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    Chitinolytic Bacillus subtilis Ege-B-1.19 as a biocontrol agent against mycotoxigenic and phytopathogenic fungi
    (Walter De Gruyter Gmbh, 2019) Oztopuz, Ozlem; Sarigul, Nermin; Liaqat, Fakhra; Park, Ro-Dong; Eltem, Rengin
    Background: Biological control of pathogenic fungi is a possible alternate to the chemical control, which is harmful to humans and environment. Soil-borne Bacillus strains can be potential biocontrol agents and a source of lytic enzymes. Aim: This study aimed to examine biocontrol potential and lytic enzyme activities of a soil isolate Bacillus subtilis Ege-B-1.19. Materials and methods: Strain was identified by biochemical and 16S rRNA gene analysis and its biocontrol activity was investigated against Aspergillus niger EGE-K-213, Aspergillus foetidus EGE-K-211, Aspergillus ochraceus EGE-K-217, Fusarium solani KCTC6328, Rhizoctonia solani KACC40111 and Colletotrichum gloeosporioides KACC40689. Chitinase, chitosanase, N-acetyl-beta-hexosaminidase and protease activities of B. subtilis Ege-B-1.19 were also determined. Chitosanase was purified using Sephadex G-150 column and its molecular weight was determined by SDS-PAGE. Chitooligosaccharides production using chitosanase was carried out and analysed by TLC and HPLC. Results: Results depicted that B. subtilis Ege-B-1.19 has shown inhibitory effects against all the test fungi. Chitinase, chitosanase, N-acetyl-beta-hexosaminidase and protease activities were determined as 2.7 U mL(-1), 7.2 U 6.2 U mL(-1) and 12.2 U mL(-1), respectively. Molecular weight of purified chitosanase was 44 kDa. Chitosanase hydrolysed chitosan to glucosamine (GlcN), dimers (GlcN)(2) and trimers (GlcN)(3). Conclusion: Bacillus subtilis Ege-B-1.19 can be anticipated as useful biocontrol agent and its chitosanase can be utilized for enzymatic synthesis of chitooligosaccharides.
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    Effects of hyperbaric oxygen treatment on liver and kidney tissue in chronic arsenic toxicity
    (Undersea & Hyperbaric Medical Soc Inc, 2022) Eroglu, Huseyin Avni; Buyuk, Basak; Oztopuz, Ozlem; Makav, Mustafa; Aydeger, Cemre
    Arsenic (As) is a toxic substance that damages the human body through exposure to drinking water. This exposure damages many organs and tissues in the body, especially the liver and kidneys. Hyperbaric oxygen (HBO2) therapy is a treatment method that acts by reducing oxidative stress parameters in tissues with high-pressure oxygen. Based on this, our study aimed to investigate the effectiveness of HBO2 on liver and kidney tissues with chronic arsenic toxicity. In the study 24 male Wistar albino rats (220-300 g, two to three months old) were equally divided into four groups: Control; As; HBO2; and As+ HBO2. All animals were housed in individual cages. The toxicity model was created by adding arsenic to drinking water at a dose of 5 mg/kg/ day for 60 days. HBO2 was applied 2 ATA pressure for 90 minutes a day for five days. At the end of the study, liver and kidney tissues were taken and stored for analysis. In liver tissue, histopathological showed that arsenic reduced inflammatory cell infiltration, sinusoidal congestion, and hydropic degeneration, while HBO2 increased these measures. Similar results were found by TUNEL method. In kidney tissue, both histopathologic and TUNEL method examinations found similar results with the liver: The As group was more damaged than the As+ HBO2 group.
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    Effects of Melatonin on Cardiac Ion Channels in Rats with Metabolic Syndrome
    (Wiley, 2022) Ovali, Mehmet Akif; Oztopuz, Ozlem; Vardar, Selma Arzu
    [Anstract Not Available]
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    Expression levels of KCNQ1 and KCNQ3 genes in experimental epilepsy model
    (Cukurova Univ, Fac Medicine, 2020) Coskun, Ozlem; Oztopuz, Ozlem; Cokyaman, Turgay
    Purpose: Status epilepticus (SE) is a highly common neurological disease in children, with recurrent generalized convulsions for more than 30 minutes, and when not controlled, neuronal damage occurs in the brain. The aim of this study was to evaluate the changes in KCNQ1 and KCNQ3 gene expression levels in the acute period after SE. Materials and Methods: In rats, an experimental SE model was created with Li-Pc. Study; female Wistar albino [250-350 g, 21 (n=7)] rats were used in 3 groups as SE, control and sham groups. The total RNA obtained from brain tissues was converted to cDNA, and gene expression levels of KCNQ1 and KCNQ3 genes were assessed by quantitative real-time PCR (qRT-PCR) method. Results: Statistically in SE group; compared to the control and sham groups, a significant difference was observed in the gene expression levels of the KCNQ1 and KCNQ3 ion channels. The KCNQ1 and KCNQ3 gene expression levels of the experimental group was found higher than the other groups. Conclusion: Determining the changes in mRNA expression levels of genes encoding K+ ion channels will help to better understand the pathological mechanisms that occur during epilepsy. In the SE experimental model created, it is believed that an increase in mRNA expression of KCNQ1 and KCNQ3 will lead to drug therapy studies planned for the future.
