Determination of bisphenol a and phthalate levels in wastewater samples

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 μg/L and 78.27 μg/L in the influent water samples and 3.17 μg/L and 25.56 μg/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

The effect of different storage conditions on the migration of chemicals from polyethylene terephthalate and polycarbonate bottles to water

Objective: Polyethylene terephthalate (PET) and polycarbonate (PC) bottles have been used widely in the last years for the consumption of water and the increased use of these chemicals has raised many concerns regarding their adverse effects on health. Phthalates and bisphenol A (BPA) are the main endocrine disrupting chemicals (EDCs) that can migrate from these plastics into potable water. Materials and Methods: The concentrations of phthalate and BPA were measured in water samples that were stored in PET and PC bottles at different storage conditions. The method of ELISA was used for the determination of phthalate and BPA levels. A standard curve is obtained from the standards prepared at known concentrations of phthalate, BPA, according to their absorbance at 450 nm. The BPA levels of the samples were obtained through the calculation of the absorbance values acquired using the standard curve. Results: Different storage and heating processes applied on the samples significantly increased the levels of BPA and phthalate. One year of storage led to a statistically significant increase in phthalate levels when compared to the control group. Both BPA and phthalate levels detected in the water samples were higher than the control group depending on the storage conditions including exposure to high temperatures, sunlight and outdoor conditions. Conclusion: Our results indicate the necessity to establish the environmental conditions that must be ensured during the production, transportation and storage processes of the bottles, on a legal basis with legal regulations

Gingipain injection affects intestinal oxidant-antioxidant status and alkaline phosphatase in overfed zebrafish

Objective: Porphyromonas gingivalis (P. gingivalis), a major periodontopathogen, is associated with overfeeding disorders, including metabolic syndrome. Gingipains are one of the most powerful endotoxins of P. gingivalis. Our aim was to reveal the effects of gingipain injections on the intestinal oxidant-antioxidant status and alkaline phosphatase (ALP) activity in overfed zebrafish. Materials and Methods: Four groups of healthy adult zebrafish were placed in random tanks as C: Control (n=15); GP: Gingipain (n=15); OF: Overfeeding (n=15); and OF+GP: Overfeeding+Gingipain (n=15) groups. At the end of the experiment, levels of intestinal lipid peroxidation (LPO) and ALP, glutathione S-transferase (GST), and catalase (CAT) activities were evaluated. Results: Intestinal LPO was significantly lower in the GP and OF groups compared to C. Gingipain injection in OF (OF+GP) significantly elevated LPO when compared to C, GP, and OF groups. ALP activities decreased significantly in the GP, OF, and OF+GP compared to C. GST activities increased significantly in the GP when compared to C. Decreased GST activities were observed in the OF and OF+GP. This decrease was less in OF+GP. CAT activities significantly decreased in all groups when compared to C. Conclusion: Our findings demonstrate that gingipain injection alters the ALP activity and intestinal oxidant-antioxidant status in overfed zebrafish.Marmara Universit

Sodyum lauril sülfatın gelişmekte olan zebra balığı embriyolarında biyouyumluluğu

