Browsing by Author "Usta, Ayse"

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Determination of Blood Serum Protein Fractions of Calves With Clinically Diagnosed Pneumonia
(Agricultural Research Communication Centre, 2021) Ekici, Pinar Tanritanir; Dede, Semiha; Yuksek, Veysel; Cetin, Sedat; Usta, Ayse
Background: Respiratory diseases in calves are causing worldwide economic losses for the beef industry. This study was planned to determine the serum protein fractions and A/G ratio in calves with clinically diagnosed pneumonia and evaluate the possibility uses of these parameters as clinical diagnostic parameters. Methods: The 34 calves with respiratory system problems and 10 healthy calves without clinical pneumonia symptoms were used as materials. The obtained serum samples were used for total protein and serum protein electrophoresis. Result: Albumin, alpha 1-globulin percentages and A/G ratio decreased in the patient group (p <= 0.01), other globulin fractions were higher in the patient group (p <= 0.01), for % g values. According to concentration results, it was found that while albumin did not show a significant difference, alpha 1-globulin (p <= 0.05) and A/G ratio (p <= 0.01) decreased in the patient group. In addition, other globulin fractions and total protein levels were higher in patient groups (p <= 0.01). As a result, the serum protein fractions should be evaluated as a useful biochemical blood parameter for diagnosis and follow up of lung diseases especially in calves.
Dna Damage-Induced by Sodium Flouride (Naf) and the Effect of Cholicalciferol
(Tech Science Press, 2020) Yuksek, Veysel; Dede, Semiha; Usta, Ayse; Cetin, Sedat; Taspinar, Mehmet
It is known that the high electronegativity of fluorine affects various soft tissues, especially the bone structure in organisms. Of these tissues are the kidneys, which play an important role in the excretion of fluoride from the body. Fluoride affects many cellular mechanisms. One of these effects is DNA damage. Our study aimed to investigate the likely protective effect of cholecalciferol (vitamin D3) on genomic DNA damage-induced NaF depending on concentration and time. The IC25 and IC50 values of NaF for 3, 12 and 24 h and optimum dose of increase in proliferation to vitamin D-3 through MTT assay in NRK-52E kidney cells were determined. DNA damage was significantly increased (p < 0.05) compared to the control group in all groups except for vitamin D-3 It was determined that treatment with NaF together with vitamin D-3 decreased the DNA damage compared to NaF treated groups for 3 and 12 h. NaF combined with vitamin D3 was determined statistically to decrease (p < 0.05) DNA damage compared to NaF treated groups for 24 h. As a result, it was determined that the treatment with cytotoxic concentration NaF depending on the time significantly increased (p < 0.05) the genomic DNA damage, but NaF treatment together with vitamin D-3 decreased the DNA damage in renal cells depending on the time. It was concluded that vitamin D-3 may be useful in preventing DNA damage caused by NaF.
Effect of Imidocarb on Dna Damage in Sheep With Babesiosis
(Kafkas Univ, veteriner Fakultesi dergisi, 2022) Oner, Ahmet Cihat; Ayan, Adnan; Orunc Kilinc, Ozlem; Usta, Ayse; Ertas, Fatma
In this study, it was aimed to determine the DNA damage using the comet assay, which specifically shows DNA damage in naturally Babesia spp.-infected sheep and to evaluate the damage before and after imidocarb application. Blood samples obtained from 10 infected sheep with positive clinical signs and symptoms of babesiosis and whose diagnosis was confirmed by Giemsa staining and PCR methods, and blood samples from 10 healthy sheep were used as study material. DNA damage was examined by the comet assay from the blood samples of the infected patient group and the control group obtained during the disease and after the treatment, and the results were compared with statistical methods. When DNA damage was examined in sick animals diagnosed with babesiosis, the tail length and the tail moment values were found to be statistically significantly higher than the control group (P<0.0001). According to the results obtained after imidocarb application, it was determined that DNA damage and tail moment decreased statistically with imidocarb, and the difference was statistically significant, and the values were higher than the control group (P<0.0001). As a result, Babesia infection can cause DNA damage, has been confirmed by the determination of direct DNA damage using the comet assay, and imidocarb given for treatment was successful and reduced the damage.
