Browsing by Author "Taspinar, Mehmet"

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The Antagonistic Effects of Temozolomide and Trichostatin a Combination on Mgmt and Dna Mismatch Repair Pathways in Glioblastoma
(Humana Press inc, 2023) Guven, Mustafa; Taspinar, Filiz; Denizler-Ebiri, Farika Nur; Castresana, Javier S.; Taspinar, Mehmet
Glioblastoma is the most aggressive and fatal form of brain cancer. Despite new advancements in treatment, the desired outcomes have not been achieved. Temozolomide (TMZ) is the first-choice treatment for the last two decades and has improved survival rates. Emerging studies have shown that targeting epigenetics in glioblastoma can be beneficial when combined with clinically used treatments. Trichostatin A (TSA), a histone deacetylase inhibitor, has anti-cancer properties in various cancers. No data concerning the TMZ and TSA relationship was shown previously in glioblastoma therefore, we aimed to determine the likely therapeutic effect of the TMZ and TSA combination in glioblastoma. The T98G and U-373 MG, glioblastoma cell lines, were used in this study. TMZ and TSA cytotoxicity and combination index were performed by MTT assay. The expression of DNA repair genes (MGMT, MLH-1, PMS2, MSH2 and MSH6) was detected using RT-PCR. One-way analysis of variance (ANOVA) was used for statistical analysis. Combination index calculations revealed antagonistic effects of TMZ and TSA in terms of cytotoxicity. Antagonistic effects were more apparent in the T98G cell line, which is expressing MGMT relatively higher. MGMT and DNA Mismatch Repair (MMR) genes were upregulated in the T98G cell line, whereas downregulated in the U373-MG cell lines under TMZ and TSA combination treatment. It is concluded that MGMT might be playing a more active part than MMR genes in TMZ resistance to TMZ and TSA antagonism. This is the first study elucidating the TMZ and TSA relationship in cancer cell lines.
Caveolin-1 Polymorphisms in Patients With Severe Obstructive Sleep Apnea
(Taylor & Francis Ltd, 2017) Asker, Selvi; Taspinar, Mehmet; Koyun, Hasan; Ozbay, Bulent; Arisoy, Ahmet
Objective: To investigate the associations of G14713A and T29107A polymorphic variants of Caveolin-1 with severe obstructive sleep apnea (OSA). Materials and methods: This study was performed on 86 severe OSA patients and 86 controls. Genotyping was performed to investigate the association of G14713A and T29107A polymorphisms of Caveolin-1 with severe OSA. Results: The distribution of genotypes of T29107A was significantly different between controls and OSA patients with a higher proportion of TT carriers in the OSA group. Conclusion: T29107A-specific genotype of Caveolin-1 may be linked with severe OSA pathogenesis.
Chemotherapeutic Resistance in Anaplastic Astrocytoma Cell Lines Treated With a Temozolomide-Lomeguatrib Combination
(Springer, 2014) Ugur, Hasan Caglar; Taspinar, Mehmet; Ilgaz, Seda; Sert, Fatma; Canpinar, Hande; Rey, Juan A.; Sunguroglu, Asuman
The treatment of anaplastic astrocytoma (AA) is controversial. New chemotherapeutic approaches are needed for AA treatment. Temozolomide (TMZ) is one of the chemotherapeutic drugs for the treatment of AA. The cytotoxic effects of TMZ can be removed by the MGMT (O(6)-methylguanine-DNA methyltransferase) enzyme. Then, chemotherapeutic resistance to TMZ occurs. MGMT inhibition by MGMT inactivators (such as lomeguatrib) is an important anticancer therapeutic approach to circumvent TMZ resistance. We aim to investigate the effect of TMZ-lomeguatrib combination on MGMT expression and TMZ sensitivity of SW1783 and GOS-3 AA cell lines. The sensitivity of SW1783 and GOS-3 cell lines to TMZ and to the combination of TMZ and lomeguatrib was determined by a cytotoxicity assay. MGMT methylation was detected by MS-PCR. MGMT and p53 expression were investigated by real-time PCR after drug treatment, and the proportion of apoptotic cells was analyzed by flow cytometry. When the combination of TMZ-lomeguatrib (50 mu M) was used in AA cell lines, IC50 values were reduced compared to only using TMZ. MGMT expression was decreased, p53 expression was increased, and the proportion of apoptotic cells was induced in both cell lines. The lomeguatrib-TMZ combination did not have any effect on the cell cycle and caused apoptosis by increasing p53 expression and decreasing MGMT expression. Our study is a pilot study investigating a new therapeutic approach for AA treatment, but further research is needed.