Objective: As an anionic surfactant, sodium lauryl sulphate (SLS), is used as an emulsifying agent in toothpastes as well as many different household cleaning products. Many toothpastes contain SLS and it is responsible for the formation of foam when brushing. However SLS may also irritate sensitive teeth and gums. Lipid peroxidation (LPO) is defined as an autoxidation process that is induced through the attack of free oxygen radicals leading to oxidative stress in the pathophysiology of various diseases. LPO causes the formation of highly reactive products including different aldehydes, ketones, and alkanes serving as biomarkers of LPO. In our study we aimed to expose zebrafish embryos to SLS and determine LPO in SLS exposed zebrafish embryos. Material and Method: Zebrafish embryos that were dividing normally and which had spherical shapes were chosen and they were exposed to SLS both in low and high concentrations in well plates for 72 hours. Rates of mortality and hatching were determined. The levels of malondialdehyde were evaluated using the Yagi's method as the end products of LPO in the form of thiobarbituric acid reactive substances. Results: Our findings showed LPO increased significantly in both low SLS (p<0.05) and high dose SLS (p<0.05) exposed zebrafish embryos when they were compared to the control group. Conclusion: It may be suggested that LPO is an early indicator of exposure to SLS during embryogenesis and further studies are required to confirm this finding.Amaç: Sodyum lauril sülfat (SLS), ev temizlik ürünleri ve diş macunlarında emülsifiye edici temizlik maddesi olarak kullanılan anyonik bir yüzey aktif maddedir. Birçok diş macunu fırçalarken köpük oluşumuna yol açan SLS içerir. Ancak SLS hassas diş ve diş etlerini tahriş edebilir. Lipid peroksidasyonu (LPO), çeşitli hastalıkların patofizyolojisinde oksidatif strese yol açan serbest radikallerin saldırısı ile başlatılan bir otooksidasyon sürecidir. LPO, LPO'nun biyobelirteçleri olarak görev yapan farklı aldehitler, ketonlar, alkanlar dahil olmak üzere reaktif ürünlerin oluşumuna neden olur. Çalışmamızın amacı, SLS'ye maruz kalan zebra balığı embriyolarında oluşan LPO'yu değerlendirmektir. Gereç ve Yöntem: Normal olarak bölünen küresel embriyolar, 72 saat boyunca plaka kuyucuklarında düşük ve yüksek dozda SLS'ye maruz bırakılmıştır. Mortalite ve kuluçkadan çıkma oranları belirlenmiştir. Tiyobarbitürik asit reaktif maddeler olarak LPO'nun son ürünü olan malondialdehit (MDA) düzeyini belirlemek için Yagi yöntemi kullanılmıştır. Bulgular: Bulgularımız, hem düşük SLS (p<0,05) hem de yüksek doz SLS’ye (p<0,05) maruz kalan zebra balığı embriyolarında kontrol grubu ile karşılaştırıldığında LPO'nun önemli ölçüde arttığını göstermiştir. Sonuç: LPO'nun embriyogenez sırasında SLS'ye maruz kalmanın erken bir göstergesi olduğu öne sürülebilir ve bu bulguyu doğrulamak için farklı çalışmaların yapılması gerektiği düşünülmektedir

Sodium-dependent glucose co-transporter-2 inhibitor empagliflozin exerts neuroprotective effects in rotenone-induced Parkinson's disease model in zebrafish; mechanism involving ketogenesis and autophagy

Sodium-dependent glucose co-transporter-2 (SGLT2) inhibitor empagliflozin (EMP), is the new class of oral hypoglycemic agent approved as a treatment for Type 2 diabetes. SGLT2 inhibitors may induce ketogenesis through inhibiting the renal reabsorption of glucose. In recent years, positive effects of ketogenic diets on neurodegenerative diseases such as Parkinson's disease (PD) have been reported by improving autophagy. We aimed to evaluate the effects of EMP treatment as a SGLT2 inhibitor that can mimic the effects of ketogenic diet, in rotenone induced PD model in zebrafish focusing on ketogenesis, autophagy, and molecular pathways related with PD progression including oxidative stress and inflammation. Adult zebrafish were exposed to rotenone and EMP for 30 days. Y-Maze task and locomotor analysis were performed. Neurotransmitter levels were determined by liquid chromatography tandem- mass spectrometry (LC-MS/MS). Lipid peroxidation (LPO), nitric oxide (No), alkaline phosphatase, superoxide dismutase, glutathione, glutathione S-transferase (GST), sialic acid, acetylcholinesterase, and the expressions of autophagy, ketogenesis and PD-related genes were determined. Immunohistochemical staining was performed for the microglial marker L-plastin (Lcp1) and tyrosine hydroxylase (Th). EMP treatment improved DOPAC/DA ratio, Y-Maze task, locomotor activity, expressions of Th and Lcp-1, autophagy and inflammation related (mTor, atg5, tnfα, sirt1, il6, tnfα); PD-related (lrrk2, park2, park7, pink1), and ketone metabolism-related genes (slc16a1b, pparag, and pparab), and oxidant-damage in brain in the rotenone group as evidenced by decreased LPO, No, and improved antioxidant molecules. Our results showed benefical effects of EMP as a SGLT2 inhibitor in neurotoxin-induced PD model in zebrafish. We believe our study, will shed light on the mechanism of the effects of SGLT2 inhibitors, ketogenesis and autopahgy in PD.Marmara Universit

