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Adipokine, gut and thyroid hormone responses to probiotic application in chukar partridges (Alectoris chukar) exposed to heat stress
(Akademiai Kiado Zrt, 2021) Bayraktar, Bulent; Tekce, Emre; Kaya, Hacer; Gurbuz, Ahmet Burak; Dirican, Ebubekir; Korkmaz, Serol; Atalay, Banu
The aim of this study was to investigate the effect of Lactobacillus reuteri E81 (LRE) probiotic supplementation on heat stress responses in chukar partridges (Alectoris chukar). The birds were divided into two groups, one of which was exposed to heat stress (HS). Within each group, four subgroups, each including 64 birds, were created for the three treatment doses (200, 400 or 600 mg/kg) of LRE and the control. The experiment was started with day-old birds, kept at a temperature of 25 degrees C or 37 degrees C. After a 7-day adjustment period, the LRE supplementation lasted for 35 days. The levels of different adipokines, including visfatin (VF), adiponectin (ADP), chemerin (CHEM), as well as the concentration of plasma citrulline (CIT) and the levels of thyroid hormones (T3 and T4) and thyroid-stimulating hormone (TSH) in the blood were measured at 21 and 42 days of age. A significant correlation (P < 0.01) was found between LRE supplementation and the decrease in serum VF, ADP, CIT, T3 and T4 levels in partridges exposed to HS. On the other hand, no significant relationship was found between LRE supplementation and the serum CHEM and TSH levels (P > 0.05). We concluded that the addition of 600 mg/kg LRE is beneficial in preventing intestinal damage and inflammation provoked by HS.
Analysis of mitochondrial DNA cytochrome-b (CYB) and ATPase-6 gene mutations in COVID-19 patients
(Wiley, 2022) Dirican, Ebubekir; Savrun, Seyda Tuba; Aydin, Ismail Erkan; Gulbay, Gonca; Karaman, Ulku
Coronavirus disease of 2019 (COVID-19) is a pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Mutations of mitochondrial DNA (mtDNA) are becoming increasingly common in various diseases. This study aims to investigate mutations in the cytochrome-b (CYB) and adenosine triphosphatase-6 (ATPase-6) genes of mtDNA in COVID-19 patients. The association between mtDNA mutations and clinical outcomes is investigated here. In the present study, mutations of the mtDNA genes CYB and ATPase-6 were investigated in COVID-19 (+) (n = 65) and COVID-19 (-) patients (n = 65). First, we isolated DNA from the blood samples. After the PCR analyses, the mutations were defined using Sanger DNA sequencing. The age, creatinine, ferritin, and CRP levels of the COVID 19 (+) patients were higher than those of the COVID-19 (-) patients (p = 0.0036, p = 0.0383, p = 0.0305, p < 0.0001, respectively). We also found 16 different mutations in the CYB gene and 14 different mutations in the ATPase-6 gene. The incidences of CYB gene mutations A15326G, T15454C, and C15452A were higher in COVID-19 (+) patients than COVID-19 (-) patients; p < 0.0001: OR (95% CI): 4.966 (2.215-10.89), p = 0.0226, and p = 0.0226, respectively. In contrast, the incidences of A8860G and G9055A ATPase-6 gene mutations were higher in COVID-19 (+) patients than COVID-19 (-) patients; p < 0.0001: OR (95%CI): 5.333 (2.359-12.16) and p = 0.0121 respectively. Yet, no significant relationship was found between mtDNA mutations and patients' age and biochemical parameters (p > 0.05). The results showed that the frequency of mtDNA mutations in COVID-19 patients is quite high and it is important to investigate the association of these mutations with other genetic mechanisms in larger patient populations.