The Effect of Lycopene on Dna Damage and Repair in Fluoride-Treated Nrk-52e Cell Line
(Springernature, 2021) Cetin, Sedat; Usta, Ayse; Yuksek, Veysel
Exposure of fluorine at toxic concentrations causes serious damage by accumulating in especially bones, kidneys, and other soft tissues. Fluorine at cytotoxic concentrations may cause DNA damage. This study aims to determine the level of DNA damage due to sodium fluoride (NaF) at different hours (3rd, 12th, and 24th hours) and in IC(50)concentrations designated for each hour and reveal the protective effect of lycopene on possible damage. The best enhancer concentrations (1 mu M) of microtitration (MTT) viability test and proliferation of lycopene and IC(50)values of NaF at the 3rd, 12th, and 24th hour were 9600, 5500, and 3200 mu M, respectively. DNA damage significantly increased in all NaF-treated groups in comparison with the control group (p < 0.05). DNA damage due to NaF+LYC application significantly decreased in comparison with the control group (p < 0.05). Lycopene application significantly increased the expression levels of the Ku70 and Ku80 genes which have a part in DNA repair (p < 0.05). The statistical data showed that application of lycopene which is an important antioxidant molecule may be beneficial for decreasing NaF-induced DNA damage. In conclusion, applying lycopene for cytotoxicity due to fluorine in NRK-52E cell line had different effects based on the dosage and time; thus, it can be a potential option for preventing fluorosis-induced toxicity and developing new treatment approaches.
The Effect of Quinoa (Chenopodium Quinoa) on Apoptotic, Autophagic, Antioxidant and Inflammation Markers in Glucocorticoid-Induced Insulin Resistance in Rats
(Springer, 2022) Erfidan, Siber; Dede, Semiha; Usta, Ayse; Yuksek, Veysel; Cetin, Sedat
Background Insulin resistance plays an important role in predicting type 2 diabetes that may develops. This study was planned in order to investigate the beneficial effects of quinoa (Chenopodium quinoa) use in glucocorticoid induced-insulin resistance. Methods and results Forty-two rats were used as the material (experimental) groups: the control group (C), the quinoa-administered group (Q), the insulin resistance-created group (IR), the IR + metformin group (IM), the IR + quinoa for treatment group (IQ) and the quinoa + IR for prophylaxis group (QI). Blood glucose, insulin levels and HOMA-IR were found to be highest (p < 0.05) in the IR group (p < 0.05). Glucose levels decreased significantly with the administration of quinoa and approached the levels of the control, but the insulin levels and the HOMA-IR did not significantly change. It was also observed that other biochemical parameters (ALT, AST, ALP, total cholesterol, total protein, urea and creatinine) changed significantly in the IR group and approached the levels of the control group with the administration of quinoa. Apoptotic (BCL2 5, BAX 9, CAS 3), autophagic (SQSTM1 7, ATG5) and inflammation (IL-1 beta, TNF-alpha) genes were upregulated by 5-11-fold in the IR group. In the groups in which quinoa was administered for treatment and protection, all these genes were found to be upregulated to a lower extent than the IR group. Antioxidant genes (GPX1, SOD1) increased by nine to tenfold in the quinoa groups. Conclusion As a result, after administration of quinoa, it was determined that the glucose level increased due to experimental insulin resistance and the liver and kidney damage indicators decreased. It was determined that quinoa (Chenopodium quinoa) had significant beneficial effects on biochemical parameters and apoptotic, autophagic, antioxidant and inflammatory markers in experimental glucocorticoid-induced insulin resistance.
Effect of Thymoquinone on Endoplasmic Reticulum (Er) Stress in Nrk-52e Cells
(Univ Karachi, 2023) Celebi, Muhammed; Dede, Semiha; Usta, Ayse
Thymoquinone (TQ), the active component of Nigella sativa, has many beneficial effects. The endoplasmic reticulum involved in the quality control of protein translocation and folding can vary under different conditions, the phenomenon of causing the accumulation of unfolded or misfolded proteins within the ER lumen is termed ER stress. This in vitro study was planned to investigate the effect of TQ on ER stress at proliferative (Tp) and toxic (TQIC50) concentrations on NRK-52E cells at 24th, 48th hours. The expression of important genes in the ER stress pathway (ATF4, ATF6, BIP, CHOP, IRE1, XBP1, PERK) was analyzed. Expression of all genes except CHOP and XBPI increased at 24 hours and BIP at 48 hours for Tp. In the IC50, the CHOP and XBPI gene expressions increased at the 24th hour, and the CHOP and ATF4 genes increased at the 48th hour. As a result, it was determined that the expression of ER stress genes had significant changes with the TQ induction, depending on time and concentration, especially in the proliferative concentration. It is thought that TQ may have varying effects on healthy kidney cells, and it is important to investigate the mechanism of this effect in further studies.