Determining the Relationship Between Atherosclerosis and Periodontopathogenic Microorganisms in Chronic Periodontitis Patients
(Taylor & Francis Ltd, 2017) Bozoglan, Alihan; Ertugrul, Abdullah Seckin; Taspinar, Mehmet; Yuzbasioglu, Betul
Objectives: The aim of this study is to determine the relationship between atherosclerosis and periodontopathogenic microorganisms in chronic periodontitis patients following periodontal treatment. Materials and Methods: A total of 40 patients were included in the study. 20 of these patients diagnosed with atherosclerosis and chronic periodontitis formed the test group. The remaining 20 patients were systemically healthy patients diagnosed with chronic periodontitis and formed the control group. All patients had nonsurgical periodontal treatment. The periodontopathogenic microorganism levels were determined at baseline and at 6 months in microbial dental plaque samples and WBC, LDL, HDL, PLT, fibrinogen, creatinine and hs-CRP levels were determined by blood samples. Results: Statistically significant reduction has been achieved in clinical periodontal parameters following non-surgical periodontal treatment in test and control groups. Following periodontal treatment, WBC, LDL, PLT, fibrinogen, creatinine and hs-CRP levels significantly decreased and HDL levels significantly increased in both test and control groups. Similarly, the periodontopathogenic microorganism levels significantly decreased following periodontal treatment in the test and control groups. A statistically significant positive correlation has been determined between the periodontopathogenic microorganism levels and WBC, LDL, PLT, fibrinogen, creatinine, and hs-CRP levels in the test group. Conclusions: The association between hs-CRP, WBC, LDL, PLT, fibrinogen, creatinine, and the amount of periodontopathogenic microorganisms indicates the possibility that periodontal treatment could decrease the risk atherosclerosis. More studies must be conducted in order for these results to be supported.
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.
An Easy Synthetic Protocol for Imidazo-1,4 and Evaluation of Their Toxicities
(Wiley-hindawi, 2018) Kuzu, Burak; Genc, Hasan; Taspinar, Mehmet; Tan, Meltem; Menges, Nurettin
Imidazo-1,5-alkynyl alcohol derivatives were synthesized, and they were cyclized to imidazo-1,4-oxazines by means of cesium carbonate. Propargyl-allene isomerization was examined, and the reaction mechanism was proposed. Moreover, cytotoxicity of synthesized molecules against LN405 cell lines was investigated by means of structure-activity relationship (SAR). With SAR study, toxicities of some functional groups have been shown. In addition, two lead compounds were tested against DNA damaging.
