Browsing by Author "Cakir, Mustafa"

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Alantolactone Ameliorates Graft Versus Host Disease in Mice
(Elsevier, 2024) Odabas, Gul Pelin; Aslan, Kubra; Suna, Pinar Alisan; Kendirli, Perihan Kader; Erdem, Serife; Cakir, Mustafa; Unal, Ekrem
The anti-inflammatory and immunosuppressive drugs which are used in the treatment of Graft-versus-Host Disease (GVHD) have limited effects in controlling the severity of the disease. In this study, we aimed to investigate the prophylactic effect of Alantolactone (ALT) in a murine model of experimental GVHD. The study included 4 BALB/c groups as hosts: Naive (n = 7), Control GVHD (n = 16), ALT-GVHD (n = 16), and Syngeneic transplantation (n = 10). Busulfan (20 mg/kg/day) for 4 days followed by cyclophosphamide (100 mg/kg/day) were administered for conditioning. Allogeneic transplantation was performed with cells collected from mismatched female C57BL/6, and GVHD development was monitored by histological and flow cytometric assays. Additionally, liver biopsies were taken from GVHD patient volunteers between ages 2-18 (n = 4) and non-GVHD patients between ages 2-50 (n = 5) and cultured ex vivo with ALT, and the supernatants were used for ELISA. ALT significantly ameliorated histopathological scores of the GVHD and improved GVHD clinical scores. CD8+ T cells were shown to be reduced after ALT treatment. More importantly, ALT treatment skewed T cells to a more naive phenotype (CD62L+ CD44-). ALT did not alter Treg cell number or frequency. ALT treatment appears to suppress myeloid cell lineage (CD11c+). Consistent with reduced myeloid lineage, liver and small intestine levels of GM-CSF were reduced in ALT-treated mice. IL-6 gene expression was significantly reduced in the intestinal tissue. Ex vivo ALT-treated liver biopsy samples from GVHD patients showed a trend of decrease in proinflammatory cytokines but there was no statistical significance. Collectively, the data indicated that ALT may have immunomodulatory actions in a preclinical murine GVHD model.
A Benzimidazolium Salt Induces Apoptosis and Arrests Cells at Sub-G1 Phase in Epithelial Ovarian Cancer Cells
(Springer, 2024) Akar, Sakine; Cakir, Mustafa; Ozkol, Halil; Akkoc, Senem; Ozdem, Berna
BackgroundOvarian cancer, also known as a silent killer, is the deadliest gynecological cancer in women worldwide. Epithelial ovarian cancers constitute the majority of ovarian cancers, and diagnosis can be made in advanced stages, which greatly reduces the likelihood of treatment and lowers the survival rate. For the treatment of epithelial ovarian cancers, the search for synthetic agents as well as agents of natural origin continues. The effects of 1-(2-cyanobenzyl)-3-(4-vinylbenzyl)-1H-benzo[d]imidazole-3-ium chloride (BD), a benzimidazole derivative, were investigated on epithelial ovarian cancer cells.Methods and resultsIn our study, the effects of BD on proliferation, colony formation, cell death by apoptosis and the cell cycle in A2780 and A2780 Adriamycin (ADR) ovarian cancer cell lines were investigated. Proliferation was examined with cell viability analysis, colony formation and apoptosis with Annexin V staining and cell cycle analyses with PI staining, respectively. As a result of the analyses, BD inhibited cell proliferation and colony formation, induced apoptosis and cell death at 48 h in A2780 and A2780 ADR cells at 10.10 and 10.36 mu M concentrations, respectively. In addition, A2780 and A2780ADR cells were arrested in the Sub-G1 phase of the cell cycle.ConclusionsBD suppresses cancer cell progression by showing antiproliferative effects on ovarian cancer cells. Further analyses are required to determine the mechanism of action of this agent and to demonstrate its potential as a suitable candidate for the treatment of epithelial ovarian cancer.