The Effect of Thymoquinone on Nuclear Factor Kappa B Levels and Oxidative Dna Damage on Experimental Diabetic Rats
(Sage Publications india Pvt Ltd, 2017) Usta, Ayse; Dede, Semiha
Background: Thymoquinone (TQ), the basic bioactive phytochemical constituent of seed oil of Nigella sativa, is one of these herbal drugs known for antidiabetic effects. This study was carried out to assess the effects of the possible role of TQ on nuclear factor kappa B (NF-kappa B) and oxidative DNA damage levels in experimental diabetic rats. Materials and Methods: Twenty-eight male Wistar Albino rats (200-250 g) were used as experimental subjects. The rats were divided into four groups, including the control, control supplemented with TQ (CT), diabetic (D), and diabetic supplemented with TQ (DT), each containing seven rats. The D and the DT groups were treated with 45 mg/kg streptozotocin (STZ) (intraperitoneal). TQ was administered 30 mg/kg/day for 21 days by oral gavage in the DT and the T groups. Results: It was determined that glucose, glycosylated hemoglobin (HbA1c) levels and alanine aminotransferase, aspartate aminotransferase, and gamma-glutamyl transpeptidase activities were decreased significantly and approached the control group in the DT group after TQ supplement (P < 0.05). Urea levels were the lowest in CT (P < 0.05). Oxidative DNA damage (8 hydroxy-2-deoxyguanosine) was increased in both of the diabetic groups (D and DT). The NF-kappa B levels were the highest in Group D (P < 0.05). Conclusion: It was observed that increased glucose and HbA1c levels and the indicators of liver and kidney damages were decreased significantly after TQ supplementation. Oxidative DNA damage and NF-kappa B levels were increased in the diabetic group, and TQ administration caused a statistically insignificant reduction.
The Effect of Vitamin E and Selenium Combination in Repairing Fluoride-Induced Dna Damage To Nrk-52e Cells
(Springer, 2020) Yuksek, Veysel; Cetin, Sedat; Usta, Ayse
Prolonged and excessive fluoride exposure can lead to fluorosis. The kidney is one of the organs that are injured mostly due to fluoride-induced damage. Fluoride can induce DNA damage at cytotoxic concentrations. This study aims to determine the extent of NaF-induced DNA damage and to investigate the effect of vitamin E and selenium combination (ES) in preventing and repairing this damage. For this purpose, we administered different combinations of NaF and ES to NRK-52E cells and determined the effective concentrations of ES and the NaF IC(50)values associated with different incubation times (3, 12, and 24 h) by using the MTT assay. The determined quantities of NaF IC(50)in association with time and the NaF IC50+ ES combination were administered to the cells. The extent of DNA damage was determined with the comet assay and the expression levels of the Ku70/80 and PARP-1 genes were determined with the RT-qPCR method. DNA damage significantly increased in all experimental groups compared to the control group (p < 0.05). It was found out that the NaF and ES combination statistically reduced the DNA damage compared to the damage observed in the NaF-treated groups (p < 0.05). Treatment of the ES combination significantly increased the expressions of Ku70 and Ku80 genes involved in DNA repair (p < 0.05). We concluded that vitamin E and selenium can potentially be effective in the repair of fluoride-induced DNA damage based on the results of this in vitro study. Our results may shed light on the prevention of DNA damage associated with fluorosis.
The Effects of Vitamin D Application on Naf-Induced Cytotoxicity in Osteoblast Cells (Hfob 1.19)
(Springernature, 2023) Dede, Semiha; Taspinar, Mehmet; Yuksek, Veysel; Cetin, Sedat; Usta, Ayse
This study was planned to evaluate the effect of vitamin D administration on cytotoxicity due to fluoride exposure in vitro. NaF (IC50) and vitamin D (proliferative) were applied to human osteoblast (hFOB 1.19) cells. The major genes of apoptotic, autophagic, and necrotic pathways were determined by RT-PCR. 2-Delta Delta Ct formulation was used for expression analysis. In the NaF group, caspase 3, Bax, Bad, Bak, Bclx, Atg3, Atg5, Atg6, pG2, LC3-I, LC3-II, RIP1, and RIP3 genes were increased (2.6-15 times). It was observed that the expressions of these genes approached the control when vitamin D was given together with NaF. The Bcl2 gene increased significantly (sixfold) with the effect of NaF, and was down-regulated to some extent with additional vitamin D administration, but still more than in the control. As a result, it was determined that apoptotic, necrotic, and autophagic pathways were activated as the molecular basis of the damage in the bone tissue, which was most affected by fluorine, and these genes were down-regulated and approached the control group with the addition of vitamin D. It was concluded that this is an important data to explain the molecular basis of the protective and therapeutic effect of vitamin D against fluorine toxicity.
