Assessment of Protective Effects of DTPA, NAC, and Taurine on Possible Cytotoxicity Induced by Individual and Combined Zinc Oxide and Copper Oxide Nanoparticles in SH-SY5Y Cells.

The present study investigated the cytotoxic effects of ZnO, CuO, and mixed combinations of them on SH-SY5Y cells. For this purpose, the cells were exposed to various concentrations of these NPs alone for 24-96 h and as a mixture for 24 h. Variations in cell viability were noted. MTT results showed that ZnO and/or CuO NPs decreased cell survival by about 59% at 200 (ZnO, at 24 h) and 800 µg/ml (ZnO and/or CuO, at 72 and 96 h). When the NR assay was used, slight decreases were noted with ZnO NPs at 72 and 96 h. With CuO NPs alone and NPs in a mixture, only the highest concentrations caused 40 and 70% decreases in cell survival, respectively. Especially with NR assays, DTPA, NAC, or taurine provided marked protection. ROS levels were increased with the highest concentration of CuO NPs and with all concentrations of the mixture. The highest concentration of ZnO NPs and the lowest concentration of CuO NPs caused slight decreases in mitochondrial membrane potential levels. Additionally, increases were noted in caspase 3/7 levels with ZnO and CuO NPs alone or with a mixture of them. Intracellular calcium levels were decreased in this system. These findings demonstrated that ZnO and CuO NPs, either separately or in combination, had a modest cytotoxic effect on SH-SY5Y cells. Protection obtained with DTPA, NAC, or taurine against the cytotoxicity of these NPs and the ROS-inducing effect of CuO NPs and the NPs' mixture suggests that oxidative stress might be involved in the cytotoxicity mechanisms of these NPs.

The relationship between environmental air pollution, meteorological factors, and emergency service admissions for epileptic attacks in children.

OBJECTIVE: The aim of this study is to investigate the relationship between exposure to acute air pollution and meteorological factors on the frequency of epileptic attacks in children. METHODS: This retrospective study was carried using patient files from a children's hospital in Diyarbakir, one of the largest cities in Turkey. In the present study, the possible relationship between epileptic attacks seen in children over a 10-year period, two air polluting factors (PM10 and SO2), and the meteorological factors (air pressure, humidity, precipitation, wind speed) affecting them were investigated. The effects of different variables on the number of epilepsy patients admitted to the pediatric emergency department were also evaluated through four different models utilizing Poisson Regression Analysis. RESULTS: According to Model 2 and 3, the strongest relationship of the four Poisson Regression models, there was a significantly increased risk of pediatric emergency department admissions for seizures associated with a 10 µm/m3 increase in PM10 (IRR=1.020; 95% CI: 1.018-1.022); IRR= 1.071; 95% CI: 1.050-1.081; respectively) and 10 µm/m3 increase in SO2 (IRR=1.162; 95%CI: 1.151-1.173; IRR=1.092; 95% CI: 1.042-1.120; respectively). In Model 2, a 1 m/s increase in wind speed decrease the risk of daily of epileptic attack admitted to the emergency department and a 1 °C increase in temperature increased the risk of daily of epileptic attack admitted to the emergency department (IRR=0.840; 95% CI; 0.714-0.987; IRR=1.033; 95%CI: 1.007-1.059; respectively). In Model 3, 1% increase in humidity and 1 m/s increase in wind speed increased the number of daily epileptic attack admitted to the emergency department (IRR=1.008; 95%CI: 1.004-1.011; IRR=1.169; 95%CI: 1.056-1.294; respectively). The daily number of epilepsy patients was statistically significantly affected by the autumn (95%CI: 10.017-19.845) and winter (95%CI: -0.279 to 13.292) seasons. CONCLUSION: Meteorological factors and air pollutants affect the number of pediatric patients admitted to the pediatric emergency department with epilepsy attacks.

Investigation of the toxicity of a glyphosate-based herbicide in a human liver cell line: Assessing the involvement of Nrf2 pathway and protective effects of vitamin E and α-lipoic acid.

Glyphosate-based herbicides (GBHs) are the most widely used herbicides all over the world and has gained more attention in recent years because of health safety concerns. In this study, Roundup, one of the most popular glyphosate formulations, was used to evaluate cytotoxic, oxidative stress and apoptosis inducing effects of GBHs in a human hepatocellular cell line (HepG2). Roundup was shown to significantly increase cellular reactive oxygen species (ROS) levels, which lead to activation of the nuclear factor-erythroid-2-related factor 2 (Nrf2) antioxidant defense pathway including reduced levels of heme oxygenase 1 (HO-1). Furthermore, Roundup was found to induce apoptosis and further analysis confirmed involvement of a mitochondrial-dependent pathway verified by increased Bax/Bcl-2 ratios. Investigation of the protective effects of antioxidants vitamin E (Vit E) and α-lipoic acid (LA) against Roundup toxicity showed that both antioxidants significantly reduced the cytotoxicity, ROS formation, HO-1 downregulation, and apoptosis and that Vit E did so more efficiently than LA. In conclusion, our findings highlight the ROS producing and apoptosis inducing effects associated with GBHs, the activation of Nrf2 pathway as a defense mechanism and the protective effects of Vit E and LA against GBH toxicity.

Perfluorooctanoic acid affects mouse brain and liver tissue through oxidative stress.

