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    3-Pyridinylboronic acid ameliorates rotenone-induced oxidative stress through Nrf2 target genes in zebrafish embryos
    (Springer, 2022) Üstündağ, Fümet Duygu; Ünal, İsmail; Üstündağ, Ünsal Veli; Cansız, Derya; Beler, Merih; Karagöz, Atakan; Kara Subaşat, Hülya; Alturfan, Ahmet Ata; Mega Tiber, Pınar; Emekli Alturfan, Ebru
    Parkinson's disease (PD) is one of the most common forms of neurodegenerative diseases and research on potential therapeutic agents for PD continues. Rotenone is a neurotoxin that can pass the blood-brain barrier and is used to generate PD models in experimental animals. Boron is a microelement necessary for neural activity in the brain. Antioxidant, non-cytotoxic, antigenotoxic, anti-carcinogenic effects of boric acid, the salt compound of boron has been reported before. Boronic acids have been approved for treatment by FDA and are included in drug discovery studies and pyridine boronic acids are a subclass of heterocyclic boronic acids used in drug design and discovery as substituted pyridines based on crystal engineering principles. The aim of our study was to determine the effect of 3-pyridinylboronic acid in rotenone-exposed zebrafish embryos, focusing on oxidant-antioxidant parameters and gene expression levels of nuclear factor erythroid 2-related factor 2 (Nrf2) target genes gclm, gclc, hmox1a, nqo1, and PD related genes, brain-derived neurotrophic factor, dj1, and tnfa alpha. Zebrafish embryos were exposed to Rotenone (10 mu g/l); Low Dose 3-Pyridinylboronic acid (100 mu M); High Dose 3-Pyridinylboronic acid (200 mu M); Rotenone + Low Dose-3-Pyridinylboronic acid (10 mu g/l + 100 mu M); Rotenone + High Dose-3-Pyridinylboronic acid (10 mu g/l + 200 mu M) in well plates for 96 h post-fertilization (hpf). Our study showed for the first time that 3-pyridinylboronic acid, as a novel sub-class of the heterocyclic boronic acid compound, improved locomotor activities, ameliorated oxidant-antioxidant status by decreasing LPO and NO levels, and normalized the expressions of bdnf, dj1, tnf alpha and Nrf2 target genes hmox1a and nqo1 in rotenone exposed zebrafish embryos. On the other hand, it caused the deterioration of the oxidant-antioxidant balance in the control group through increased lipid peroxidation, nitric oxide levels, and decreased antioxidant enzymes. We believe that these results should be interpreted in the context of the dose-toxicity and benefit-harm relationship of the effects of 3-pyridinylboronic.
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    3-Pyridinylboronic acid normalizes the effects of 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine exposure in zebrafish embryos
    (Taylor & Francis Ltd, 2022) Üstündağ, Fümet Duygu; Ünal, İsmail; Cansız, Derya; Üstündağ, Ünsal Veli; Kara Subaşat, Hülya; Alturfan, Ahmet Ata; Mega Tiber, Pınar; Emekli Alturfan, Ebru
    1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) is a neurotoxin that damages dopaminergic neurons. Zebrafish has been shown to be a suitable model organism to investigate the molecular pathways in the pathogenesis of Parkinson's disease and also for potential therapeutic agent research. Boron has been shown to play an important role in the neural activity of the brain. Boronic acids are used in combinatorial approaches in drug design and discovery. The effect of 3-pyridinylboronic acid which is an important sub-class of heterocyclic boronic acids has not been evaluated in case of MPTP exposure in zebrafish embryos. Accordingly, this study was designed to investigate the effects of 3-pyridinylboronic acid on MPTP exposed zebrafish embryos focusing on the molecular pathways related to neurodegeneration and apoptosis by RT-PCR. Zebrafish embryos were exposed to MPTP (800 mu M); MPTP + Low Dose 3-Pyridinylboronic acid (50 mu M) (MPTP + LB) and MPTP + High Dose 3-Pyridinylboronic acid (100 mu M) (MPTP + HB) in well plates for 72 hours post fertilization. Results of our study showed that MPTP induced a P53 dependent and Bax mediated apoptosis in zebrafish embryos and 3-pyridinylboronic acid restored the locomotor activity and gene expressions related to mitochondrial dysfunction and oxidative stress due to the deleterious effects of MPTP, in a dose-dependent manner.