Analysis of telomere length in patients with COVID-19 and investigation into its relationship with clinical- demographic data
(Cukurova Univ, Fac Medicine, 2023) Savrun, Atakan; Dirican, Ebubekir
Purpose: Novel coronavirus disease 2019 (COVID-19) is an infectious disease unknown before the 2019 outbreak in Wuhan. This study evaluated telomere length in COVID-19 (+) and (-) samples with clinical-demographic parameters.Materials and Methods: DNA was isolated from COVID-19 (+) (n=70) and (-) (n=70) patients. Telomere length was determined by real-time-PCR (RT-PCR). The 2- increment increment Ct method was used to analyze the telomere length of the samples.Results: There were significant differences in creatinine, LDH, ferritin, WBC, NEU and CRP in COVID-19 (+) patients compared to COVID-19 (-) patients. The NEU/LYM (or N/L) ratio was found higher in the patients with COVID-19 (+), than in COVID-19 (-). On the other hand, our COVID-19 (+) patients (mean +/- std:0.93 +/- 0.58) had significantly shorter telomere lengths than the COVID-19 (-) (mean +/- std:1.26 +/- 0.76). Moreover, COVID-19 (+) male patients (mean +/- std:1.06 +/- 0.50) had longer telomere length than female patients (mean +/- std:0.76 +/- 0.54). Telomere length was significantly shorter in patients with COVID-19 (+) with high blood urea nitrogen (BUN), high creatinine, high hematocrit, high NEU levels, normal platelets (PLT), and low WBC levels.Conclusion: Our findings suggest that telomere length and blood parameter levels influence the severity of COVID-19. Blood parameters differed in patients with COVID-19 (+) and COVID-19 (-). As a result, increasing the number of similar studies in the future can demonstrate the significance of our findings.
CYB mtDNA mutations and expression status of genes in the PI3K/AKT/mTOR signaling pathway in patients with schizophrenia
(Cukurova Univ, Fac Medicine, 2022) Dirican, Ebubekir; Uzuncakmak, Sevgi Karabulut; Ozcan, Halil
Purpose: This study aimed to screen for cytochrome b (CYB) mitochondrial DNA (mtDNA) mutations and analyze the mRNA expressions of genes in the PI3K/AKT/mTOR signaling pathway in patients with schizophrenia. Materials and Methods: In this study, whole blood was obtained from 44 schizophrenic patients and 41 healthy individuals for DNA (patients) and RNA (patients and control) isolation. Samples for CYB mtDNA mutations were amplified by PCR and identified by Sanger DNA sequencing. RT-PCR and 2- increment increment Ct method was used for mRNA expression of PIK3CA, AKT1, and mTOR genes. Results: In schizophrenia patients, m.15326 A > G (43/44), m.15452 C > A (5/44), m.15078 A > G (3/44), m.14872 C > T (3/44) and m.14798 T > C (3/44) was the most common CYB mtDNA mutations. In silico analysis showed that some of the mutations were associated with the harmful, disease-causing or benign character. The mRNA expression of the PIK3CA, AKT1, and mTOR genes in schizophrenia patients was significantly higher than in healthy individuals. There was a significant moderate positive correlation between the PIK3CA and AKT1 genes. In addition, by ROC analysis, PIK3CA, AKT1 and mTOR genes were found to have good diagnostic power in the patient group. ROC analyzes showed that PIK3CA in particular has significant diagnostic value for schizophrenia patients with 80% sensitivity and 63.4% specificity. Conclusion: Both of CYB mtDNA mutations frequency and PIK3CA, AKT1 and mTOR mRNA expression were higher in schizophrenic patients compared to healthy individuals. We believe that studying these mechanisms in
Determination of anti-cancer effects of Nigella sativa seed oil on MCF7 breast and AGS gastric cancer cells
(Springer, 2024) Cinar, Irfan; Gidik, Betul; Dirican, Ebubekir
Background This study aimed to investigate the cytotoxic, apoptotic, invasion, metastasis, and heat shock proteins (HSPs) effects of N. sativa oil on breast and gastric cancer cells. Methods We assessed the cytotoxic and apoptotic effects of various concentrations of N. sativa oil (10-50-100-200 mu g/mL) on MCF7 breast cancer and AGS, an adenocarcinoma of the gastric cell line, at 24, 48 and 72 h using the MTT test. Additionally, the expression of the Caspase-3, BCL2/Bax, MMP2-9 and HSP60-70 gene was examined using RT-PCR in cell lines treating with N. sativa. Results The MTT experiments demonstrate that N. sativa has a time and dose-dependent inhibitory effect on the proliferation of MCF7 and AGS cancer cells. The vitality rates of MCF7 and AGS cells treated with N. sativa were 77.04-67.50% at 24 h, 65.28-39.14% at 48 h, and 48.95-32.31% at 72 h. The doses of 100 and 200 mu g/mL were shown to be the most effective on both cancer cells. RT-PCR analysis revealed that N. sativa oil extract increased caspase-3 levels in both cell lines at higher concentrations and suppressed BCL2/Bax levels. Exposure of MCF7 and AGS cell lines to N. sativa caused a significant decrease in the expression of MMP2-9 and HSP60-70 genes over time, particularly at a dosage of 200 mu g/mL compared to the control group (p < 0.05). Conclusions Our findings indicate that N. sativa oil has a dose-dependent effect on cytotoxicity and the expression of apoptotic, heat shock proteins, and matrix metalloproteinases genes in breast and gastric cancer.
