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Öğe Avanafil as a Novel Therapeutic Agent Against LPS-Induced Acute Lung Injury via Increasing CGMP to Downregulate the TLR4-NF-?B-NLRP3 Inflammasome Signaling Pathway(Springer, 2022) Aydin, Pelin; Magden, Zeynep Berna Aksakalli; Uzuncakmak, Sevgi Karabulut; Halici, Hamza; Akgun, Nurullah; Mendil, Ali Sefa; Mokhtare, BehzadAim We demonstrate the effect of PDE5 inhibitors in cases of acute lung injury via the relationship between cGMP/NO and the TLR4-NF-kappa B-NLRP3 pathway. Materials and Methods This study was performed with 30 male Wistar albino rats. Lipopolysaccharide (LPS) was administered intratracheally to the rats and acute lung injury (ALI) was induced. Twelve hours after LPS administration, avanafil, prepared at suitable doses according to the body weights of the animals, was administered by oral gavage. Lung tissue samples of all groups were examined histopathologically and by immunochemical staining (IL-1 beta, iNOS, TLR4, and NF-kappa B). The iNOS, NLRP3, and IL-1B mRNA expression levels in the lung tissues were measured by RT-PCR. The left upper lobes of the rat lungs were dried at 70 degrees C for 48 h and lung water content was calculated. Result Statistically significant increases in iNOS, NLRP3, and IL-1 beta mRNA expressions were observed in the rats with ALI compared to the healthy controls (p < 0.0001). Those increased expressions were reduced at both doses of avanafil (p < 0.0001). This reduction was found to be greater at 20 mg/kg (p < 0.0001). IL-1 beta, iNOS, TLR4, and NF-kappa B immunopositivity was moderate/severe in the ALI group and mild in the group with ALI + avanafil at 20 mg/kg (p < 0.05). When the wet/dry lung ratios were calculated, a statistically significant increase was seen in the ALI group compared to the healthy rats (p < 0.05). That increase was decreased with both avanafil doses (p < 0.05). Conclusion We suggest that avanafil may prevent the progression of ALI and be effective in its treatment. We hope that this study will be supported by future clinical studies to yield a new indication for avanafil.Öğe Comparison of Serum Adropin Levels in Patients with Diabetes Mellitus, COVID-19, and COVID-19 with Diabetes Mellitus(Ataturk Univ, 2022) Aydin, Pelin; Uzuncakmak, Sevgi Karabulut; Tor, Ibrahim Hakki; Bilen, Arzu; Ozden, AyseObjective: In the present study, the relationship between a poor prognosis and adropin levels in diabetic patients with coronavirus disease 2019 was investigated by measuring serum adropin levels and levels of D-dimer, C-reactive protein, and ferritin, which are considered prognostic factors for coronavirus disease 2019. Materials and Methods: Hundred volunteer participants treated in the Erzurum Regional Training and Research Hospital were included in this study. Serum adropin levels were measured by enzyme-linked immunosorbent assay. The relationship between serum adropin level and C-reactive protein, ferritin, and D-dimer levels was analyzed by correlation analysis. Results: The participants' serum adropin levels differed between the groups (P=.0007). The control group had the highest adropin levels among groups. The lowest adropin levels were in the COVID + diabetes mellitus group. Adropin levels of diabetes mellitus, COVID, and diabetes mellitus + COVID groups were significantly decreased when compared to the control (P<.05). There was a significant negative correlation between adropin and C-reactive protein, D-dimer, and ferritin. Conclusion: Adropin can be used as an auxiliary biomarker, a prognostic factor in the early management of coronavirus disease 2019 patients with diabetes mellitus. We think that our study will guide future studies conducted in this field.Öğe 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, HalilPurpose: 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 inÖğe 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, UgurObjective: 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.Öğe Evaluation of IGFBP5 expression and plasma osteopontin level in COVID-19 patients(Elsevier Urban & Partner Sp Z O O, 2023) Uzuncakmak, Sevgi Karabulut; Aksakal, Alperen; Kerget, Ferhan; Aydin, Pelin; Halici, ZekaiPurpose: The aim of this study is to investigate insulin-like growth factor binding protein 5 (IGFBP5) expression in coronavirus disease 2019 (COVID-19) patients and its relationships with COVID-19 laboratory findings and plasma osteopontin (OPN) levels.Materials and methods: We enrolled 60 patients with COVID-19 and 30 healthy individuals in this study. mRNA expression of IGFBP5 was measured by RT-PCR. Plasma OPN levels were measured via the ELISA method.Results: Plasma OPN levels were higher and IGFBP5 expression levels were lower in COVID-19 patients than in the healthy individuals (p = 0.0057 and p = 0.0142, respectively). Critically ill patients had higher OPN and lower IGFBP5 than non-critically ill patients. Patients with affected lungs demonstrated increased OPN and decreased IGFBP5 (p = 0.00032 and p = 0.044, respectively). Receiver operating characteristic (ROC) analysis indicated that IGFBP5 expression and OPN levels can be used discriminate non-critically from critically ill patients (p = 0.049; p = 0.0016, respectively).Conclusion: This study demonstrated that patients with a poor prognosis had increased OPN and decreased IGFBP5. High values of OPN and low values of IGFBP5 may be considered as signs of disease severity. Tissue -specific IGFBP5 expression may contribute to understanding the role of IGFBP5 in the lungs in COVID-19 cases.