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    520 nm and 660 nm light-emitting diodes modulates pancreatic development and beta cell functions in zebrafish embryos
    (2025) Üstündağ, Ünsal Veli; Ünal, İsmail; Cansız, Derya; Beler, Merih; Kanagaraj, Naveen Krishna; Kumar, Amrish Rajendra; Emekli Alturfan, Ebru
    Green and Red LEDs increase insulin production, but their comparative effects on pancreatic and beta cell development are unclear. Zebrafish embryos were divided into three groups: Control (n = 60), Green (G) (n = 60), and Red (R) (n = 60), then irradiated for three days (14 hours/day) with 0.5 W/cm2 G (λpeak = 520 nm, 180 mA) and R (λpeak = 660 nm, 210 mA). At the end of 72 h, pancreatic and beta cells, circadian rhythm, and oxidative stress gene were analyzed using RT-PCR. Malondialdehyde, nitric oxide, superoxide dismutase, and glutathione levels were also evaluated. In the Red group, pancreatic area increased by ~97.13% compared to the Control group and by approximately ~62.16% compared to the G group (both p < 0.0001), and no significant difference in beta cell area (p = 0.964). G group insulin expression increased 2.31-fold compared to R group (p < 0.0001). Red LED treatment increased MDA levels (p < 0.001), oxidative stress (fth1b, nqo1) (p < 0.0001), and per1b during the photophase (p < 0.0001) compared to G group. R LED treatment increases oxidative stress and disrupts circadian rhythm, leading to reduced insulin secretion. The positive effects of G LED treatment have potential for metabolic syndrome, diabetes, and pancreatic diseases.
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    Alcian blue staining for chondrocranium development in zebrafish
    (Humana Press Inc., 2024) Beler, Merih; Ünal, İsmail; Cansız, Derya; Emekli Alturfan, Ebru
    Craniofacial abnormalities are one of the most frequent birth malformations in humans, affecting around one in every thousand live births. The zebrafish (Danio rerio), a model organism that has seen increased usage in toxicological research in recent years, is ideal for assessing the effects of various chemicals on bone and cartilage structures. Chondrogenesis developed in zebrafish embryos by embryonic day 2, and supporting cartilage components are apparent at hatching (72 h post-fertilization). Individual cartilage may be observed using Alcian Blue staining as early as 2 days post-fertilization (dpf). The preferential binding of Alcian Blue causes the staining of zebrafish cartilage to acidic glycoproteins in an acidic solution (pH 2.2). In 72–120 hpf embryos, the cranial skeleton is easily visible after cartilage staining using Alcian Blue. Various cranial lengths and structures can be determined by measuring specific distances and angles to optimize the quantitative analysis of cranial malformations in zebrafish after exposure to various toxic agents. This chapter explains the Alcian Blue staining procedure to identify craniofacial cartilaginous structures in zebrafish embryos.
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    Altered tissue factor activity and disrupted oxidant-antioxidant status in saliva of patients with oral lichen planus
    (2024) Cansız, Derya; Beler, Merih; Ünal, İsmail; Eğilmezer, Gizem; Mızrak, Zülal; Emekli Alturfan, Ebru
    Objective: The aim of our study was to investigate salivary tissue factor (TF) activity and oxidant-antioxidant status, which is an important defense system, in saliva samples collected from patients with oral lichen planus (OLP), and to determine the relationship between the antioxidant system and TF activity in OLP. Materials and Methods: Saliva samples were collected from patients with OLP (n=20) and healthy subjects (n=13). TF activity, lipid peroxidation (LPO), nitric oxide (NO), glutathione (GSH) levels, glutathione S-transferase (GST), and superoxide dismutase (SOD) activities were measured. Results: NO levels and GST activities were increased, whereas GSH levels and SOD activities were decreased, when compared with healthy subjects in the saliva samples collected from patients with OLP. In addition, TF activity was increased in the OLP group compared with the control group Conclusion: The results revealed that the salivary oxidant-antioxidant balance was disrupted in parallel with the increase in TF activity in patients with OLP.