Effect of testicular morphology on embryo development to the blastocyst stage after round spermatid injection
(E-Century Publishing Corp, 2021) Ozmerdiven, Gokhun; Guler, Yavuz; Sahin, Eren; Tatar, Zeynep; Erbin, Akif; Dirican, Ebubekir; Haliloglu, Ahmet Hakan
If spermatozoa cannot be found after testiculer sperm extraction (TESE) in patients followed up due to nonobstructive azospermia (NOA) and the patients do not want donor spermatozoa, performance of round spermatid injection (ROSI) with the current technology seems to be the last resort. This retrospective study was conducted to evaluate the effect of testicular morphology on the development of embryos to the blastocyst stage obtained from ROSI. Between September 2019 and March 2020, after TESE and biopsy 29 patients who had only spermatid were taken to study. Tubular appearance, basal membrane appearance, Johnson score, peritubular fibrosis, interstitial fibrosis, and Leydig cell proliferation were pathologically examined. Following egg collection, ROSI was applied to the oocytes using the piezoelectric method. The embryos were monitored until the blastocyst stage. The mean age of the 29 patients was 36.3+5.01 years. Also, 7 patients had not previously undergone TESE, 20 had previously undergone once, and 2 had previously undergone twice. It was observed that having a history of TESE and a high Johnson score increased the likelihood of the embryo remaining in the blastocyst stage (P=0.021 and 0.014, respectively). However, other parameters do not affect the likelihood of blastocyst formation (P>0.05). Low TESE history and high Johnson score were associated with embryo development to the blastocyst stage. If spermatozoa are not found in patients with nonobstructive azoospermia, ROSI performed during initial TESE increases the likelihood of blastocyst formation.
Evaluating STC-1 and STC-2 mRNA expressions in Schizophrenia patients with increased oxidative stress
(Elsevier Masson s.r.l., 2023) Karabulut Uzunçakmak, Sevgi; Özcan, Halil; Dirican, Ebubekir; Tavacı Özçelik, Ayşegül
Introduction: Schizophrenia is a chronic psychiatric disorder characterized by cognitive impairment. Oxidative stress is associated with disease progression in patients with schizophrenia. Stanniocalcin (STC)-1 and STC-2 are two proteins commonly expressed in mammals belonging to the stanniocalcin family and they are involved in oxidative stress. The aim of this study is to investigate STC-1 and STC-2 mRNA expressions in schizophrenia patients with altered oxidative stress parameters. Methods: For this purpose, 70 patients with schizophrenia and 40 healthy individuals were recruited for the study. Peripheral blood samples were obtained from all participants. Glutathione (GSH) activities, superoxide dismutase (SOD) activities, and malondialdehyde (MDA) levels were measured. STC-1 and STC-2 expressions were assessed by real-time polymerase chain reactions. Results: SOD activity levels were lower in patients than in healthy individuals (p = 0.0309), while the patients’ MDA levels were higher (p = 0.039). STC-1 and STC-2 expressions were lower in patients than in healthy individuals (p = 0.1049 and p < 0.0001, respectively). Furthermore, these two genes had a positive correlation among the patients (r = 0.435, p = 0.0025). According to area under the curve (AUC) values, STC-2 (AUC = 0.8332, p < 0.0001) had better diagnostic power than STC-1 (AUC = 0.6167, p = 0.1037). Conclusions: The expression of stanniocalcins in schizophrenia was investigated here for the first time. Decreased STC-2 expression in patients with schizophrenia with increased oxidative stress parameters may guide the understanding of the pathophysiological mechanisms caused by oxidative stress, which may be increased in cognitive diseases such as schizophrenia, and it also has the potential to be a prognostic factor that can be used in the diagnosis of schizophrenia. © 2023 Elsevier Masson SAS
Evaluation Expression of the Caspase-3 and Caspase-9 Apoptotic Genes in Schizophrenia Patients
(Korean Coll Neuropsychopharmacology, 2023) Dirican, Ebubekir; Ozcan, Halil; Uzuncakmak, Sevgi Karabulut; Takim, Ugur