The Effects of Some Minerals on Apoptosis and Dna Damage in Sodium Fluoride-Administered Renal and Osteoblast Cell Lines
(int Soc Fluoride Research, 2019) Cetin, Sedat; Yur, Fatmagul; Taspinar, Mehmet; Yuksek, Veysel
The present study was planned to investigate the effects of some minerals (MgCl2, Na2SeO3, AlCl3 CaCl2) on the expression and translocation of certain apoptotic markers in NaF-administered (at the rate of IC50) rat renal epithelial (NRK-52E) and human osteoblast (hFOB 1.19) cell lines. The NaF IC50 and the non-toxic mineral doses were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. For the biochemical analysis, the cells were collected by trypsin treatment following a 24-hour incubation period, the cells were broken up by the freeze/thaw method, and the analysis was conducted. Caspase 9, 8, and 3 levels and gene expression, and M30 and 8-OHdG levels were determined. Target gene DNAs were propagated with the real time-PCR method. In the MTT studies, it was determined that the cell proliferation in rat renal epithelial cells (NRK-52E) treated with NaF+minerals was higher than that of all the NaF-treated groups and in the human osteoblast cells (hFOB 1.19), the cell proliferation was higher than in all the NaF-treated groups except for the MgCl2 group. The reason why the NaF administration in the NRK-52E cells resulted in an average of a 2-fold decrease in caspase 3 expression compared to the control group could be attributed to the apoptotic effect of NaF based on the time we obtained the RNA. However, based on this time, when the results are assessed based on the NaF and other mineral groups, the NaF-induced cytotoxic apoptosis might have used a pathway other than the apoptotic pathway. Thus, it is considered that minerals could usually prevent NaF-induced apoptosis by a synergistic mechanism due to the ionic character of NaF. NaF+mineral administration protected the NRK-52E cells from apoptosis. In the osteoblasts, on the other hand, it was concluded that NaF+mineral administration may be useful since it inhibits increased apoptosis.
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.
The Effects of Vitamins A, D, E, and C on Apoptosis and Dna Damage in Sodium Fluoride-Treated Renal and Osteoblast Cell Lines
(int Soc Fluoride Research, 2017) Yuksek, Veysel; Dede, Semiha; Taspinar, Mehmet; Cetin, Sedat
This study was planned to investigate the effects of the antioxidant and protective vitamins A, D, E, and C, on the expression and translation of certain apoptotic markers in the NRK-52E and hFOB 1.19 cell lines treated with NaF at half the maximal inhibitory concentration (IC50) for 24 hours. The IC50 for NaF and nontoxic vitamin doses were determined by the MTT viability test. For the biochemical assays, cells were harvested by trypsinization and lysed by the freeze/thaw method. The levels and gene expression of caspases 3, 8, and 9, and the levels of M30 and 8-OHdG were also measured with methods that included the use of ELISA and qRT-PCR. In the MTT studies, compared to the NaF-treated groups, it was found that the cell viability was higher in all the NaF+vitamin D-treated groups in the NRK-52E cell line, in some of the NaF+vitamin D-treated groups in the hFOB 1.19 cell line, and in some of the NaF+vitamin A, E, and C-treated groups for both cell lines. In the NRK-52E cell line, the NaF IC50 value was determined and found not to induce apoptosis sufficiently so that it was considered that mechanisms other than the apoptotic pathways were instrumental in causing cell death. In the hFOB 1.19 cell line, it was observed that the apoptotic M30 protein level was increased in the NaF+vitamin D and NaF+vitamin C groups. In addition, in the hFOB 1.19 cell line, the qRT-PCR results showed that, while the expression of caspase-3 increased with vitamin A and that of caspase-8 increased with NaF, treatment with NaF+vitamin A led to a lower levels of caspases 3 and 8. Future studies to investigate the most valid and active mechanism for NaF-induced cell death and to elucidate the inhibitory-activating effects of vitamins on this mechanism using different doses, durations of exposure, and analytic methods should be considered.