A Benzimidazolium Salt Induces Apoptosis and Arrests Cells at Sub-G1 Phase in Epithelial Ovarian Cancer Cells (Vol 51, 66, 2024)
(Springer, 2024) Akar, Sakine; Cakir, Mustafa; Ozkol, Halil; Akkoc, Senem; Ozdem, Berna
Characterization of Peripheral Blood T Follicular Helper (Tfh) Cells in Patients With Type 1 Gaucher Disease and Carriers
(Academic Press inc Elsevier Science, 2023) Uzen, Ramazan; Bayram, Fahri; Dursun, Huseyin; Kardas, Fatih; Cakir, Mustafa; Cucer, Nurhan; Donmez-Altuntas, Hamiyet
Background: Gaucher disease (GD) is the most common autosomal recessive lipid storage disease. In this study, the changes in TFH cells and IL-4 and IL-21 cytokines in blood samples of GD patients, carriers and healthy volunteers were investigated.Methods: Two pretreatment type 1 GD patients, 20 currently treated type 1 GD patients, 6 carriers, and 27 healthy volunteers were enrolled in the study. TFH cell (CD45RA- CD4+CXCR5+) number, phenotype (PD1, ICOS expression), and cytokine production (IL-21, IL-4) were assessed via flow cytometric assays.Results: No significant differences were found between the groups with respect to the number, frequency and PD1 or ICOS expression of TFH cells between healthy controls, patients and carriers. However, IL-4+ TFH cells were significantly reduced both in percent and number in the treated GD patients compared with healthy controls (p < 0.05). Interestingly, the IL-21+ TFH cell number was increased in treated GD patients. When TFH cells were examined based on CXCR3 expression, the frequency of the PD1+Th17-Th2-like fraction (CXCR3-) was found to be significantly increased in treated GD patients. Conclusion: To our knowledge, this is the first study to assess TFH cells in GD patients, and to show that the production of IL-4 and IL-21 by TFH cells and their subsets may be altered in type 1 GD patients.
Design and Synthesis of Esipt-Based Imidazole Derivatives for Cell Imaging
(Amer Chemical Soc, 2024) Gul, Sergen; Acikgoz, Eda; Cakir, Mustafa; Menges, Nurettin
Excited-state intramolecular proton transfer (ESIPT)-based fluorescent molecules offer several exciting applications and are utilized most frequently as a cell imaging agent. Because of this, four distinct imidazole derivatives with ESIPT emission have been synthesized, and their fluorescence characteristics have been assessed in a variety of settings. Measurements using fluorescence spectroscopy have shown a promising candidate for cell staining, and potential candidate was specifically investigated for cell imaging uses in HT-29, MDA-MB-231, and HaCaT. Cytotoxicity of candidate molecule (1d) was analyzed using HT-29 and HaCaT cell lines, and at a dosage of 160 mu M, HT-29 and HaCaT cell lines showed no signs of important cell toxicity. When spectroscopically measured, compound 1d showed no fluorescence ability in phosphate-buffered saline (PBS) solution. However, after 8 h of incubation in several cell lines, excellent fluorescence characteristics were seen in the green and red filters.
Design, Synthesis, and Applications of Nucleic Acid-Specific Benzoxazole-N, N-Dialkylphenylamines Derivatives for Nucleolus Imaging in the Cells
(Elsevier, 2024) Kuzu, Burak; Acikgoz, Eda; Cakir, Mustafa
Considered the brain of the nucleus, alterations in the structure and function of the nucleolus are linked to numerous cellular functions and, consequently, contribute to several diseases. The identification of nucleolar morphology and activity via novel biomarkers presents new avenues for the development of therapeutic approaches for a variety of human diseases, including cancer, neurodegeneration, and aging. Therefore, specific detection of the nucleolus with fluorescence probes is of critical importance for clinical applications. In the present study, a series of benzoxazole- N,N -dialkylphenylamines derivative compounds were designed and synthesized based on the benzothiazole-based fluorescence probe Thioflavin T (ThT). Among the compounds, BX-3 and BX-16, which carry electron -withdrawing substituents in the benzoxazole ring, were observed to have higher fluorescence emission at wavelengths of 470 and 465 nM, respectively. The general morphology and divisions of the cells were examined under inverted and light microscopes, respectively, and the fluorescence potentials of selected compounds were determined using immunofluorescence microscopy. The fluorescence intensity of molecules and 3D interactive surface plot images of cells were analyzed using ImageJ software. Cell imaging analyses showed that BX-6 and BX-13, like ThT, specifically stain the cell nucleolus. Moreover, molecular docking studies showed that the compounds could identify the RNA -rich nucleolus by binding with high affinity to the guanine region in the RNA structure. The results suggest that the compounds may be an initial route in developing specific biosensor compounds for nucleolus imaging.