Objective: Apoptosis is programmed cell death that occurs by several pathways. Caspase-3 is induced by active caspase-9 via the intrinsic pathway. The aim of this research was to explore the expression of caspase-3 and caspase-9 in schizophrenia patients and healthy samples. Methods: RNA was isolated from the peripheral blood of 39 schizophrenia patients' and healthy samples. After cDNA synthesis, real time PCR (RT-PCR) was used to analyse caspase-3 and caspase-9 gene expression. The severity of psychopathological symptoms of schizophrenia was evaluated using the Positive and Negative Symptoms Scale for schizophrenia (PANSS) and Clinical Global Impressions (CGI). Results: The expression of caspase-3 and caspase-9 genes was higher in schizophrenia patients than in healthy samples (p = 0.012, p = 0.002, respectively). The increase in caspase-3 gene expression was significant with being male, smoking and with a duration of less than 6 years (p = 0.047, p = 0.049, p = 0.034, respectively). On the other hand, the increase in caspase-9 gene expression was significant in patients who is smoke, have children, and are under 33 years old (p = 0.040, p = 0.043, p = 0.045, respectively). A significant positive correlation was detected between the caspase-3 and caspase-9 gene expression (r = 0.3218, p = 0.049). Conclusion: Our findings indicate that caspase-3 and caspase-9 gene expression may activate cell death mechanisms by intrinsic apoptotic genes. Furthermore, caspase-3 and caspase-9 may play essential roles in different ways in schizophrenia. Hence there is a need to further study the apoptotic mechanism with expanded patient populations.
Evaluation of miR-130b-3p and miR-375 levels and telomere length with telomerase activity in prostate cancer
(Taylor & Francis Inc, 2024) Karadag, Abdullah; Dirican, Ebubekir; Ozmerdiven, Cagdas Gokhun; Ozen, Ata; Ayan, Semih; Kabadere, Selda
Prostate cancer (PC) is the most frequent cancer in males, as well as the second highest cause of cancer-related deaths in men. Differences in expression levels of miRNAs were linked with prostat cancer pathogenesis. qPCR was used to evaluate the expression of miR-130b-3p and miR-375 in Benign Prostate Hyperplasia (BPH (n = 20) and PC (n = 22, pre- and post-operative) patients plasma. Relative telomere lengths (RLTs) in genomic DNA isolated from plasma were measured with qPCR, and telomerase activity analyzed by the ELISA method. PSA levels of PC patients were greater than of BPH patients (p = 0.0473). miR-130b-3p and miR-375 levels were significantly lower in pre-operative specimens of PC patients according to BPH (p = 0,0362, p = 0.0168, respectively). Similarly, post-operative miR-375 levels were lower in PC patients than in BPH patients (p = 0.1866). BPH patients had shorter RTLs than PC patients in both pre- (p=0.0438) and post-operative (p=0.0297) specimens. Telomerase activity was higher in PC patients than BPH(p = 0.0129). Interestingly, telomerase activity was further increased after surgery (p = 0.0003). We aim to identify the levels of miR-130b-3p and miR-375 expression and their relationship with telomerase activity in PC patients. Our data suggest that miRNAs and telomere length (TL) with telomerase activity may play a role in regulating prostate tumorgenesis and may be used as biomarkers for PC diagnosis.
Expression level of the angiotensin 2 (ACE2) gene in patients with COVID-19
(Cukurova Univ, Fac Medicine, 2021) Dirican, Ebubekir; Aydin, Ismail Erkan; Savrun, Seyda
Purpose: Coronavirus disease-19 (COVID-19) is caused by the Severe Acute Respiratory Syndrome Coronavirus 2 (SARSCoV2). The angiotensin converting enzyme 2 (ACE2) can play a vital role in regulating the pathological changes of various diseases, including COVID-19. The aim of this study was to analyze the expression of the ACE2 gene in Turkish COVID-19 patients. Materials and Methods: Eighty COVID-19 PCR test positive patients and 80 PCR test negative healthy volunteers were included in the study. RNA isolation was carried out from all samples. The RT-PCR device was used to analyze the expression of the ACE2 gene. Results: A significant difference was found in the age, Lactate dehydrogenase (LDH), ferritin and C-reactive protein (CRP) levels of the COVID-19 patients compared to the healthy volunteers. Based on the gene expression results, it was found that ACE2 gene expression was significantly higher in COVID-19 patients compared to healthy volunteers. In patients with COVID-19, a significant difference was found between the expression of the ACE2 gene and the levels of Blood urea nitrogen (BUN), Hemoglobin (HGB), Hematocrit (HCT) and CRP. Conclusion: ACE2 gene expression was high in the COVID-19 patients and showed differences based on clinical data. Thus, the expression of ACE2 can have paradoxical effects that support the pathogenicity of SARSCoV2 but, conversely, limit viral infection. The availability of ACE2 receptors may increase the susceptibility and / or disease course of COVID-19.