Exploring the Combined Anti-Cancer Effects of Sodium Butyrate and Celastrol in Glioblastoma Cell Lines: a Novel Therapeutic Approach
(Humana Press inc, 2024) Kartal, Bahar; Denizler Ebiri, Farika Nur; Gueven, Mustafa; Taspinar, Filiz; Canpinar, Hande; Cetin, Sedat; Taspinar, Mehmet
Glioblastoma, a highly aggressive and lethal brain cancer, lacks effective treatment options and has a poor prognosis. In our study, we explored the potential anti-cancer effects of sodium butyrate (SB) and celastrol (CEL) in two glioblastoma cell lines. SB, a histone deacetylase inhibitor, and CEL, derived from the tripterygium wilfordii plant, act as mTOR and proteasome inhibitors. Both can cross the blood-brain barrier, and they exhibit chemo- and radiosensitive properties in various cancer models. GB cell lines LN-405 and T98G were treated with SB and CEL. Cell viability was assessed by MTT assay and IC50 values were obtained. Gene expression of DNA repair, apoptosis, and autophagy-related genes was analyzed by RT-PCR. Cell cycle distribution was determined using flow cytometry. Viability assays using MTT assay revealed IC50 values of 26 mM and 22.7 mM for SB and 6.77 mu M, and 9.11 mu M for CEL in LN-405 and T98G cells, respectively. Furthermore, we examined the expression levels of DNA repair genes (MGMT, MLH-1, MSH-2, MSH-6), apoptosis genes (caspase-3, caspase-8, caspase-9), and an autophagy gene (ATG-6) using real-time polymerase chain reaction. Additionally, flow cytometry analysis revealed alterations in cell cycle distribution following treatment with SB, CEL and their combination. These findings indicate that SB and CEL may act through multiple mechanisms, including DNA repair inhibition, apoptosis induction, and autophagy modulation, to exert their anti-cancer effects in glioblastoma cells. This is the first study providing novel insights into the potential therapeutic effects of SB and CEL in glioblastoma.
Glutatyon Uygulamasının Yüksek Glukoz Ortamında Oksidatif Dna Hasarı ve Antioksidan Sistem Üzerine Etkilerinin İn Vitro Olarak Değerlendirilmesi
(2022) Taspinar, Mehmet; Yur, Fatmagul; Dede, Semiha; Çetın, Sedat; Usta, Ayşe
Amaç: Bu çalışma, yüksek oranda glukoz ilave edilen BHK-21 hücre serisinde antioksidan özellikleri bilinen glutatyonun hücrelerde olası oksidatif DNA hasarı (8-hidroksi-2-deoksiguanozin) ve antioksidan sistem üzerine etkilerini ortaya koymak amacıyla planlandı. Gereç ve Yöntem: Bu amaçla, BHK-21 hücre serisinde in vitro koşullarda düzenli pasajları yapılarak (%5 FBS, %10 horse serum, %1 L-Glutamin, %1 penisilin/ streptomisin içeren RPMI 1640 besi yerinde ve %5 CO2 ve % 95 nem ve 37oC’de) inkübe edildi. MTT hücre canlılık testleri yapılarak glutatyonun kontrol grubuna göre ve glukozun IC50 değeri belirlendi. Hücreler pleytlere, 2x106 hücre olacak şeklide ekildi. Kontrol ve deneme grupları ve bu gruplar arasında çaprazlama olarak, çalışma grupları (glukoz; (285 mM), glutatyon (250 μM) hazırlandı. Yirmi dört saatlik inkübasyonu takiben tripsine edilen hücreler, dondur/çöz yöntemiyle parçalanarak analize hazırlandı. Elde edilen hücre kültürü lizatında; oksidatif DNA hasarı, TAS, TSO ve OSİ değerleri ELISA ile spektrofotometrik olarak ölçüldü. Bulgular: 8-OHdG düzeyleri, glukoz uygulanması ile önemli oranda arttığı, HG+GSH grubunda kontrole göre anlamlı olarak arttığı saptandı (p≤0.05). Sadece GSH verilen grup ise kontrolden farksız olarak bulundu. TAS bakımından, GSH uygulanan gruplarda kontrole fark bulunmazken, HG +GSH verilen grupda ise kontrole göre önemli artış tespit edildi (p≤0.05). TOS ve OSİ ise HG+GSH uygulanan gurupda kontrole göre önemli bir artış gözlendi (p≤0.05). Öneri: Çalışmadan elde edilen sonuçlara göre yüksek glukoz uygulanan hücrelere uygulanan glutatyonun hücresel düzeyde bu dozlarda koruyucu etkisi gözlenmemiştir. Ancak glutatyon uygulanan guruplarda, uygulanan glutatyon dozlarının hücreler üzerine toksik doz olmadığı da belirlenmiştir.