In Vitro Evaluation of Thymoquinone and Lycopene Supplementation on Oxidative Dna Damage and Oxidant Status in High Glucose Conditions
(Colegio Farmaceuticos Provincia de Buenos Aires, 2019) Dede, Semiha; Yur, Fatmagul; Taspinar, Mehmet; Cetin, Sedat; Usta, Ayse; Yuksek, Veysel
The present study was planned to investigate the effects of thymoquinone (TQ) and lycopene (LYC), known to possess pro-inflammatory and antioxidant properties, on oxidative DNA damage (8-hydroxy-2-cleoxyguanosine) in BHK-21 cell line treated with high glucose (FIG) and the antioxidant system. BHK-21 cell line was cultured with regular passages (5% FBS, 10% host serum, 1% L-glutamine, 1% penicillin/streptomycin - RPMI 1640, 5% CO2 and 95%, 37 degrees C incubation). MTT cell viability tests were conducted. Proliferative TQ and LYC and glucose IC50 values were determined. Control, study groups; glucose (285 mM), TQ (10 mu M), and LYC (50 mu M)) and cross groups were designed. After incubation, trypsinized cells were broken by the freeze/thaw method and analyzed. Oxidative DNA damage, TAS, TOS and OSI values were determined for the obtained samples. It was determined that 8-OHdG levels were affected by high glucose (p <= 0.05), they increased further with the administration of TQ and LYC in addition to HG. TOS and OSI values increased in all study groups when compared to the control (p <= 0.05), and TAS levels significantly decreased (p <= 0.05) with the administration of HG when compared to TQ and LYC groups. In conclusion, TQ and LYC administration in addition to high glucose exacerbated oxidative DNA damage and OSI, and decreased TAS when compared to TQ and LYC groups. The TQ and LYC dose and administration duration in addition to high glucose in the present study led to an improvement in oxidative balance in the BHK cell line.
Lycopene Prevents Cell Death in Nrk-52e Cells by Inhibition of High Glucose-Activated Dna Damage and Apoptotic, Autophagic, and Necrotic Pathways
(Wiley, 2024) Usta, Ayse; Yuksek, Veysel; Cetin, Sedat; Dede, Semiha
This study aims to investigate the effects of lycopene on apoptotic, autophagic, and necrotic pathways, oxidative status, and DNA damage in diabetic nephropathy at the molecular level. The sample of the study includes seven groups: lycopene (L), high glucose (G), high glucose + lycopene (GL), and control (C) groups tested at 12 and 24 h. The expression levels of genes in oxidative, apoptotic, autophagic, and necrotic cell death pathways are determined by reverse transcription-quantitative polymerase chain reaction analysis. The comet assay method is used for the analysis of DNA damage. It is observed that adding lycopene to high glucose for protective purposes reduces the expression of genes related to apoptosis, autophagy, and necrosis, as well as the DNA damage index, compared to cells given high glucose alone. Lycopene can be a safe and effective alternative agent. In this study, where the molecular mechanisms were examined, it was concluded that lycopene, used as an agent of necrosis, autophagy and apoptosis inhibition, had beneficial effects on cells treated with high glucose. image
Phenolic Contents, Antioxidant Activities, Lcms Profiles of Mespilus Germanica Leaf Extract and Effects on Mrna Transcription Levels of Apoptotic, Autophagic, and Necrotic Genes in Mcf7 and A549 Cancer Cell Lines
(Humana Press inc, 2024) Gormez, Gul; Yuksek, Veysel; Usta, Ayse; Dede, Semiha; Gumus, Selcuk
Cancer, defined by the continuous, uncontrollable proliferation of cells in the human body, is a disease with a rapidly increasing incidence and mortality rate. Scientists are looking for novel ways to cure and prevent this sneaky disease because of the toxicity of contemporary chemotherapy and the cancer cells' resilience to anticancer drugs. Determining the effect of herbal medicines, which do not have as harmful side effects as synthetic drugs, on cancer cell lines is an essential preliminary study in the production of effective drugs against cancer. In this study, the phenolic acid profile, antioxidant capacity, and cytotoxicity of the medicinal plant Mespilus germanica (MG) leaf extract were determined, and its effects on the expression of some apoptotic, necrotic, and autophagic pathway genes of MCF7 (Human breast cancer line) and A549 (Human lung cancer line) and healthy HDF (Human Dermal Fibroblasts) cells were investigated for the first time. The LCMS device detected many important phenolic compounds previously reported to act against cancer cells in Mespilus germanica leaf extract. DPPH and total phenolic content showed high antioxidant capacity. The cytotoxicity of MG was determined by the MTT method. The levels of mRNA transcription for Atg5, Atg3, R & imath;pk1, Bcl2, Bax, Apaf1, Caspase-8, Caspase-7, Caspase-3, and Caspase-9, as well as the expression patterns of the DNA damage markers P53 and Parp-1 genes, were assessed. MG leaf extract did not cause significant toxicity against healthy HDF cells. However, it had a cytotoxic effect on A549 and MCF7 cancer cell lines, increasing the transcription levels of essential genes involved in cell death mechanisms. This research is the first to analyze the phenolic components and antioxidant capabilities of leaf extracts from Mespilus germanica. Additionally, it investigates the impact of these extracts on crucial genes involved in cell death pathways of A549 lung cancer, MCF7 breast cancer, and non-cancerous HDF (Human Dermal Fibroblasts) cells.