HRM method for identification of TP53 exon 5 and 8 mutations in human prostate cancer patients
(Elsevier, 2022) Ozmerdiven, Cagdas Gokhun; Dirican, Ebubekir; Ayan, Semih; Tatar, Zeynep; Cakir, Sami; Guler, Yavuz; Karadag, Abdullah
Background: The purpose of the present study was to perform a high-resolution melting (HRM) analysis to discover mutations in gene exons 5-8 of tumor protein p53 (TP53), as well as the relationships of these mutations to clinical parameters in prostate cancer (PC).& nbsp;Methods: Genomic DNA was extracted from 50 formalin-fixed paraffin-embedded (FFPE) tissues with PC. Mutations in exons 5 and 8 of TP53 were analyzed using the HRM method. Sanger sequencing was used to describe mutations.& nbsp;Results: According to the HRM analysis results, 21 (42%) PC samples had different normalized and shifted melting curves from other samples. Mutations in TP53 exons 5 and8 were observed in 12 (24%) patients by the Sanger method. The detection sensitivity of the HRM method in exon 5 and exon 8 mutations was 66.7% and 50%, respectively. PSA levels of PC patients with TP53 mutation were found to be lower than that of patients with no mutation (p = 0.8270). However, we did not find any correlations between TP53 mutations and clinical parameters (p > 0.05).& nbsp;Conclusions: HRM analysis is a simple, rapid, and efficient mutation-scanning method for known/unknown mutations in TP53 exons 5and8, as well as an attractive method for detection of mutations and their analysis in FFPE tissues. Additional studies with larger patient populations are warranted to confirm the correlation between the TP53 mutations and PC risk.
Impact of gossypin on gene expression of HSP60 and HSP70 in different cancer cell lines
(Cukurova Univ, Fac Medicine, 2022) Dirican, Ebubekir; Cinar, Trfan
Purpose: The aim of this study is to evaluate the impact of gossypin on the expression level of heat shock proteins (HSPs) genes in different cancer cells. Materials and Methods: Cells were grown under standard culture conditions. Cancer cells were treated with different concentrations (5-100 mu g/ml) of gossypin and cisplatin (50 mu M) as positive control. Cell viability and effective dose range (5-100 mu g/ml) of gossypin were determined by MTT at 24, 48 and 72 hours. After RNA isolation and cDNA synthesis, HSP60 and HSP70 gene expression levels were analyzed using RT-PCR. For gene expression analysis, the 2-(Delta Delta ct) method was used. Results: According to the Mil results, 25-50-100 mu g/ml of gossypin doses were found effective on HSP60 and HSP70 gene expression levels in the cancer cell lines. Gossypin affected with dose-dependently the expression of HSP60 and HSP70 in the three cell lines. In the three cell lines, 50 mu g/ml and 100 mu g/ml of gossypin doses significantly reduced the expression of HSP60 and HSP70 compared to control group. Conclusion: Our results strongly supported the anticarcinogenic effect of gossypin at various doses in different cell lines. However, we believe that further in vivo research and human studies are needed. Our findings suggest that gossypin could be suitable candidate agent for further investigation to develop new strategies for the prevention and/or treatment of different cancer types.