Hhv-6 Is Ubiquitously Found Using Western Blot in Tonsils and Adenoid Tissues of Healthy People
(Edizioni int Srl, 2013) Taspinar, Mehmet; Cetin, Nilgun; Gerceker, Devran; Karasartova, Djursun; Turegun, Buse; Ozturk, Sibel; Sahin, Fikret
Few studies have examined the prevalence and cellular proclivity of latent human herpesyirus 6 (HHV-6) in healthy populations. Difficulties in detection of HHV-6 genome in different tissues using polymerase chain reaction (PCR) and immunohistochemistry (IHC) techniques have been reported by various researchers. We examined tonsils and adenoid tissues of 54 patients who had undergone tonsillectomy or adenoidectomy without any evidence of acute infection for the presence of latent HHV-6 infection. While we were investigating the prevalence of HHV-6, we tested the efficiency of PCR, IHC and Western Blot (WB) for detection of HHV-6 in tonsil tissues. We found that 100% of tonsil tissues were positive for HHV-6 with WB, 40% of tonsils were positive with PCR and no tonsil was positive with IHC. This result correlates well with most studies claiming HHV-6 is a ubiquitous organism in various populations and tissues. Western blot may be a good choice for detecting HHV-6 in tissues. Expression of the HHV-6 gp60/110 envelope protein disclosed by WE may indicate that HHV-6 does not have true latency. To our knowledge, this is the first report to use WB to test for HHV-6 in tissues.
In Vitro Evaluation of the Effects of Lycopene on Caspase System and Oxidative Dna Damage in High-Glucose Condition
(Wolters Kluwer Medknow Publications, 2019) Bazyel, Bunyamin; Dede, Semiha; Cetin, Sedat; Yuksek, Veysel; Taspinar, Mehmet
Aim and Background: The present study was planned to investigate the effects of lycopene, on the caspase-dependent apoptosis in high-dose glucose (HG)-treated PC12 cell line. PC12 cells were cultured in vitro. Materials and Methods: HG was prepared as G (250 mM), and lycopene was prepared as L1 (10 mM), L2 (20 mM), and L3 (40 mM). After 6 h of incubation, the cells were exposed to trypsin, and the samples were obtained with freeze/thaw method. Caspase 3, 8, 9; 8-hydroxy-2-deoxyguanosine (8-OHdG); and M30 were determined (enzyme-linked immunosorbent assay). Results: 8-OHdG increased in L3 (P % 0.001), whereas L1 caused a decrease in HG group (P % 0.001). Caspase-3 decreased significantly in L1, L2, and L3G compared to control (P % 0.001) group. Caspase-8 increased significantly in L1, L1G, L2G, and all L3 glucose groups (P % 0.001). There was no difference for Caspase-9. M30 was not affected by L and HG, which decreased significantly (P % 0.001). Conclusion: As a result, it was determined that, when PC12 cell line was treated with HG, lycopene application had effects on caspase enzymes and DNA damage.
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.
In Vivo Evaluation of Thymoquinone on Apoptosis and Oxidative Dna Damage in High Glucose Condition
(C M B Assoc, 2018) Gumus, Ali Furkan; Dede, Semiha; Yuksek, Veysel; Cetin, Sedat; Taspinar, Mehmet
The study was planned to investigate the effects of thymoquinone (TQ), which is a compound in N. sativa, on caspase dependent apoptosis and oxidative DNA damage in high glucose treated PC12 cells. PC12 cells were treated with high glucose (G1-150 mM, G2-250 mM, G3-350 mM), TQ (20 mu M), and their combinations. Oxidative DNA damage (8-OHdG (8-Oxo-2'-deoxyguanosine)), and apoptosis (caspase 3, caspase 8, caspase 9 enzymes and M30 protein) parameters were analyzed with ELISA. The 8-OHdG levels decreased in all combination groups compared to the control (p <= 0.001). There was no statistically significant difference between caspase 3 and 9. Caspase 8 in TQ, G3, TQG1, TQG2 groups were higher than the control (p <= 0.002). Low M30 levels were observed in TQG1 group (p <= 0.002). In conclusion, it was observed that in PC12 cell line treated with the high glucose concentrations, TQ administration had a statistically significant effect on oxidative DNA damage and some apoptotic parameters (caspase 8 and M30 protein).
