subjects (p < 0.0002). In conclusion, reduced placental tissue immunostaining of AMH/AMHRII along with increased AMH/AMHRII mRNA expressions may indicate posttranscriptional dysregulation. Robust increase in expressions of hypoxia/inflammation-related miRNAs particularly miR-155 and miR-210 might have a role in this mechanistic pathway. Increased serum levels of miR 26a, 155 and 210 are potential early diagnostic markers for preeclampsia.
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    Cyclophosphamide suppresses spermatogenesis in the testis of mice through downregulation of miR-34b and miR-34c
    Özbilgin, MK; Demirören, S; Üçöz, M; Oztatlici, M
    Cyclophosphamide (CP) is commonly used as an anticancer agent but has been associated with high toxicity in several organs, including the testes. In this study, we aimed to evaluate the effects of CP-induced testicular toxicity, using glial cell line-derived neurotrophic factor (GDNF), occludin and transforming growth factor beta 3 (TGF-beta 3) primary antibodies, and miR-34b and miR-34c expressions. Eighteen young Balb/c male mice were divided into three groups. The control group received no treatment. The mice of CP group were injected 100 mg kg(-1) day(-1) CP for 5 days, and the same amount of saline was injected in the sham group. The animals were sacrificed 24 hr after the last injection. Immunohistochemical analysis of testicular tissues showed a decrease in both spermatogenic germ cell count and also GDNF, occludin expressions, but an increase in TGF-beta 3 expression in the CP group compared to the others group. The expressions of miR-34b and miR-34c were examined by qPCR technique, a significant decrease was observed in tissue samples in the CP-treated group. The expression of GDNF, occludin and TGF-beta 3 plays an important role in testicular injury caused by CP, and the decrease in the expression of miR-34b/c in tissue samples may be an important marker for the detection of testicular damage.
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    Immunomodulatory Effects of MDA-MB-231-derived Exosome Mimetic Nanovesicles on CD4+ T Cell Line
    Oztatlici, M; Ozdemir, AT; Oztatlici, H; Kucukhuyuk, S; Ozdemir, RBO; Orhan, H; Ozbilgin, K
    Objectives: The aim of this study is to investigate the immunomodulatory effects of MDA-MB-231 cells or MDA-MB231-derived exosome-mimetic nanovesicles (NVs) on CD4+ Jurkat cells. Methods: NVs were produced by the breakdown of MDA-MB-231 cells and the characterization of generated NVs were performed by using direct-ELISA and Flow cytometry methods. We co -cultured CD4+ Jurkat cells with MDA-MB-231 cells or MDA-MB-231-derived NVs for 48 h. Subsequently, expressions of pro -inflammatory and anti-inflammatory cytokines, and related transcription factors of CD4+ Jurkat cells were evaluated by qPCR method. Results: Clustering, which is the indicator of activation, was not seen in the CD4+ Jurkat cells co -cultured with MDAMB-231 cells. However, CD4+ Jurkat cell clusters were observed in the co -culture experiments with all NV concentrations. In addition, it was determined that the expressions of pro -inflammatory cytokines significantly increased while the expressions of anti-inflammatory cytokines was dramatically decreased in the NV -treated groups. On the contrary, opposite results were obtained in CD4+ Jurkat cells co -cultured with MDA-MB-231 cells. Moreover, TNF-alpha and Gata3 expressions were decreased in all groups. Conclusion: These preliminary findings from in -vitro experiments suggested that NVs could be a potential tool in cancer immunotherapy, but our data need to be supported by more comprehensive studies.
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    Histological and electroencephalographic demonstration of probiotic effect for reduce of oxidative stress and apoptosis in experimental traumatic brain injury
    Karakayali, EM; Kocamaz, E; Alpay, S; Onal, T; Oztatlici, M; Durusma, R; Ozel, HF; Mete, M; Barutcuoglu, M; Kutlu, N; Tuglu, MI
    BACKGROUND: The gut microbiota modulates nervous system function. In the literature, it has been shown that this modula-tion is used in many nervous system injuries through oxidative stress (OS) and apoptosis mechanisms. In this study, it was aimed to investigate the neuroprotective effects of probiotic (PB) treatment in a rat traumatic brain injury (TBI) model with histological and electroencephalographic (EEG) data.METHODS: Forty male Wistar albino rats were divided into four groups. Group 1 was the control group (CONTROL, n=10) and no trauma was applied. Group 2 was the trauma group with the weight-drop technique (TBH, n=10). Group 3 was the sham group (SHAM), (TBH+sterile saline [SS], n=10) rats were given 500 mu L of SS per day by oral gavage. Group 4 was the PB treatment group, (TBH+PB, n=10) rats were treated daily for 7 days with 500 mu L of PB oral gavage. Brain samples were collected 7 days after trauma. Histopathological evaluation of brain samples was done with HE. OS with Endothelial nitric oxide synthase, vascularization with Vascular Endothelial Growth Factor, gliosis with S100, and apoptosis with caspase 3 were evaluated immunohistochemically. Apoptotic index was determined with TUNEL. In addition, EEG and somatosensory evoked potential (SEP) recording findings were compared.RESULTS: It was determined by HE staining that there was a significant (P<0.001) damage in the TBI and sham groups compared to the control group. It was found that PB treatment provided a significant (P<0.01) improvement in the damage created. While OS (P<0.01), gliosis (P<0.01), and apoptosis (P<0.05) decreased with PB treatment, angiogenesis (P<0.01) increased. In support of these findings, in the software-mediated EEG and SUP examination; Delta wave power and theta/alpha ratio increased with TBI and de-creased with PB treatment.CONCLUSION: The results showed that PB treatment provided a significant improvement in rats by reducing OS, apoptosis, and gliosis and increasing vascularity. To the best of our knowledge in the literature, it was shown for the 1st time that histological results for the treatment of PB were supported by software-mediated EEG and SEP analysis.