Investigation of alterations in PIK3CA and OCT-4 gene expression in breast cancer
(Elsevier, 2020) Dirican, Ebubekir; Kankaya, Burak; Buyukasik, Suleyman; Alis, Halil; Velidedeoglu, Mehmet; Ilvan, Sennur; Ilvan, Ahmet
Breast cancer (BCa) is one of the most prevalent cancer amongst women and PIK3CA mutations have a crucial role in carcinogenesis. On the other hand, Octamer-Binding Transcription Factor 4 (OCT-4) is common biomarkers of cancer stem cells, which provide to the pathological processes of numerous carcinomas. In this study, we evaluated the association of OCT-4 gene expression levels with the PIK3CA mutation in BCa patients. The mRNA expression level of the OCT-4 gene was detected by qPCR in 44 BCa patients and 30 BCa patients control have positive PIK3CA mutations. Exon 9 and exon 20 of PIK3CA mutations were identified by sequencing. OCT-4 gene expression and PIK3CA mutations were also clinically evaluated in BCa patients. The frequency of PIK3CA mutation in 44 new cases were 32%. PIK3CA mutations were significantly associated with HER2-positive (p = 0.0271) status, Ki-67 status (p = 0.0434) and patient age (p = 0.0149). OCT-4 mRNA expression was significantly higher in the BCa tissue in comparison to normal breast tissue (p = 0.0305). A significant correlation was found between expression of OCT-4 and node status (p = 0.0190). OCT-4 mRNA expression was higher in the PIK3CA mutation tissue than the wild type tissue. However, no association was found between the expression of OCT-4 and the PIK3CA mutation or wild type samples (p = 0.7908). Our results provided that OCT-4 gene and PIK3CA mutations may have a role in development of BCa. OCT-4 mRNA expression level and PIK3CA mutations have a potential important for BCa diagnosis or treatment.
Investigation of the effect of gossypin on MMP-2 and MMP- 9 genes in prostate cancer cells
(Cukurova Univ, Fac Medicine, 2022) Dirican, Ebubekir; Cinar, Irfan
Purpose: The aim of this study is to explore the effects of gossypin on matrix metalloproteinases -2 (MMP-2) and MMP-9 genes in prostate cancer cells. Materials and Methods: PC3 cells were grown in culture and treated with three different concentrations of gossypin (25-50-100 mu g/ml) and cisplatin (50 mu M) as a positive control. First, RNA isolation was performed. Then, cDNA synthesis was performed and RT-PCR was used to evaluate mRNA expression levels of MMP-2 and MMP-9 genes. Results: Gossypin decreased MMP-2 and MMP-9 mRNA expression in prostate cancer cells in a concentrationdependent manner. Three concentrations (25-50-100 mu g/ml) of gossypin in PC3 cells reduced the mRNA expression of the MMP-2 gene. While the fold change value of MMP-2 gene expression was 0.3482 +/- 0.040 in the 100 mu g/ml gossypin group, it was 1.007 +/- 0.1425 in the control group. In addition, 50 mu g/ml and 100 mu g/ml concentrations of gossypin decreased the mRNA expression of the MMP-9 gene. The expression level of the MMP-9 gene in prostate cancer cells was 0.4740 +/- 0.038 in the 100 mu g/ml gossypin administered group, while it was 1.009 +/- 0.1687 in the control group. There was a positive correlation between the expressions of the MMP-2 and MMP-9 genes. Conclusion: According to the results obtained, it is seen that gossypin reduces the expression of MMP-2 and MMP9 genes in prostate cancer cells and the effects of gossypin on other genetic and epigenetic mechanisms in cancer need to be investigated to reveal the anti-cancer.
Investigation of the relationship between NFKB1 polymorphisms and telomere length and apoptosis in patients with type-2 diabetes
(Cukurova Univ, Fac Medicine, 2023) Dirican, Ebubekir; Kaya, Yasemin
Purpose: Type 2 diabetes mellitus (T2DM) is a heterogeneous, chronic, and metabolic disease that affects a significant proportion of the global population. This study aimed to evaluate the effect of NFKB1-94 ATTG ins/del polymorphisms on the expression of apoptosis genes and telomere length (TL) in patients with T2DM compared with healthy individuals.Materials and Methods: Sixty-nine T2DM patients and sixty healthy people were enrolled in the study. DNA and RNA were isolated from the blood samples. NFKB1 genotypes were identified by Sanger sequencing. For TL analyses and to investigate the expression of the caspase-3, caspase-9, bax, and bcl2 genes, RT-PCR was utilized.Results: There was a significant difference between the NFKB1-94 ins/del genotype patients and the control group (OR:0.4792 (0.2345-1.011)). However, the distribution of other genotype/alleles (ins/ins and del/del) showed no difference between T2DM and control groups. The allelic frequency of NFKB1-94 ins/del was 0.455/0.235 for the T2DM group and 0.435/0.165 for the control group. An increase in the mRNA expression of caspase-3, caspase-9 and Bax genes was observed in the T2DM group compared with the healthy group, while a decrease in the Bcl2 gene was found in the T2DM group. TL in T2DM patients was shorter than in healthy individuals.Conclusion: NFKB1-94 ins/del polymorphisms show significant differences in T2DM patients. We observed that apoptosis was activated and TL was shortened in patients with T2DM. However, no relationship between NFKB1 polymorphisms and apoptosis and TL could not be determined.