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    Humic Acid Has Protective Effect on Gastric Ulcer by Alleviating Inflammation in Rats
    (Pleiades Publishing Inc, 2022) Sehitoglu, Muserref Hilal; Oztopuz, Ozlem; Karaboga, Ihsan; Ovali, Mehmet Akif; Uzun, Metehan
    The new agents are needed in treatment of gastric ulcer that have less side effects, adequate efficacy, and no drug interactions. In this study, we aimed to investigate the potential protective effects of humic acid on experimental gastric ulcer. Wistar Albino male rats (n = 48) were randomly divided into 8 groups as follow; Control (without any applications), Humic acid (50 mg/kg), ethanol group (1 mL/rat), and indomethacin group (25 mg/kg). In the treatment groups, both gastric ulcer model and humic acid 50 mg/kg were applied. In addition, famotidine the antiulcer drug was used as positive control. All medications were administered by oral gavage. Levels of ADAM10 and ADAMTS12 in gastric mucosa were determined by ELISA method. Hematoxylin-Eosin (H&E) staining, iNOS, and PCNA immunohistochemical staining were performed for histopathological investigations. Apoptosis was demonstrated by using the TUNEL method. In addition, the levels of inflammatory cytokines (TNF-alpha, IL-6, IL-10) and caspase-3 gene were determined by qRT-PCR. ADAM10 and ADAMTS12 levels significantly increased in the treatment groups compared to the ulcer groups (p < 0.05). The experimental groups showed mucosal erosion, bleeding, leukocyte infiltration and edema. Treatment with humic acid and famotidine was found to suppress iNOS activity, thereby decreasing proinflammatory activity and preventing damage to the gastric mucosa, while reducing the number of apoptotic cells. IL-6, IL-10, TNF-alpha and caspase-3 levels were significantly decreased in the treatment groups compared to damaged gastric mucosa. As a result, humic acid may be defined as a potential protective agent with its anti-inflammatory effect in gastric ulcer.
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    Hyperbaric oxygen mitigates KIM-1 and inflammatory cytokine levels in kidney transplantation
    (Undersea & Hyperbaric Medical Soc Inc, 2023) Buyuk, Basak; Oztopuz, Ozlem
    Background: The aim of this study was to investigate the effect of hyperbaric oxygen (HBO2) administration during cold ischemic time to organs removed from donors before kidney transplantation. A total of 24 rats were divided into three groups: Group 1 was the control group, Group 2 received 60 minutes of HBO 2 at 2.5 atmospheres absolute, and Group 3 received 120 minutes of 2.5 ATA HBO2. The renal artery was entered with a polyethylene catheter and perfused with a standard organ preservation solution. Falcon tubes containing organs obtained from rats in Groups 2 and 3 were placed in a box supported by ice blocks. The temperature was kept constant at 4 degrees C and the box was placed in a pressure tank with 2.5 ATA HBO2. HBO2 was applied for 60 and 120 minutes, respectively. Organ samples were harvested at the end of 24 hours for histopathological evaluation, immunohistochemical analysis of TNF-alpha and IL- 18, TUNEL analysis for apoptosis, and gene expression levels of kidney injury molecule-1 ( KIM-1) and caspase-3. In histopathological examinations, hematoxylin and eosin staining was performed and samples were evaluated for tubular necrosis and vacuolization criteria. Group 2 and Group 3 had significant decreases compared to Group one in this regard. Immunohistochemical staining was performed for TNF-alpha, IL-18, and apoptosis levels; significant decreases were found in Groups 2 and 3. There were significant decreases in Groups 2 and 3 for KIM-1 and caspase-3 gene expression levels compared to Group 1, as well. Thus, it was demonstrated that during the cold ischemic period before kidney transplantation, HBO2 administration to organs removed from donors can reduce apoptotic cell numbers, inflammatory cytokine release, and histopathological damage to the organs as well as decreasing the expression of the KIM-1 gene, which is an indicator of kidney damage.