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    Amelioration of rotenone-induced alterations in energy/redox system, stress response and cytoskeleton proteins by octanoic acid in zebrafish: A proteomic study
    (Wiley, 2022) Sürmen, Mustafa Gani; Sürmen, Saime; Cansız, Derya; Ünal, İsmail; Üstündağ, Ünsal Veli; Alturfan, Ahmet Ata; Büyükkayhan, Derya; Emekli Alturfan, Ebru
    Rotenone is used to generate Parkinson's disease (PD)-like symptoms in experimental animals. Octanoic acid (C8), is the principal fatty acid of medium-chain triglycerides in ketogenic diets. Beneficial effects of ketogenic diets were shown in PD. We applied proteomic methods to reveal the effects of octanoic acid in rotenone toxicity in zebrafish to gain information on the use of ketogenic diets in PD. Zebrafish were exposed to 5 mu g/ml rotenone and octanoic acid (20 and 60 mg/ml) for 30 days. LC-MS/MS analysis was performed. Raw files were analyzed by Proteome Discoverer 2.4 software, peptide lists were searched against Danio rerio proteins. STRING database was used for protein annotations or interactions. 2317 unique proteins were quantified, 302 proteins were differentially expressed. Proteins involved in cell organization, biogenesis, transport, response to stimulus were most frequently expressed. Our study is first to report that the alterations in the expressions of proteins related to energy and redox system, stress response, and cytoskeleton proteins caused by rotenone exposure were normalized by octanoic acid treatment in zebrafish.
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    Assessment of dental caries and salivary nitric oxide levels in children with dyspepsia
    (BMC, 2019) Akşit Bıçak, Damla; Emekli Alturfan, Ebru; Üstündağ, Ünsal Veli; Akyüz, Serap
    BackgroundThe increase in nitric oxide (NO) levels in the oral cavity and saliva have been associated with various oral diseases; however, the gastro-salivary interaction of NO remains controversial. Thus, the aim of this study was to determine and compare salivary NO levels of dyspeptic and non-dyspeptic healthy children and to conduct an evaluation of its association with dental caries.MethodsSeventy children with dyspepsia (dyspeptic group) and 30 children without any gastrointestinal complaints (control group) were included in the study. Two biopsies from the gastric tissues were collected from dyspeptic children for histopathologic examination. Oral examination involved the assessment of dental caries, gingival index, plaque index, buffering capacity, salivary flow rate and pH. Salivary Streptococcus mutans (S. mutans) and Lactobacilli sp. counts were performed by commercial kits. For the comparison of the normal distribution between dyspeptic and control groups, Student t-test and for the comparison of the non-normal distribution, Kruskal-Wallis and Mann-Whitney-U tests were used. Chi-square test was used for comparison of qualitative data and the Pearson correlation test was used to evaluate the association between certain variables. Significance was assessed at p<0.05 level.ResultsHelicobacter pylori (H.pylori) were found in gastric biopsies of 84.2% (59/70) of the dyspeptic children. While the mean salivary NO values did not differ significantly between gastric H.pylori positive, negative and control groups, the salivary NO level of the dyspeptic group (213.751.68mol/dL) was found to be significantly higher than the control group (185.7 +/- 16.66mol/dL). No significant relationship was found between the mean salivary NO values, DMFT/dmft numbers and other oral parameters.Conclusions p id=Par4 The association of dental caries and salivary NO levels could not be considered specific in the current study. Although there were no statistically significant differences between salivary NO levels of gastric H.pylori positive, gastric H.pylori negative and control groups, greater salivary NO levels among dyspeptic children compared with the control group demonstrated that the concentration of NO in the saliva could be used as a biological marker in dyspepsia, which could lead to the improvement of more specified, uncomplicated and susceptible methods for analysis.
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    Assessment of developmental neurotoxicity using semi-automatic behavior analysis system for zebrafish
    (Humana Press Inc., 2024) Cansız, Derya; Ünal, İsmail; Beler, Merih; Alturfan, Ahmet Ata; Emekli Alturfan, Ebru
    Traditionally, morphological, conventional, and toxicological approaches have been used to demonstrate neurotoxicity; however, there has been a growing interest in animal behavioral methods for assessing neurotoxicity, both at the scientific and regulatory levels. Zebrafish (Danio rerio) is a small tropical freshwater fish currently recognized as a suitable model organism for investigating developmental neurotoxicity. There are many animal-tracking software programms used for behavioral analysis in biomedical research. Some of these software programms require a fee, which may exceed the laboratory budget and require detailed technical equipment. As a solution, freely available programs can be used. However, animal tracking may not be possible due to the glare from the aquatic environment of fish, and the small size of zebrafish embryos makes animal tracking difficult. In our laboratory, we developed a semi-automatic system to overcome these difficulties by using three different software available for free. This chapter explains the system for zebrafish embryos and adult zebrafish.