Meme Kanseri Hastalarında hTERT Gen Ekspresyonunun Klinikopatolojik Önemi
(2022) Dirican, Ebubekir; Kankaya, Burak; Tatar, Zeynep
Amaç: Bu çalışma, meme kanserinde insan telomeraz ters transkriptaz (hTERT) genin mRNA ekspresyonunun klinik önemini incelemeyi ve klinikopatolojik rolünü araştırmayı amaçlamıştır. hTERT geni hücre proliferasyonu, apoptoz ve hücre invazyonu gibi olaylarda rol almaktadır. Yüksek hTERT mRNA ekspresyonunun meme kanseri, mide, akciğer, glioblastoma, baş ve boyun gibi solid tümörlerde kötü prognoz ile ilişkili olduğu bildirilmektedir. Gereç ve Yöntemler: 59 meme kanser ve sağlıklı kan örneklerinin RNA izolasyonları gerçekleştirildi. hTERT geni mRNA ekspresyonu Gerçek Zamanlı Polimeraz Zincir Reaksiyonu (RT-PZR) yöntemi ile analiz edildi. mRNA ekspresyon sonuçları 2-??ctmetoduyla kanserli ve sağlıklı örneklerde değerlendirildi. Klinik veriler toplandı ve bu verilerin hTERT gen ekspresyonu ile ilişkisi istatiksel olarak değerlendirildi. Bulgular: Meme kanser hastalarında hTERT geni mRNA ekspresyonu sağlıklı örneklere kıyasla anlamlı derecede yüksekti (p=0,0211). Düşük hTERT geni ekspresyonu östrojen reseptör (ÖR) (+) ve insan epidermal büyüme faktör reseptörü 2 (HER 2) (+) olan hastalarda anlamlı derecede ilişkiliydi (sırasıyla p=0,0464; p=0,0080). hTERT geni ekspresyonu histolojik derece (grade) 3 hastalarda derece 2’ye kıyasla yüksek anlamlı bulundu (p=0,0389). Bununla birlikte hTERT ekspresyonun bazal benzeri histolojik tipte luminal A ve luminal B’ye kıyasla anlamlı derecede yüksek olduğu gösterildi (sırasıyla p=0,0160; p=0,0008). Sonuç: Çalışmamızın bulguları, meme kanser hastalarında hTERT gen ekspresyonunun sağlıklı örneklere göre anlamlı olarak daha yüksek olduğunu gösterdi. Anormal hTERT gen ekspresyonu kanserojenez veya teşhiste önemli bir rol oynayabilir.
Methoxy-activated indole-7-carbohydrazides; synthesis, antioxidant, and anticancer properties
(Wiley, 2023) Kandemir, Hakan; Izgi, Samet; Cinar, Irfan; Cebeci, Fatma; Dirican, Ebubekir; Saglam, Mehmet F.; Sengul, Ibrahim F.
Indole has been known as a key heterocyclic motif in the development of new structures for both chemical and biological properties. In this current study, a new range of indole-7-carbohydrazides has been successfully synthesized starting from the readily available 3-phenyl and 2,3-diphenyl 4,6-dimethoxyindoles. The structures of the novel compounds were confirmed by utilizing H-1 NMR, C-13 NMR, FT-IR, high-resolution mass spectrometry, and single crystal X- ray diffraction techniques. In addition, the indole-7-carbohydrazides showed promising antioxidant results in preliminary screens. Some of the new compounds generated from dimethoxy indoles were also screened for their anticancer activity against SH-SHY5Y (human neuroblastoma), AGS (human gastric adenocarcinoma), and MDA-MB-231 (human breast adenocarcinoma) cell lines. The results revealed that the compound 12 was the promising candidate, showing cytotoxic effects on both neuroblastoma, stomach, and breast cancer cells.