Investigation of Genetic Structure in Van Cats Using Microsatellite Markers
(Elsevier Science Bv, 2017) Koyun, Hasan; Koncagul, Seyrani; Karakus, Kadir; Okut, Hayrettin; Kucuk, Mursel; Yilmaz, Ayhan; Taspinar, Mehmet
Mono- or Di-Substituted Imidazole Derivatives for Inhibition of Acetylcholine and Butyrylcholine Esterases
(Academic Press inc Elsevier Science, 2019) Kuzu, Burak; Tan, Meltem; Taslimi, Parham; Gulcin, Ilhami; Taspinar, Mehmet; Menges, Nurettin
Mono- or di-substituted imidazole derivatives were synthesized using a one-pot, two-step strategy. All imidazole derivatives were tested for AChE and BChE inhibition and showed nanomolar activity similar to that of the test compound donepezil and higher than that of tacrine. Structure activity relationship studies, docking studies to on X-ray crystal structure of AChE with PDB code 1B41, and adsorption, distribution, metabolism, and excretion (ADME) predictions were performed. The synthesized core skeleton was bound to important regions of the active site of AChE such as the peripheral anionic site (PAS), oxyanion hole (OH), and anionic subsite (AS). Selectivity of the reported test compounds was calculated and enzyme kinetic studies revealed that they behave as competitive inhibitors, while two of the test compounds showed noncompetitive inhibitory behavior. ADME predictions revealed that the synthesized molecules might pass through the blood brain barrier and intestinal epithelial barrier and circulate freely in the blood stream without binding to human serum albumin. While the toxicity of one compound on the WS1 (skin fibroblast) cell line was 1790 mu M, its toxicity on the SH-SY5Y (neuroblastoma) cell line was 950 mu M.
A Novel Polymerase Chain Reaction To Detect Brucella Canis in Dogs
(Kafkas Univ, veteriner Fakultesi dergisi, 2015) Aras, Zeki; Taspinar, Mehmet; Aydin, Ibrahim
In this study, the specific polymerase chain reaction has been standardized and evaluated for the direct diagnosis of Brucella canis in vaginal swab samples from dogs. The specific primer sets are directed to the 16S-23S rRNA inter-space region of Brucella spp. and the deletion of 351 bp in BMEI1426-BMEI1427 in B. canis. A total of 21 references and field strains and 35 vaginal swab samples were used for the evaluation of the polymerase chain reaction. It found that polymerase chain reaction is positive for B. canis DNA indicated by only amplification of 214 bp product. It detected at least 2.7 x 10(1) CFU/g of bacteria diluted in vaginal swab samples indicates that the polymerase chain reaction can be used as a practical alternative for bacterial isolation. The novel polymerase chain reaction provides a simple and rapid for the detection of B. canis in clinical and field samples in one step and in short time about 24 h.