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    Bisphenol a and di(2-ethylhexyl) phthalate exert divergent effects on apoptosis and the Wnt/-catenin pathway in zebrafish embryos: A possible mechanism of endocrine disrupting chemical action
    (Sage Publications Inc, 2017) Üstündağ, Ünsal Veli; Ünal, İsmail; Ateş, Perihan Seda; Alturfan, Ahmet Ata; Yiğitbaşı, Türkan; Emekli Alturfan, Ebru
    Polyethylene terephthalate (PET) and polycarbonate (PC) are the most commonly used plastics in water bottles. Di(2-ethylhexyl) phthalate (DEHP) is used as a plasticizer in PET plastics, and bisphenol A (BPA) is used to produce PC. Both DEHP and BPA are known for their potential endocrine disrupting effects. The Wnt/-catenin signaling pathway has important roles in cell proliferation, cell specification and cell fate determination during embryonic development. Recent reports suggest a link between the Wnt/-catenin signaling pathway and apoptosis. The aim of this study was to investigate the relation between Wnt/-catenin signaling and apoptosis in the case of BPA and DEHP exposure in zebrafish embryos. Accordingly, in vivo cell death was assessed using acridine orange staining, and reverse transcription polymerase chain reaction was used to determine the expressions of wnt3a, gsk3 and ccnd1. Proliferative cell nuclear antigen, -catenin and Wnt3a expressions were determined immunohistochemically. Vitellogenin levels were determined using Enzyme Linked ImmunoSorbent Assay (ELISA). Increased vitellogenin levels, apoptosis, and wnt3a and gsk3 expressions were observed in BPA-exposed zebrafish embryos. Increased apoptosis in the BPA-exposed embryos may be due to the pro-apoptotic changes induced by Wnt3a, whereas DEHP might be suggested to have a minor effect as Wnt3a expression; vitellogenin levels and apoptosis did not increase significantly following exposure to DEHP.
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    Bisphenol a reveals its obesogenic effects through disrupting glucose tolerance, oxidant–antioxidant balance, and modulating inflammatory cytokines and fibroblast growth factor in zebrafish
    (SAGE Publications Ltd, 2022) Beler, Merih; Cansız, Derya; Ünal, İsmail; Üstündağ, Ünsal Veli; Dandin, Esra; Ak, Esin; Alturfan, A. Ata; Emekli Alturfan, Ebru
    Obesogens affect lipid metabolism, and genetic or epigenetic factors may also contribute to the progression of obesity. Endocrine-disrupting chemicals (EDCs) are the most striking among obesogens. Bisphenol A (BPA) is an estrogenic EDC used in food containers, adhesives, dye powders, and dental fillers. We aimed to elucidate molecular mechanisms of BPA’s obesogenic effects focusing on obesogenic pathways in the liver including fibroblast growth factor (FGF) and Dnmt3a which is its epigenetic regulator, oxidant-antioxidant status, and inflammatory cytokines. Zebrafish were divided into three groups as control, low-dose BPA (1 ?m BPA), and high-dose BPA groups (10 ?m BPA). At the end of 30 days, oral glucose tolerance test (OGTT) was performed, fasting blood glucose levels were measured, and hepatopancreas tissues were taken. Malondialdehyde (MDA) levels, superoxide dismutase (SOD), glutathione S-transferase (GST), and nitric oxide (NO) activities were examined in the hepatopancreas. Inflammatory cytokines, lepa, fgf21, and dnmt3a expressions were determined by RT-PCR. BPA exposure increased the body weights, il1ß, tnf?, il6, lepa, fgf21, and dnmt3a expressions, impaired glucose tolerance, and oxidant–antioxidant status in a dose-dependent manner. Hepatocyte degeneration, lipid vacuolization, and vasocongestion were observed in both BPA-exposed groups. Our study suggests impaired glucose tolerance, oxidant–antioxidant balance, increased inflammatory response, fgf21 expression, and dnmt3a expressions as the possible mechanisms for the BPA-induced obesity model in zebrafish.