Metformin Eliminates Cd133high Prostate Cancer Stem Cells Via Cell Cycle Arrest and Apoptosis
(Kare Publ, 2021) Acikgoz, Eda; Cakir, Mustafa; Guven, Mustafa; Oktem, Gulperi
Objectives: Cancer stem cells (CSCs), a small subpopulation of tumors, are responsible for chemo-radioresistance, metastasis, and cancer recurrence. The main aim of the present study is to investigate the potential effects of metformin on prostate CSCs (PCSCs). Methods: Flow cytometry was used to isolate cells with co-expression of CD133 and CD44. Sorted PCSCs were treated with different concentrations of metformin to determine the effects of metformin on cell viability using MTT assay. The association of cells exposure to metformin with apoptotic cell death and caspase activity, as well as cell cycle, were performed using the Muse Cell Analyzer. Results: In our study, for the first time we demonstrated the anti-cancer effects of metformin on PCSCs. Our results revealed that treatment with metformin reduced cell viability in CD133(high)/CD44(high) cells in a dose- and time-dependent manner. Metformin significantly induced early apoptosis and triggered the activity of several caspases associated with the apoptotic process. Metformin significantly altered the cell cycle distribution in CD133(high)/CD44(high) cells, leading to G0/G1 phase arrest. Conclusion: The results of the study revealed that metformin triggers cell death and apoptosis and modulates cell cycle distribution in CD133(high)/CD44(high) PCSCS. The present study raises the possibility that metformin is a potential anti-cancer agent for targeting PCSCs.
A Newly Synthesized Benzimidazolium Salt: 1-(2 Chloride as a Potential Anticancer Agent for Colon Cancer Treatment, in Vitro Study
(Wiley, 2025) Bitgen, Nazmiye; Cakir, Mustafa; Akkoc, Senem; Donmez-Altuntas, Hamiyet
Colon cancer is one of the most common cancer-related deaths. Drug resistance is one of the biggest challenges in cancer treatment. Numerous pharmacological and biochemical investigations have documented the benzimidazole ring's anticancer, anti-inflammatory, and antioxidant properties. Within the scope of our project, the effect of newly synthesized benzimidazolium salt (BS) on cell proliferation was tested with MTT assay, and its effect on apoptosis and cell cycle was tested with annexin V and PI in the two different colon cancer cell lines (HT-29 and DLD-1). Our study examined the expressions of some genes related to apoptosis and, additionally, caspase activities with the multicaspase kit.BS showed an antiproliferative effect at lower doses in HT-29 colon cancer cells. When HT-29 cells were exposed to a 20 mu M dosage, they showed increased caspase activity and apoptosis compared to DLD-1 cells. HT-29 accumulated in the G2/M phase of the cell cycle, whereas DLD-1 cells accumulated more in the S phase. In HT-29 cells, colony formation was inhibited; however, in DLD-1 cells, this effect was insufficient.Based on the apoptosis-death pathway, BS is expected to have anti-cancer effects. As a result of this work, this chemical was thoroughly examined in two different colon cancer cell lines, and additional, more comprehensive initiatives are being planned in light of the information obtained from this study.