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    Hyperbaric oxygen treatment ameliorates gentamicin-induced nephrotoxicity and expression of kidney injury molecule 1 in the rat model
    (Undersea & Hyperbaric Medical Soc Inc, 2019) Oztopuz, Ozlem; Turkon, Hakan; Sehitoglu, Muserref Hilal; Buyuk, Basak; Uzun, Metehan; Ovali, Mehmet Akif; Demir, Ufuk
    In recent years, hyperbaric oxygen (HBO2) therapy has been considered as an effective method for the treatment of gentamicin (GM)-induced renal toxicity. However, the findings related to the use of HBO2 for GM toxicity are limited and contradictory. The aim of this study is to investigate the protective role of HBO2 on GM-induced nephrotoxicity. For this purpose, Wistar albino rats (n=28) were randomly divided into four equal groups: C, HBO2, GM and GM+HBO2. GM (100 mg/kg, ip) and HBO2 were applied over seven days. On the eighth day blood and kidney tissue samples were harvested. The albumin, creatinine, and urea levels were determined from serum samples. Superoxide dismutase (SOD), glutathion peroxidase (GSH-Px) activities, malondialdehyde (MDA), total antioxidant status (TAS) and total oxidant status (TOS) values were analyzed spectrophotometrically. The relative expression level of TNF-alpha, IL-1 beta and Kim-1 gene were determined by qRT-PCR assays; histopathologic investigation was completed in kidney tissue samples. Serum urea, albumin and creatinine levels significantly increased in the GM group compared to the GM+HBO2 group. For antioxidant parameters the GM+HBO2 group was not statistically different from the C group but was significantly different compared with the GM group. TNF-alpha, IL-1 beta and Kim-1 gene expression levels in the GM group were statistically increased compared to the GM+HBO2 group (p=0.015, p=0.024, p=0.004) respectively. Severe tubular necrosis, epithelial desquamation and mild peritubular hemorrhage were observed in the GM-administrated group, while HBO2 exposure ameliorated these alterations. In conclusion, HBO2 exposure may be defined as a potential method for the prevention of GM-induced renal toxicity.
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    Isolation and Evaluation of New Antagonist Bacillus Strains for the Control of Pathogenic and Mycotoxigenic Fungi of Fig Orchards
    (Springer, 2018) Oztopuz, Ozlem; Pekin, Gulseren; Park, Ro Dong; Eltem, Rengin
    Bacillus is an antagonistic bacteria that shows high effectiveness against different phytopathogenic fungi and produces various lytic enzymes, such as chitosanase, chitinase, protease, and gluconase. The aim of this study is to determine Bacillus spp. for lytic enzyme production and to evaluate the antifungal effects of the selected strains for biocontrol of mycotoxigenic and phytopathogenic fungi. A total of 92 endospore-forming bacterial isolates from the 24 fig orchard soil samples were screened for chitosanase production, and six best chitosanolytic isolates were selected to determine chitinase, protease, and N-acetyl-beta-hexos-aminidase activity and molecularly identified. The antagonistic activities of six Bacillus strains against Aspergillus niger EGE-K-213, Aspergillus foetidus EGE-K-211, Aspergillus ochraceus EGE-K-217, and Fusarium solani KCTC 6328 were evaluated. Fungal spore germination inhibition and biomass inhibition activities were also measured against A. niger EGE-K-213. The results demonstrated that Bacillus mojavensis EGE-B-5.2i and Bacillus thuringiensis EGE-B-14.1i were more efficient antifungal agents against A. niger EGE-K-213. B. mojavensis EGE-B-5.2i has shown maximum inhibition of the biomass (30.4%), and B. thuringiensis EGE-B-14.1i has shown maximum inhibition of spore germination (33.1%) at 12 h. This is the first study reporting the potential of antagonist Bacillus strains as biocontrol agents against mycotoxigenic fungi of fig orchads.