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    Caprylic acid ameliorates rotenone induced inflammation and oxidative stress in the gut-brain axis in Zebrafish
    (Springer, 2021) Cansız, Derya; Ünal, İsmail; Üstündağ, Ünsal Veli; Alturfan, Ahmet Ata; Altınöz, Meriç A.; Elmacı, İlhan; Emekli Alturfan, Ebru
    Background Dysfunction of the gastrointestinal tract (GIT) is one of the most common non-motor symptom of Parkinson's Disease (PD). Pathological processes causing PD were suggested to initiate in the enteric nervous system (ENS) and proceed to the central nervous system (CNS). There are studies showing that low-carbohydrate ketogenic diets can improve motor symptoms of PD. Caprylic acid (C8) is the principal fatty acid component of the medium-chain triglycerides in the ketogenic diets. In this study, we aimed to evaluate the effects of caprylic acid, in neurotoxin exposed zebrafish focusing on the relationship between intestinal and brain oxidative stress and inflammation. Methods Adult zebrafish were exposed to rotenone (5 mu g/L) (R group) and caprylic acid (20 and 60 mg/mL) (L + HDCA and R + HDCA groups) for 30 days. At the end of 30 days locomotor activities were determined. Levels of lipid peroxidation (LPO), nitric oxide, glutathione and superoxide dismutase and glutathione S-transferase activities were determined by spectrophotometric methods and gene expressions of tnf alpha, il1, il6, il21, ifn gamma and bdnf were evaluated by RT-PCR in the brain and intestinal tissues of zebrafish. Results Caprylic acid ameliorated LPO, NO, SOD and the expressions of tnf alpha, il1, il6, il21, ifn gamma and bdnf in brain and intestines. Locomotor activities were only ameliorated in high dose R + HDCA group. Conclusions Caprylic acid ameliorated the neurotoxin-induced oxidative stress and inflammation both in the brain and intestines and enhanced locomotor activity in zebrafish.
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    Chard extract increased gastric sialic acid and ameliorated oxidative stress in valproic acid-administered rats
    (ScientificWebJournals (SWJ) Özkan Özden, 2023) Alev Tüzüner, Burçin; Tunalı, Sevim; Üstündağ, Ünsal Veli; İpekçi, Hazal; Emekli Alturfan, Ebru; Tunalı Akbay, Tuğba; Yanardağ, Refiye; Yarat, Ayşen
    Valproic acid, one of the most used drugs for epilepsy patients, has some known side effects. Chard extract has many pharmacological activities. The study aims to evaluate whether valproic acid might interfere with oxidative metabolism in gastric tissue and whether chard ameliorates these effects. The Sprague Dawley rats were divided into four groups (n=8); control, chard-given control, valproic acid, and chard-given valproic acid. The aqueous extracts of chard leaves were given 1 h before the administration of valproic acid for 7 days. On the 8th day, the animals were sacrificed under anesthesia and gastric tissues were homogenized. When compared to the control group, valproic acid significantly increased malondialdehyde and catalase activity, while superoxide dis- mutase activity decreased. Chard administration increased glutathione and, sialic acid levels and decreased malondialdehyde levels and superoxide dismutase activity in the valproic acid group. Based on these findings, since chard increased gastric sialic acid levels, we may suggest that chard may protect gastric mucosa may be through its antioxidant effects.
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    Determination of bisphenol a and phthalate levels in wastewater samples
    (Istanbul University, 2022) Akçay, Mansur; Ateş Kalkan, Perihan Seda; Üstündağ, Ünsal Veli; Ünal, İsmail; Cansız, Derya; Emekli Alturfan, Ebru; Alturfan, Ahmet Ata
    Objective: The use of endocrine-disrupting chemicals (EDCs) such as bisphenol A (BPA) in plastics manufacturing, agriculture, livestock, and paint manufacturing increas daily. The water treated in wastewater treatment plants is used in many areas such as irrigation of parks and gardens, and reinforcement of underground water resources. However, whether the treatment process eliminates EDCs in wastewater is not exactly known, and determining this as well as the amounts of these chemicals in treated water are important in terms of protecting the environment and human health. The aim of the study was to determine BPA and phthalate concentrations in the influent and effluent flow samples obtained from wastewater treatment plants. Materials and Methods: BPA and phthalate concentrations were measured in influent and effluent flow samples using the enzyme- linked immunosorbent assay (ELISA) method. BPA and phthalate measurements were performed as competitive measurements of BPA and total phthalates in samples using specific monoclonal antibodies. Results: BPA and phthalate levels were measured respectively as 7.69 ?g/L and 78.27 ?g/L in the influent water samples and 3.17 ?g/L and 25.56 ?g/L in the effluent water samples. The concentration of BPA and phthalates in the effluent samples decreased significantly compared to the influent water samples. Conclusion: This study is believed to shed light on the importance of monitoring BPA and phthalate concentrations in wastewater treatment plants and inspections for detecting other EDCs in wastewater.