The Number and Activity of Cd3+tcr Vα7.2+cd161+< Cells Are Increased in Children With Acute Rheumatic Fever
(Elsevier Ireland Ltd, 2021) Ozkaya, Mehmet; Baykan, Ali; Cakir, Mustafa; Vural, Cagdas; Sunkak, Suleyman; Unal, Ekrem; Eken, Ahmet
Background: Acute rheumatic fever (ARF) is an autoimmune disease caused by group A beta-hemolytic streptococci (GAS) and may develop into rheumatic heart disease (RHD). The pathogenesis of ARF and RHD involves molecular mimicry and antibody-mediated mechanisms. T cell involvement is described in various stages of the disease. Mucosal associated invariant T (MATT) cells are enriched at the mucosa and are present in the blood and may be activated by GAS. Methods: In this study, we investigated the quantity and activity of CD3(+) TCRV alpha 7.2(+) CD161(+) cells in the active and recovered ARF patients and healthy controls. Twenty newly diagnosed, 20 recovered-ARF children, and 20 healthy controls were enrolled in the study. Peripheral blood (PB) mononuclear cells were isolated by Ficoll-Paque density gradient. CD4(+) CD4(-) subsets of CD3(+) TCRV alpha 7.2(+) CD161(+) cells and IFN-gamma and TNF-alpha production were quantified by Flow cytometry. Results: Acute and recovered ARF patients had significantly elevated the number of CD3(+) TCRV alpha 7.2(+) CD161(+) cells in their PB. Both CD4(+) and CD4(-) subsets were increased. Moreover, total cell numbers were significantly higher in the recovered patients PB compared with active ARE patients. In addition, CD3(+) TCRV alpha 7.2(+) CD161(+) cells in both acute and recovered patients produced significantly more IFN-gamma and TNF-alpha. Non-MALT total CD3(+) T cell, CD4(+) and CD4(-) T cell subsets were also increased in active and recovered ARF patients and they also produced more IFN-gamma and TNF-alpha. Conclusion: Our data reveal that CD3(+) TCRV alpha 7.2(+) CD161(+) cells are elevated and actively producing IFN-gamma and TNF-alpha in acute and recovered ARF patients and may contribute to ARF pathology. (C) 2021 Elsevier B.V. All rights reserved.
The Number and Frequency of Mucosal-Associated Invariant T (Mait), Γδ T, and Innate Lymphoid Cells (Ilcs) Altered in Patients With Type I Gaucher Disease
(Elsevier Science inc, 2025) Uzen, Ramazan; Bayram, Fahri; Dursun, Huseyin; Kardas, Fatih; Cakir, Mustafa; Cucer, Nurhan; Donmez-Altuntas, Hamiyet
Introduction: Gaucher disease (GD) is a rare lysosomal storage disease caused by mutations in the Glucocerebrosidase (GBA) gene. The innate immunopathology of GD beyond macrophage involvement is not well characterized. In the current study, the changes in ILC subsets, gamma delta T and MAIT cells, TNF-alpha and IFN-gamma cytokine levels in the peripheral blood of patients with Type 1 GD and GD carriers were evaluated. Methods: Peripheral blood mononuclear cells obtained from patients and controls were isolated using the Ficoll-Paque gradient method; after surface and intracellular staining, the cells were analyzed on FACSARIA III. Results: Our analyses revealed that CD8+ MAIT cells and CD8+ gamma delta T cells are reduced in the treated patients compared with the carriers. MAIT cell-specific IFN-gamma production and absolute counts of IFN-gamma+ MAIT cells significantly decreased in Type 1 GD patients who received ERT compared with healthy controls, which could be important indicators for the pathogenesis and severity of the disease. Additionally, total ILCs, particularly the ILC1 subset, were reduced in the Type I GD patients receiving ERT compared with healthy controls and the carriers. Conclusion: The changes observed in ILCs, gamma delta T cells, MAIT cells, TNF-alpha and IFN-gamma cytokine levels in both pre-and post-treatment Type 1 GD patients may play a vital role in the pathogenesis of GD.