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    Melatonin ameliorates cisplatin-induced neurodegeneration in medulla oblongata through the expressions of Aqp-1,-4, inflammation, and apoptosis pathway genes
    (Tubitak Scientific & Technological Research Council Turkey, 2022) Oztopuz, Ozlem
    In this study, the neuroprotective effects of melatonin (MEL) with changes in apoptosis, inflammation, and histopathological morphology were evaluated in the medulla oblongata of cisplatin (CIS) administered rats. Although the side effects of CIS are known in many tissues, its reaction on the medulla oblongata and the molecular association underlying this effect is unclear. Male wistar albino rats were separated into four groups (control, CIS, CIS+MEL, and MEL) (n = 24). CIS and CIS+MEL groups were given 4 mg/kg CIS at 4-day intervals (days 1, 5, 9, and 13) by the first day of the study. The MEL and CIS+MEL groups were given 10 mg/kg MEL daily for 13 days. At the end of the study, the medulla oblongata sections of the rats were harvested on the 14th day, and the changes in gene expressions were examined. Expression levels of inflammation markers (TNF-alpha and IL-6), apoptotic markers (Bax and Casp-3), and Aqp-1 and Aqp-4 were found to significantly increase with CIS administration. On microscopic examination, hemorrhage, edema, and perivascular edema were detected in the CIS applied group compared with controls. MEL treatment significantly reduced perivascular edema (p = 0.0152) and hemorrhage (p = 0.0087). Besides, there was a significant difference between the control and CIS groups regarding pyknosis and a significant increase in pyknotic neurons in the CIS treatment group (p < 0.001). This study indicates that CIS treatment significantly impaired medulla oblongata, and combined treatment with MEL ameliorates the injury in rats.
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    Melatonin ameliorates sodium valproate-induced hepatotoxicity in rats
    (Springer, 2020) Oztopuz, Ozlem; Turkon, Hakan; Buyuk, Basak; Coskun, Ozlem; Sehitoglu, Muserref Hilal; Ovali, Mehmet Akif; Uzun, Metehan
    Valproic acid (VPA) is a anticonvulsant and mood-stabilizing agent used to treat epilepsy in patients of all ages. However, it can cause hepatotoxicity with increased oxidative stress. Melatonin (MEL) is known as antioxidant and antiinflammatory agent. Therefore, the present study designed to investigate the probable protective role of melatonin against VPA-induced liver toxicity. For that purpose, 28 Wistar rats were randomly selected and divided into four groups, namely the Group C (vehicle), VPA (500 mg/kg/day VPA), MEL + VPA (10 mg/kg/day melatonin + 500 mg/kg/day VPA) and MEL (10 mg/kg/day melatonin). The agents were given by oral gavage for 14 days. Blood and liver tissue samples from all the rats were harvested on the 15th day of experiment. Biochemical analyses were conducted on the blood samples. The levels of malondialdehyde (MDA), superoxide dismutase (SOD), alpha glutathione S-transferases (alpha-GST), nuclear factor-kappa B (NF-kappa B), myeloperoxidase (MPO) and changes in gene expression were examined in the liver tissues. Also, liver histopathological analyses were conducted. VPA administration significantly increased the levels of alpha-GST, MDA, NF-kappa B and of IL-1 beta, TNF-alpha gene expression in the liver compared to Group C. Moreover, vacuolization, hydropic degeneration, inflammatory cell infiltration, and sinusoidal congestion were commonly detected in the VPA-treated group along with the highest apoptotic index (TUNEL staining) values. Melatonin administration was revealed to exhibit powerful protective properties at cellular, inflammatory and oxidative level activities against VPA-induced liver toxicity. Therefore, melatonin administration may be used as an adjuvant therapy against to VPA-induced liver toxicity.
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    Melatonin Improves Left Ventricular Mitochondrial Dynamics in Rats
    (Pleiades Publishing Inc, 2022) Uzun, Metehan; Oztopuz, Ozlem; Eroglu, Huseyin Avni; Doganlar, Oguzhan; Doganlar, Zeynep Banu; Ovali, Mehmet Akif; Demir, Ufuk
    There is increasing awareness that efficient and regular mitochondrial dynamics improvement cardiac function and affects the quality of life. Melatonin is a main pineal gland hormones and ameliorates mitochondrial dynamics in many cardiac disorders. For that purpose, we administrated melatonin to healthy rats all day long in order to investigate change in left ventricle mitochondrial dynamics both in the end of the nighttime and daytime. Twenty male Wistar rats (3-4 months age) were randomly assigned into Control (C; n = 10) and Melatonin groups (MEL; 10 mg/kg melatonin added drinking water, n = 10). On the 5th day of the study, 5 rats from the groups were randomly selected and euthanized at 08:00 AM and the remaining 5 rats were euthanized at 20:00 PM from each groups and samples of left ventricle (LV) tissue were harvested. Quantitative real-time PCR and western blot analysis demonstrated that melatonin acts preventive role on mitochondrial fusion and mitophagy through the DRP1/FIS1 and BNIP3/NIX axis, respectively. Additionally, melatonin administration significantly reduced P21 activation, induced cell cycle arrest, P27, finally regulated caspase-depended mitochondrial apoptosis signals in a time dependent manner. Our results suggest that melatonin may emerge as a therapeutic candidate to protect the bioenergetic dynamics of mitochondria in hearth.