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    Diethylhexyl phthalate exposure amplifies oxidant and inflammatory response in fetal hyperglycemia model predisposing insulin resistance in zebrafish embryos
    (2024) Kaplan, Gül; Beler, Merih; Ünal, İsmail; Karagöz, Atakan; Eğilmezer, Gizem; Üstündağ, Ünsal Veli; Cansız, Derya; Alturfan, Ahmet Ata; Emekli Alturfan, Ebru
    Exposure of zebrafish embryos to glucose is a suitable model for the fetal hyperglycemia seen in gestational diabetes. Diethylhexyl phthalate (DEHP), which is considered an endocrine-disrupting chemical, is one of the most common phthalate derivatives used in stretching plastic and is encountered in every area where plastic is used in daily life. In the present study, the effects of DEHP on pathways related to insulin resistance and obesity were examined in zebrafish embryos exposed to glucose as a fetal hyperglycemia model. Zebrafish embryos were exposed to DEHP, glucose, and glucose + DEHP for 72 h post-fertilization (hpf), and developmental parameters and locomotor activities were monitored. At 72 hpf ins, lepa, ppar?, atf4a, and il-6 expressions were determined by RT-PCR. Glucose, lipid peroxidation (LPO), nitric oxide (NO) levels, glutathione S-transferase (GST), superoxide dismutase (SOD), and acetylcholine esterase (AChE) activities were measured spectrophotometrically. Compared with the control group, glucose, LPO, GST activity, il6, and atf4a expressions increased in all exposure groups, while body length, locomotor, and SOD activities decreased. While AChE activity decreased in the DEHP and glucose groups, it increased in the glucose + DEHP group. Although glucose exposure increased ppar? and lepa expressions, DEHP significantly decreased the expressions of ppar? and lepa both in the DEHP and glucose + DEHP groups. Our findings showed that DEHP amplified oxidant and inflammatory responses in this fetal hyperglycemia model, predisposing insulin resistance in zebrafish embryos.
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    Differential effects of bisphenol a and di (2-Ethylhexyl) phthalate exposure on crestin and the expression of some genes related to apoptosis and inflammation in zebrafish embryos
    (Istanbul University Press, 2021) Ateş Kalkan, Perihan Seda; Ayık, Tuğçe; Üstündağ, Ünsal Veli; Emekli Alturfan, Ebru
    Objective: Endocrine disrupting chemicals (EDC) in plastics may disrupt proper endocrine system functioning. Zebrafish embryos are formed through external fertilization, and their rapid development, short life cycle, and transparency provide imaging advantages. In zebrafish embryos, neural crest development occurs similarly to other vertebrate embryos and crestin is found in the neural crest during embryogenesis. The aim of our study is to evaluate the effects of bisphenol A (BPA) and Di (2-ethylhexyl) phthalate (DEHP), which are the most widely used EDCs, on the expression of crestin, apoptosis, and inflammation-related parameters in zebrafish embryos. Materials and Methods: The embryos were exposed to either DEHP or BPA in well plates for 72 h post fertilization (hpf). Expressions of crestin were evaluated by in situ hybridization, while the expressions of bax, casp8, casp3a, ifng1, fas, and tp53 were evaluated by RT-PCR. Results: Expressions of bax and casp8 increased and casp3a, ifng1, and fas decreased in BPA and DEHP groups. tp53 expression increased in the BPA group but decreased in the DEHP group compared with the control group. In the DEHP group, casp3a, ifng1, fas, bax, casp8, and tp53 expressions decreased compared with the BPA group. No significant change was observed in the crestin expressions in the groups. When compared with the control group, an inverse relation between ifng1 expression and apoptosis, as evidenced by increased bax and casp8 expressions, was observed in the BPA and DEHP groups. Conclusion: Our study provided important data on the effects of EDCs on the relationship between inflammation and apoptosis.