Morphine attenuates neurotoxic effects of MPTP in zebrafish embryos by regulating oxidant/antioxidant balance and acetylcholinesterase activity

Parkinson's disease (PD) is one of the most common neurodegenerative diseases due to the loss of dopaminergic neurons in the midbrain in the substantia nigra. 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) is a neurotoxic agent causing disruptions in mitochondria of dopaminergic neurons leading to impaired oxidant-antioxidant balance. Both zebrafish and zebrafish embryos are sensitive to MPTP. In zebrafish embryos, MPTP decreases the dopaminergic cells in the diencephalon by damaging dopaminergic neurons. Morphine is an opioid pain killer and a strong analgesic that is used to treat chronic pain. Until today morphine has been shown to regulate the survival or death of neurons and both protective and destructive effects of morphine have been reported in the central nervous system. This study aimed to evaluate the effects of morphine in MPTP-exposed zebrafish embryos. Developmental parameters were monitored and documented daily during embryonic development. Locomotor activity of zebrafish embryos at 96 h postfertilization (hpf) was determined. Acetylcholinesterase (AChE) activity and oxidant-antioxidant parameters were analyzed by biochemical methods. RT-PCR was used to evaluate bdnf, dj1, lrrk and pink1 expressions. Morphine treatment improved mortality and hatching rates, locomotor activity, AChE, and antioxidant enzyme activities as well as the expressions of bdnf, dj1, lrrk and pink1 in a dose-dependent manner that were altered by MPTP. Increased lipid peroxidation supports the role of morphine to induce autophagy to prevent PD-related pathologies. Our study provided important data on the possible molecular mechanism of the therapeutic effects of morphine in PD

Amelioration of rotenone-induced alterations in energy/redox system, stress response and cytoskeleton proteins by octanoic acid in zebrafish: A proteomic study

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.Scientific Research Projects Coordination Unit of University of Health Science

The effect of acetic acid-induced pain in Parkinson's disease model in zebrafish

Parkinson's disease (PD) is the second most common neurodegenerative disease caused by the degeneration of dopaminergic neurons and the accumulation of Lewy bodies. Pain is one of the most common non-motor symptoms in PD, but the molecular mechanism of pain in PD is not fully understood, which prevents early diagnosis of PD. We aimed to determine the changes in opioidergic pathways when external pain is inflicted by inducing pain intraperitoneally in zebrafish, for which we generated a rotenone-induced PD model. After behavioural analyses in control(C), acetic acid (AA), rotenone (ROT), and rotenone+ acetic acid (ROT+AA) groups, catecholamine levels in brain tissue were determined by LC-MS/MS, expression of opioid peptides and their receptors by RT-PCR, expression of tyrosine hydroxylase by immunohistochemical method, and analyses of oxidant-antioxidant parameters by spectrophotometric methods. In the ROT group, distance travelled, average speed, and brain dopamine levels decreased, while LPO (lipid peroxidation) and NO (nitric oxide) increased as indicators of oxidative damage, and the SOD activity decreased. The mRNA expression of lrrk, pink1, and park7 genes associated with PD increased, while the mRNA expression of park2 decreased. This indicates that rotenone exposure is a suitable means to induce PD in zebrafish. The fact that body curvature was higher in the AA group than in the ROT and ROT+AA groups, as well as the decreased expression of penka, pdyn, and ion channels associated with the perception of peripheral pain in the ROT+AA group, suggest that mechanisms associated with pain are impaired in the rotenone-induced PD model in zebrafish.Istanbul University-Cerrahpas

Caprylic acid ameliorates rotenone induced inflammation and oxidative stress in the gut-brain axis in Zebrafish

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

Bisphenol a reveals its obesogenic effects through disrupting glucose tolerance, oxidant–antioxidant balance, and modulating inflammatory cytokines and fibroblast growth factor in zebrafish

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 μm BPA), and high-dose BPA groups (10 μm 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, tnfα, 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.Marmara Universit