Preliminary investigation of gene expression levels of PAPP-A, STC-2, and HIF-1? in SARS-Cov-2 infected patients
(Springer, 2022) Uzuncakmak, Sevgi Karabulut; Naldan, Muhammet Emin; Dirican, Ebubekir; Kerget, Ferhan; Halici, Zekai
Background Coronavirus-19 is still considered a pandemic that influences the world. Other molecular alterations should be clearer besides the increasing cytokine storm and pro-inflammatory molecules. Hypoxic conditions that induce HIF-1 alpha lead to stimulate gene expression of STC-2 that targets PAPP-A expression. This study aimed to determine gene expression levels of PAPP-A, STC-2, and HIF-1 alpha in COVID-19 infection. We also aimed to reveal the relationship of these genes with laboratory and clinical data of COVID-19 patients. Materials and Results We extracted RNA from peripheral blood samples of COVID-19(+) and COVID-19(-) individuals. The real-time PCR method was used to measure mRNA expression of PAPP-A, STC-2, and HIF-1 alpha. Gene expression analysis was evaluated by the 2(-Delta Delta Ct) method. PAPP-A, STC-2, and HIF-1 alpha mRNA expressions of severe patients were higher than healthy individuals (p = 0.0451, p= 0.4466, p < 0.0001, respectively). Correlation analysis of gene expression patterns of severe patients demonstrated a positive correlation between PAPP-A and STC-2 (p <0.0001, r= 0.8638). Conclusion This is the first study that investigates the relation of PAPP-A, STC-2, and HIF-1 alpha gene expression in patients with COVID-19 infection. Besides the routine laboratory findings, PAPP-A, STC-2, and HIF-1 alpha mRNA expressions may be considered to patients' prognosis as a sign of increased cytokines and pro-inflammatory molecules.
Relation of ATPase6 Mutations and Telomere Length in Schizophrenia Patients
(Korean Coll Neuropsychopharmacology, 2023) Uzuncakmak, Sevgi Karabulut; Dirican, Ebubekir; Ozcan, Halil; Takim, Ugur
Objective: Schizophrenia is a serious mental disorder. Mutations in mitochondrial genes can change energy metabolism. Telomere is a tandem sequence at the end of chromosomes. Shorter telomere length has been shown in schizophrenia. The aim of this study was to determine the relationship between ATPase6 gene mutations and telomere length in schizo-phrenia patients.Methods: Blood samples of 34 patients and 34 healthy controls were used. In this study conventional PCR, Sanger sequencing technic and real-time PCR were utilized.Results: Five different mutations (A8860G, A8836, G8697A, C8676T, and A8701G) in the ATPase6 gene were identified in schizophrenia patients. The most seen mutation was A8860G (94%). Telomere length analysis indicated the relation of ATPase6 gene mutations and telomere length variations (p = 0.001). Patients carrying the A8860G mutation had shorter telomere lengths than patients carrying other mutations. Comparing telomere length between schizophrenia pa-tients and healthy controls revealed that the mean telomere length of schizophrenia patients was shorter than healthy controls (p = 0.006). The demographic analysis demonstrated a significant relationship between marital status and telo-mere length (p = 0.011). Besides that, the duration of the illness is another factor that impacts telomere length (p = 0.044). There is no significant relation between telomere length and other clinical and demographic characteristics including education status, age, gender, etc.Conclusion: In conclusion, telomere length and ATPase6 gene mutations have a significant relation. Studies with larger patient populations and investigation of other mitochondrial gene mutations will make the clearer link between telomere length and mitochondrial mutations.
Thiol disulfide homeostasis in primary dysmenorrhea
(Mre Press, 2022) Savrun, Atakan; Savrun, Seyda Tuba; Dirican, Ebubekir; Gokcen, Emre; Arici, Yeliz Kasko; Aygun, Ali; Korkmaz, Vakkas
Oxidative stress is defined as a result of the loss of balance between the production of free radical or reactive oxygen species and the antioxidant system. This study aimed to determine the level of thiol-disulfide homeostasis (TDH) in the serum of women with primary dysmenorrhea. The study group consisted of 42 subjects with primary dysmenorrhea, and the control group consisted of 30 volunteer women with demographic characteristics similar to the study group. Native thiol (SH), total thiol (TSH), disulphide (SS), and SH/SS parameters were measured for TDH of the subjects. The SH (p = 0.038) and SH/TSH (p = 0.046) levels were significantly higher while SS (p = 0.013), SS/SH (p = 0.042) and SS/TSH (p = 0.046) levels were lower in the study group than in the healthy control group. The SS cut-off value was determined as 17.85 in the study group (sensitivity = 61.9%, specificity = 43.3%). Therefore, the probability of dysmenorrhea may increase significantly when SS levels fall below this value. In subjects with dysmenorrhea, there was a decrease in SS levels and an increase in SH levels in order to protect the cells and tissues from the harmful effects of free radicals.

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