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    Effects of benzoic acid synthesized from cinnamomum cassia by green chemistry on valproic acid-induced neurotoxicity in zebrafish embryos
    (2024) Cansız, Derya; Özokan, Gökhan; Bilginer, Abdulkerim; Işıkoğlu, Semanur; Mızrak, Zülal; Ünal, İsmail; Beler, Merih; Alturfan, A. Ata; Emekli Alturfan, Ebru
    Benzoic acid, the most basic aromatic carboxylic acid, is produced industrially and used in cosmetic, hygiene, and pharmaceutical items as a flavoring ingredient and/or preservative. The significance of sodium benzoate, a metabolite of cinnamon, used as a food preservative and FDA-approved medication to treat urea cycle abnormalities in humans, has been shown to raise the levels of neurotrophic factors. Valproic acid (VPA), a commonly used anti-epileptic and mood-stabilizing medication, causes behavioral and intellectual problems and is a commonly used agent to induce animal model for autism. Aim of this study is to determine the effects of benzoic acid synthesized from Cinnamomum Cassia by green chemistry method on gene expressions related to autism development in case of VPA toxicity. Zebrafish embryos were exposed to low and high doses of benzoic acid for 72 h post-fertilization. Locomotor activities were determined. Acetylcholinesterase (AchE), lipid peroxidation, nitric oxide (NO), sialic acid (SA), glutathione (GSH)-S-transferase, catalase (CAT), and superoxide dismutase (SOD) activities were determined spectrophotometrically. eif4b, adsl, and shank3a expressions were determined by RT-PCR as autism-related genes. Although high-dose benzoic acid inhibited locomotor activity, benzoic acid at both doses ameliorated VPA-induced disruption in oxidant-antioxidant balance and inflammation in zebrafish embryos and was effective in improving the impaired expression of autism-related genes.
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    Effects of exercise and calorie restriction on brain and testis in natural aging model
    (2021) Hakgüder, Umay; Üstündağ, Ünsal Veli; Tüzüner, Burçin Alev; Genç, Nevin; İpekçi, Hazal; Özçelik, Reyhan; Tunalı Akbay, Tuğba; Şener, Göksel; Yarat, Ayşen; Emekli Alturfan, Ebru
    The aim of our study was to examine the effects of exercise and calorie restriction on various tissue damage and antioxidant parameters in the brain and testis of rats in a natural aging model. For this purpose, male Sprague-Dawley rats were the control group (C), the elderly (A), the elderly with calorie restriction (ACR), the elderly who were exercised (AE) and the elderly who were exercised with calorie restriction (ACRE), they were divided into 5 groups. The control group was composed of three-month-old animals. The other group consisted of 15-month-old rats. Exercise and calorie restriction were applied for 6 weeks. At the end of the experiment, lipid peroxidation (LPO), nitric oxide (NO), glutathione (GSH) levels and superoxide dismutase (SOD), catalase (CAT), glutathione-S-transferase (GST) and tissue factor (TF) were determined in brain and testicular tissues homogenates. As a result of the study, the A group’s brain and testis LPO, NO levels and TF activity increased, GSH levels and SOD, CAT and GST activities decreased, when compared to the C group. As a result of our study, an increase in oxidant damage was observed with TF activity in the brain and testis in the natural aging model, and positive effects of exercise and calorie restriction on the antioxidant levels in the brain were determined, especially in aging.
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    Effects of low-dose ionizing radiation on the molecular pathways linking neurogenesis and autism spectrum disorders in zebrafish embryos
    (2024) Kollayan, Burcu Yeliz; Cansız, Derya; Beler, Merih; Ünal, İsmail; Emekli Alturfan, Ebru; Erçalık Yalçınkaya, Şebnem
    Prenatal exposure to environmental factors may play an important role in the aetiopathogenesis of autism spectrum disorder (ASD). We aim to investigate the potential effects of low-dose x-rays from dental diagnostic x-rays on neurodevelopment and molecular mechanisms associated with ASD in developing zebrafish embryos. Zebrafish embryos were divided into four groups and exposed using a dental x-ray unit: control, 0.08, 0.15 and 0.30 seconds, which are exemplary exposure settings for periapical imaging. These exposure times were measured as 7.17, 23.17 and 63.83 mSv using optical stimulated luminescence dosimeters. At the end of 72 hours post-fertilization, locomotor activity, oxidant-antioxidant status, and acetylcholine esterase (AChE) activity were analyzed. Expression of genes related to apoptosis (bax, bcl2a, p53), neurogenesis (α1-tubulin, syn2a, neurog1, elavl3) and ASD (eif4eb, adsl2a, shank3) was determined by RT-PCR. Even at reduced doses, developmental toxicity was observed in three groups as evidenced by pericardial edema, yolk sac edema and scoliosis. Deleterious effects of dental x-rays on neurogenesis through impaired locomotor activity, oxidative stress, apoptosis and alterations in genes associated with neurogenesis and ASD progression were more pronounced in the 0.30s exposure group. Based on these results we suggest that the associations between ASD and low-dose ionizing radiation need a closer look.