Öğe LRIG1 levels in chronic rhinosinusitis with nasal polyps(Cukurova Univ, Fac Medicine, 2023) Uzuncakmak, Sevgi Karabulut; Sahin, Abdulkadir; Ozcelik, Aysegul Tavaci; Halici, ZekaiPurpose: Nasal polyps (NPs), usually occurring together with chronic rhinosinusitis (CRS), are benign masses of mucosal origin arising from inflammation. The transmembrane protein known as leucine-rich repeats and immunoglobulin-like domains 1 (Lrig1) is a member of the Lrig family. Lrig1 is frequently expressed in the respiratory tract and epithelial tissues and can inhibit several signaling pathways involved in cell proliferation. The aim of this study was to determine Lrig1 levels in NP tissues of patients with CRS. Material and Methods: This study included 36 patients with CRS and NPs and 15 patients who underwent rhinoplasty as the control group. The Lrig1 levels of all participants were measured by the ELISA method.Results: This study revealed that Lrig1 levels were significantly lower in NP tissues than in tissues of the control group. The mean level of Lrig1 of the NP tissues was 22.2 ng/ml, while the mean level of the control group was 28.5 ng/ml. According to the results of ROC analysis, Lrig1 levels have the power to distinguish polyp tissues from control tissues (AUC=0.794). Lrig1 levels were higher in tissues with scores of 4-8 than in tissues with scores of 16-20 based on the results of computed tomography scoring. According to endoscopic evaluations, Lrig1 levels of tissues with scores of 5-8 or 9-11 were relatively lower than those of tissues with scores of 2-4.Conclusion: Lrig1 levels were found to be decreased in NP tissues. Thus, Lrig1 may be used to confirm the presence of NPs. Lrig1 may also be helpful in NP grading. Increasing the Lrig1 levels in cases of NPs has the potential to become a targetable treatment modality.Öğe 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, ZekaiBackground 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.Öğe Relation of ATPase6 Mutations and Telomere Length in Schizophrenia Patients(Korean Coll Neuropsychopharmacology, 2023) Uzuncakmak, Sevgi Karabulut; Dirican, Ebubekir; Ozcan, Halil; Takim, UgurObjective: 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.Öğe Suberosin Alleviates Sepsis-Induced Lung Injury in A Rat Model of Cecal Ligation and Puncture(Taylor & Francis Inc, 2023) Uzuncakmak, Sevgi Karabulut; Halici, Zekai; Karakaya, Songul; Kutlu, Zerrin; Saglam, Yavuz Selim; Bolat, Ismail; Aydin, PelinBackground/aims Sepsis is one of the major problems encountered in intensive care units, causing organ damage and increasing mortality. Suberosin (SBR) is a type of coumarin with antioxidant and anti-inflammatory activities. The goal of this study is to explore the protective effects of SBR on the lungs in a rat model of sepsis. Methods Male Wistar rats were utilized in this study. A cecal ligation and puncture (CLP) model was applied to induce sepsis. Rats were separated into six groups with nine animals in each group, including healthy control, SBR, CLP, and CLP + SBR (5, 10, and 20 mg/kg) groups. Superoxide dismutase (SOD), glutathione (GSH) enzyme activities, and malondialdehyde (MDA) level were measured via enzyme-linked immunosorbent assay (ELISA). The messenger RNA (mRNA) expressions of tumor necrosis factor alpha (TNF-alpha) and interleukin 1 beta (IL-1 beta) were evaluated by real-time polymerase chain reaction (RT-PCR). Histopathological changes in the lungs were investigated with hematoxylin and eosin (H&E). Results MDA levels and GSH and SOD enzyme activities were negatively affected in the CLP group, but SBR treatment ameliorated these oxidative stress parameters in the SBR1-3 groups (p< 0.05). The mRNA expressions of TNF-alpha and IL-1 beta were increased in the CLP group, and SBR treatment decreased those expression levels in a dose-dependent manner (p < 0.05). Organ damage and necrosis were seen in the CLP group and were alleviated in the SBR3 group. Immunohistochemical (IHC) analysis of lung tissues demonstrated decreased TNF-alpha and IL-1 beta immunopositivity in the SBR1-3 groups (p< 0.05). Conclusions SBR ameliorated sepsis-related lung injury in a dose-dependent manner. This compound has significant potential as a future agent in the treatment of sepsis.