Raffinose and Hypotaurine Improve the Post-Thawed Merino Ram Sperm Parameters
(Academic Press inc Elsevier Science, 2013) Bucak, Mustafa Numan; Keskin, Nazan; Taspinar, Mehmet; Coyan, Kenan; Baspinar, Nuri; Cenariu, Mihai C.; Kursunlu, Ahmet Nuri
The aim of this study was to determine the effects of raffinose and hypotaurine on sperm parameters after the freeze-thawing of Merino ram sperm. Totally 40 ejaculates of five Merino ram were used in the study. Semen samples, which were diluted with a Tris-based extender containing 10 mM raffinose, 5 mM hypotaurine, 5 mM raffinose +2.5 mM hypotaurine (H + R) and no antioxidant (control), were cooled to 5 degrees C and frozen in 0.25 ml French straws and stored in liquid nitrogen. Frozen straws were then thawed individually at 37 degrees C for 25 s in a water bath for evaluation. The addition of raffinose led to higher percentages of subjective and CASA motilities (47.5 +/- 12.2%, 46.3 +/- 13.6%) compared to controls (38.8 +/- 13.8%, 30.5 +/- 11.7%, P < 0.05). For the CASA progressive motility, 5 mM raffinose (20.12 +/- 8.82%) had increasing effect in comparison to control (10 +/- 7.94%, P <0.05) following the freeze-thawing process. Raffinose and hypotaurine led to higher viability (40.8 +/- 4.68%, 40.8 +/- 4.7%), high sperm mitochondrial activity (29.5 +/- 5.4%, 27.3 +/- 4.9%) and acrosome integrity (50.8 +/- 8.1, 50.7 +/- 4.4) percentages, compared to control groups (31.5 +/- 3.5%, 9.5 +/- 8.2%, 42.8 +/- 7.3%, P < 0.05). H + R group only led to high sperm mitochondrial activity when compared to control group. In the comet test, raffinose and hypotaurine resulted in lower sperm with damaged DNA (6.2% and 3.9%) than that of control (9.1%), reducing the DNA damage. For TUNEL assay, The TUNEL-positive cell was distinguished by distinct nuclear staining. Raffinose and H + R groups resulted in lower sperm with TUNEL-positive cell (1.5 +/- 1.2% and 2.1 +/- 0.9%) than that of control (4.9 +/- 2.5%) (P < 0.05). In conclusion, findings of this study showed that raffinose and hypotaurine supplementation in semen extenders provided a better protection of sperm parameters against cryopreservation injury, in comparison to the control groups. Crown Copyright (C) 2013 Published by Elsevier Inc. All rights reserved.
The Role of Hbd-2, Hbd-3, and Calprotectin in the Relationship Between Chronic Periodontitis and Atherosclerosis
(Tech Science Press, 2020) Taspinar, Mehmet; Bozoglan, Alihan; Ertugrul, Abdullah Seckin; Elmas, Levent
This study was carried out to compare individuals diagnosed with atherosclerosis and periodontal periodontitis based on the degree of change in the human beta-defensins (HBD) HBD-2, HBD-3, and calprotectin. Atherosclerosis is the most frequently observed cardiovascular disease. Dental and periodontal infections are known to provide a considerable basis for atheroma plaque formation. The study group consists of a total number of 40 subjects, with 20 patients diagnosed with atherosclerosis and chronic periodontitis and 20 systemically healthy patients diagnosed with chronic periodontitis. Clinical periodontal and blood parameters and HBD-2, HBD-3, and calprotectin biomarkers in the gingival crevicular fluid were measured. In both groups, following clinical periodontal treatment, a statistically significant decrease in white blood cells (WBC), low-density lipoproteins (LDL), fibrinogen, creatinine, and platelets (PLT), a statistically significant increase in high-density lipoproteins (HDL) in blood samples, statistically meaningful decrease in HBD-2, HBD-3, and calprotectin in the gingival crevicular fluid were achieved. Blood values and HBD-2, HBD-3, calprotectin amounts in the gingival crevicular fluid were increased significantly in the test group compared to the control group. A positive correlation was observed between decreases in HBD-2, HBD-3, calprotectin, and clinical periodontal indices. Regression in systemic inflammation was observed after clinical periodontal treatment. It is concluded that nonsurgical periodontal treatment of chronic periodontitis positively affects atherosclerosis prognosis.