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    Endotoxin of porphyromonas gingivalis amplifies the inflammatory response in hyperglycemia-induced zebrafish through a mechanism involving chitinase-like protein YKL-40 analogs
    (Springer, 2023) Gündüz, Gizem; Beler, Merih; Ünal, İsmail; Cansız, Derya; Emekli Alturfan, Ebru; Köse, Kemal Naci
    Porphyromonas gingivalis (P. gingivalis), a key pathogen in periodontal diseases, is also associated with hyperglycemia-associated systemic diseases, including diabetes mellitus (DM). Gingipains are the most important endotoxins of P. gingivalis, and in vivo studies using gingipains are scarce. Zebrafish (Danio rerio) is a vertebrate with high physiological and genetic homology with humans that has multiple co-orthologs for human genes, including inflammation-related proteins. The aim of our study was to determine the effects of gingipain in a hyperglycemia-induced zebrafish model by evaluating inflammation, oxidant-antioxidant status, and the cholinergic system. Adult zebrafish were grouped into the control group (C), hyperglycemia-induced group subjected to 15 days of overfeeding (OF), gingipain-injected group (GP), and gingipain-injected hyperglycemic group (OF + GP). At the end of 15 days, an oral glucose tolerance test (OGTT) was performed, and fasting blood glucose (FBG) levels were measured. Lipid peroxidation (LPO), nitric oxide (NO), glutathione (GSH), glutathione S-transferase, catalase, acetylcholinesterase (AChE), alkaline phosphatase (ALP), and sialic acid (SA) levels were determined spectrophotometrically in the hepatopancreas. The expression levels of tnf-alpha, il-1 beta, ins, crp, and the acute phase protein YKL-40 analogs chia.5 and chia.6 were evaluated by RT-PCR. After two weeks of overfeeding, significantly increased weight gain, FBG, and OGTT confirmed that the zebrafish were hyperglycemic. Increased oxidative stress, inflammation, and AChE and ALP activities were observed in both the overfeeding and GP groups. Amplification of inflammation and oxidative stress was evident in the OF + GP group through increased expression of crp, il-1 beta, chia.5, and chia.6 and increased LPO and NO levels. Our results support the role of gingipains in the increased inflammatory response in hyperglycemia-associated diseases.
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    Öğe
    Evaluation of nitric oxide levels in chronic periodontitis patients treated with initial periodontal therapy and probiotic food supplements: a double blind, randomized controlled clinical trial
    (Taylor & Francis Ltd, 2019) İnce Kuka, Gizem; Gürsoy, Hare; Emekli Alturfan, Ebru; Üstündağ, Ünsal Veli; Kuru, Bahar
    The aim of the present study was to analyse the nitric oxide (NO) levels in gingival crevicular fluid (GCF) of patients with chronic periodontitis (CP) treated with probiotic-containing food supplements as an adjunct to initial periodontal therapy (IPT). The present study was a randomized, double blind clinical trial conducted in Yeditepe University Dental Hospital, Istanbul, Turkey. Thirty-six CP patients, with >= 2 nonmolar teeth in each quadrant with probing depth (PD) of 5-7 mm at proximal sites and radiologically detectable horizontal alveolar bone loss were randomized into two treatment groups. The test group received IPT + probiotic-containing food supplements, whereas the control group received IPT + placebo. GCF sampling was performed at the baseline and 3 and 6 months after treatment. The biochemical evaluation of NO in GCF was performed using the Griess colourimetric method. Statistical analysis was performed by using statistical software. Significant reductions in GCF volume and GCF NO levels were detected in each group after the treatments (p < 0.05). Intergroup comparison of the NO levels revealed statistically significant differences in favour of the test group at both 3 and 6-month evaluation periods (p < 0.05). Adjunctive usage of probiotic comprising food supplements to IPT yielded significant reduction of the NO levels compared to the control for up to 6 months of follow-up. The present study is registered at Thai Clinical Trials Registry (TCTR identification number: TCTR20171114003).