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    Preoperative serum TNF-?, IL-6 and VEGF levels in patients with colorectal cancer
    (Elsevier Science Bv, 2017) Coskun, Ozlem; Oztopuz, Ozlem; Ozkan, Omer Faruk
    [Anstract Not Available]
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    The effect of Oleuropein on liver in experimental allergic rhinitis model
    (Bayrakol Medical Publisher, 2020) Oztopuz, Ozlem; Coskun, Ozlem; Buyuk, Basak
    Aim: The aim of this study is to investigate the supplemental effect of oleuropein which is the most commonly used corticosteroid in AR (Allergic rhinitis) treatment and to evaluate possible inflammatory-reducing properties in the liver. Material and Methods: The study was carried out on 42 male Wistar rats. The animals were randomly divided into 6 groups including G1 (C), G2 (AR), G3 (AR +SF), G4 (AR+MF), G5 (AR+OLE), G6 (AR+OLE+MF). Tumor necrosis factor-alpha (TNF-alpha) and Interleukin-1 beta (IL-1 beta) gene expression levels were assessed by RT-PCR analysis in liver tissues. In addition, liver tissues were evaluated histopathologically as a result of hematoxylin eosin (H&E) staining. Results: TNF-alpha and IL-1 beta gene expression levels in liver tissues were compared between groups and the results were statistically significant in all groups (p<0.000). Histopathological evaluation in H&E sections, sinusoidal dilatation, and hydropic degeneration were not observed in the liver tissue architecture of G1 (C). As a result of the examination of the sections belonging to G2 (AR), the location of hepatocyte cords and minimal destruction in portal areas were observed in organ architecture, and intensive sinusoidal dilatation was observed. Sinusoidal dilation was advanced in G3 (AR+SF). In addition, swelling and moderate hydropic degeneration were observed in hepatocytes, and focal lytic necrosis areas were not observed in G3 (AR+SF). All groups in terms of sinusoidal dilation and hydropic degeneration were significant differences statistically (p=0.000;0.001). Discussion: The present data indicated that oleuropein treatment suppresses inflammation in liver tissue caused by AR and it has no side effect in the experimental AR model. Oleuropein can be used as an auxiliary agent in the treatment of routine AR.
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    The Role of Melatonin in Oxidative Stress, DNA Damage, Apoptosis and Angiogenesis in Fetal Eye under Preeclampsia and Melatonin Deficiency Stress
    (Taylor & Francis Inc, 2019) Doganlar, Zeynep Banu; Guclu, Hande; Oztopuz, Ozlem; Turkon, Hakan; Dogan, Ayten; Uzun, Metehan; Doganlar, Oguzhan
    Aim: The aim of this study was to investigate the possible mechanisms of ocular damage induced by pinealectomy (PNX) and preeclampsia (PE), and to determine the cellular and molecular effects of melatonin treatment on oxidative stress, DNA damage, molecular chaperone responses, induction of apoptosis and angiogenesis in the fetal eye of both PNX and PNX+PE animals. Material and Methods: We analysed therapeutic potential of melatonin on fetal eye damage in PNX and PNX+PE animals using Malondialdehyde (MDA), Random Amplified Polymorphic DNA (RAPD), qRT-PCR and Western blot assays. Results: Our study presents three preliminary findings: (a) in fetal eye tissues, PNX and PNX+PE significantly induce oxidative damage to both DNA and protein contents, leading to a dramatic increase in caspase-dependent apoptotic signalling in both mitochondrial and death receptor pathways; (b) the same conditions trigger hypoxia biomarkers in addition to significant overexpression of HIF1-alpha, HIF1-beta, MMP9 and VEGF genes in the fetal eye; (c) finally, melatonin regulates not only the expression of genes encoding antioxidant enzymes and increase in DNA damage as well as lipid peroxidation but also limits programmed cell death processes in the fetal eye of PNX and PNX+PE animals . Furthermore, melatonin can relatively modulate genes in the HIF1 family, TNF-alpha and VEGF, thus acting as a direct anti-angiogenic molecule. In conclusion, both PNX and PNX+PE induce ocular damage at both cellular and molecular levels in fetal eye tissue of rats. Conclusion: Our results clearly indicate the potential of melatonin as a preventative therapeutic intervention for fetal ocular damage triggered by both PNX and PNX+PE.