Oxidative and Chromosomal Dna Damage in Patients With Type I Gaucher Disease and Carriers
(Pergamon-elsevier Science Ltd, 2023) Uzen, Ramazan; Bayram, Fahri; Dursun, Huseyin; Kardas, Fatih; Altin-Celik, Pinar; Cakir, Mustafa; Donmez-Altuntas, Hamiyet
Background and aims: Gaucher disease (GD) is caused by a genetic deficiency of the beta-glucocerebrosidase enzyme which results in the accumulation of glucosylceramide in macrophages. This accumulation may induce oxidative stress, resulting in DNA damage in patients with GD. The aim of this study was to assess plasma 8-hydroxy-2 '-deoxyguanosine (8-OHdG) levels and cytokinesis-block micronucleus cytome (CBMN-cyt) assay parameters in the peripheral blood lymphocytes of patients with GD and carriers, evaluate the possible associ-ations of these values with GD, and determine whether they can be used as potential biomarkers in GD.Methods: This study included 20 patients with type 1 GD, six carriers, and 27 age-and sex-matched healthy controls. CBMN-cyt assay parameters in peripheral blood lymphocytes of the patients with GD, carriers, and controls were evaluated and 8-OHdG levels in their plasma samples were measured.Results: CBMN-cyt assay parameters in patients with GD and carriers were not significantly different when compared with controls (p > 0.05). However, plasma 8-OHdG levels were found to be higher in both patients with GD and carriers than in control subjects (p < 0.01). Conclusions: Oxidative DNA damage may be a useful prognostic tool, whereas the CBMN-cyt assay cannot be used as a predictive biomarker of GD.
Schisandrin B Treatment After Disease Induction Slightly Improved Experimental Autoimmune Encephalomyelitis
(Sage Publications Ltd, 2023) Yetkin, Mehmet Fatih; Erdem, Serife; Azizoglu, Zehra Busra; Demir, Busra Seniz; Acikgoz, Eda; Cakir, Mustafa; Eken, Ahmet
Temporal Overexpression of Il-22 and Reg3γ Differentially Impacts the Severity of Experimental Autoimmune Encephalomyelitis
(Wiley, 2021) Eken, Ahmet; Erdem, Serife; Haliloglu, Yesim; Zehra Okus, Fatma; Cakir, Mustafa; Fatih Yetkin, Mehmet; Canatan, Halit
IL-22 is an alpha-helical cytokine which belongs to the IL-10 family of cytokines. IL-22 is produced by ROR gamma t+ innate and adaptive lymphocytes, including ILC3, gamma delta T, iNKT, Th17 and Th22 cells and some granulocytes. IL-22 receptor is expressed primarily by non-haematopoietic cells. IL-22 is critical for barrier immunity at the mucosal surfaces in the steady state and during infection. Although IL-22 knockout mice were previously shown to develop experimental autoimmune encephalomyelitis (EAE), a murine model of multiple sclerosis (MS), how temporal IL-22 manipulation in adult mice would affect EAE course has not been studied previously. In this study, we overexpressed IL-22 via hydrodynamic gene delivery or blocked it via neutralizing antibodies in C57BL/6 mice to explore the therapeutic impact of IL-22 modulation on the EAE course. IL-22 overexpression significantly decreased EAE scores and demyelination, and reduced infiltration of IFN-gamma+IL-17A+Th17 cells into the central nervous system (CNS). The neutralization of IL-22 did not alter the EAE pathology significantly. We show that IL-22-mediated protection is independent of Reg3 gamma, an epithelial cell-derived antimicrobial peptide induced by IL-22. Thus, overexpression of Reg3 gamma significantly exacerbated EAE scores, demyelination and infiltration of IFN-gamma+IL-17A+ and IL-17A+GM-CSF+Th17 cells to CNS. We also show that Reg3 gamma may inhibit IL-2-mediated STAT5 signalling and impair expansion of Treg cells in vivo and in vitro. Finally, Reg3 gamma overexpression dramatically impacted intestinal microbiota during EAE. Our results provide novel insight into the role of IL-22 and IL-22-induced antimicrobial peptide Reg3 gamma in the pathogenesis of CNS inflammation in a murine model of MS.