Therapeutic Potential of Boric Acid and Borax: Dietary Approaches for Cancer Prevention
(Parlar Scientific Publications (p S P), 2017) Oto, Gokhan; Yildirim, Serkan; Dede, Semiha; Ozdemir, Hulya; Yener, Zabit; Usta, Ayse; Taspinar, Mehmet
In this study, cytoprotective effects of boric acid (BA) and borax (BX) therapy in the rats administered with environmental carcinogens (3-methylcolantren (3-MC) and Benzo(a)pyrene (B(a)P)) were studied. The rats were divided into 9 groups with 12 rats each. Group I was set as control. Group II was administered B(a)P and Group III was administered 3-MC at doses of 100mg/kg intraperitoneally, twice a week, at 4 equal doses. Group IV received only 300 mg/L of BA and Group V was only administered 300 mg/L of BX via drinking water. Group VI received B(a)P + BA, Group VII received 3-MC + BA, Group VIII received B(a)P + BX, and Group IX received 3-MC + BX, and the study was terminated on day 150. In histopathological analysis, hydropic degeneration in liver was seen in the B(a)P group, interstitial pneumonia in lung was seen in the B(a)P + BA group, and hydropic degeneration in liver and interstitial pneumonia in lung were observed in the B(a)P + BX group. Fibrosarcoma occurred in all the groups administered with 3-MC. Fibrosarcoma and diffuse hepatocellular carcinoma developed in the 3 MC group. The administration of BA and BX therapies led to pathological recovery compared to groups administered with B(a)P and 3-MC. This recovery was more prominent in the groups administered with BA therapy. The results of this study indicate that the degree of cytotoxic effect of B(a)P and tumor formations caused by 3-MC may be attenuated by BA and BX therapies.
Vitamin D May Assist the Upr Against Sodium Fluoride-Induced Damage by Reducing Ripk1, Atg5, Becn1, Oxidative Stress and Increasing Caspase-3 in the Osteoblast Mc3t3-E1 Cell Line
(Elsevier Gmbh, 2023) Yuksek, Veysel; Dede, Semiha; Cetin, Sedat; Usta, Ayse; Taspinar, Mehmet
Background: Out of all measure systemic exposure to fluorides can cause defect of skeletal and dental fluorosis. Endoplasmic reticulum (ER) stress is caused by fluorine-induced oxidative stress and importance of vitamin D in its prevention is not known enough in bone cells. This study was carried out to investigate fluorine-induced oxidative stress, ER stress, and death pathways and the effect of vitamin D on them.Methods: MC3T3-E1 mouse osteoblast cell line was used as the material of the study. The NaF and vitamin D concentrations were determined by the MTT assay. NaF treatments and vitamin D supplementation (pre-add, co-add, and post-add) was administered in the cell line at 24th and 48th hours. The expression of the genes in oxidative stress, ER stress, and death pathways was determined using RT-qPCR and Western blotting techniques.Results: Vitamin D significantly reduced mRNA expression levels of SOD2, CYGB, ATF6, PERK, IRE1, ATG5 and BECN1 whereas caused an increase in levels GPX1, SOD1, NOS2 and Caspase-3 in MC3T3-E1 mouse osteoblast cell line of NaF-induced. In addition, GPX1, SOD1, ATF6, PERK, IRE1, BECN1, Caspase-3 and RIPK1 protein levels were examined by Western blot analysis, and it was determined that vitamin D decreased IRE1 and PERK protein levels, but increased GPX1, SOD1, ATF6 and Caspase-3 protein levels.Conclusion: The findings of the study suggest that vitamin D has protective potential against NaF-induced cytotoxicity reasonably through the attenuation of oxidative stress, ER stress, ATG5, IRE1 and by increasesing caspase-3 in vitro conditions.