The aim of this study was to investigate oxidative stress induced by perfluorooctanoic acid (PFOA) in the brain and liver tissues of Balb/c mice as well as protective effects of taurine and coenzyme Q10 (CoQ10) in both organs. For this purpose, animals were treated with PFOA (15 and 30 mg/kg) orally and their lipid peroxidation, total glutathione levels (GSH), and antioxidant enzyme activities measured and both tissues analysed for histopathological changes. Our results showed a dose-dependent decrease in body weight and increase in relative brain and liver weights, PFOA-induced lipid peroxidation and reduced glutathione peroxidase (GPx) activity in the brain tissue, and changes in GSH levels, GPx, superoxide dismutase (Cu-Zn SOD), and catalase (CAT) activities in the liver tissue. Pre-treatment with taurine or CoQ10 provided protection against PFOA-induced Cu-Zn SOD reduction in the liver tissue. Our findings evidence the depleting effect of PFOA on antioxidative systems and confirm that PFOA exerts its (neuro)toxicity through oxidative stress, but further research is needed to identify the exact toxicity mechanisms, especially in the brain.

Assessment of perfluoroalkyl substances levels in tap and bottled water samples from Turkey.

Perfluoroalkyl and polyfluoroalkyl substances (PFASs) draw considerable attention for their potential toxic effects in humans and environment. Drinking water is accepted as one of the major exposure pathways for PFASs. In this study, we measured concentrations of 10 perfluoroalkyl substances in 94 tap water samples collected in two different sampling periods (August 2017 and February 2018) from 33 provinces of Turkey, as well as in 26 different brands of plastic and glass-bottled water samples sold in supermarkets in Turkey. Perfluorohexanoic acid (PFHxA), perfluorobutane sulfonate (PFBS) and perfluoropentanoic acid (PFPeA) were the most frequently detected PFASs in the samples of tap waters. The maximum concentrations in tap waters were measured as 2.90, 2.37, 2.18, 2.04, and 1.93 ng/L, for PFHxA, perfluorooctanoic acid (PFOA), perfluorohexane sulfonate (PFHxS), perfluorooctane sulfonate (PFOS), and perfluorobutanoic acid (PFBA), respectively. The most abundant perfluorinated chemical in tap water samples was PFBA with 17%, followed by PFOS (13%), PFBS (12%), perfluoroheptanoic acid (PFHpA) (11%), PFHxA (11%), and PFOA (11%). The total PFASs concentration in tap water ranged from 0.08 to 11.27 ng/L. As regards bottled waters, the concentrations of PFASs were generally lower than those in tap water samples. These results revealed that tap water samples in Turkey might be considered generally safe based on the established guidelines around the world. However, due to their persistence and potential to accumulate and reach higher concentrations in the environment, careful monitoring of PFASs in all types of water is critical.

Carbon Monoxide in the Expired Air and Urinary Cotinine Levels of e-Cigarette Users.

OBJECTIVES: The aim of this descriptive study was to determine the sociodemographic characteristics of electronic (e)-cigarette users for clarifying the causes of e-cigarette smoking and to identify the carbon monoxide (CO) and urinary cotinine levels of the volunteers. MATERIALS AND METHODS: Twenty volunteers who smoked e-cigarettes completed a questionnaire, and their exhaled CO and urinary cotinine levels were measured. An enzyme-linked immunosorbent assay kit was used for cotinine analysis. RESULTS: Overall, 85% of the participants were males, 60% were married, and 75% were college/university graduates. The median age of participants was 38.5 years. The participants' main reasons for starting to smoke were peer influence and curiosity. The participants' main reasons for smoking e-cigarettes were to quit and reduce smoking the conventional cigarettes and cost effectiveness. Only three people knew that smoking was harmful to health. The participants' CO levels were measured as a median of 3, lowest of 1, and highest of 22. Cotinine levels were "positive" in all samples. A moderate and statistically significant correlation was found between the amount of fluids used by the participants in 1 day (mL) and cotinine levels in urine specimens (Pearson correlation test, r=0.511, p=0.025). CONCLUSION: The study is an important proof of the country's scientific work on e-cigarettes. Preventive strategies should be very strictly implemented for any tobacco products, including e-cigarettes, as they harm individuals and the community.

Ciprofloxacin-induces free radical production in rat cerebral microsomes.

In the presence of ciprofloxacin (CPFX), free radical adduct formation was demonstrated in rat cerebral microsomes using a spin trap α-(4-pyridyl-1-oxide)-N-tert-butyl-nitrone by electron spin resonance spectroscopy. Active microsomes, dihydronicotinamide-adenine dinucleotide phosphate, and ciprofloxacin were necessary for the formation of a spin trap/radical adduct. Adduct formation increased dose-dependently at 0.5-1 mM CPFX concentration for 180 min, and 0.3-1 mM concentration level for 240 min. The addition of SKF 525A, ZnCl2 or desferrioxamine to the incubation system caused complete inhibition of the radical formation. However, pretreatment of microsomal system with superoxide dismutase (SOD) did not induce any protective effect. Induction of lipid peroxidation, and depletion of thiol levels by CPFX were also shown in the system. These results strongly suggested that CPFX produces free radical(s) in the cerebral microsomes of rats.

Simple, fast and reliable liquid chromatographic and spectrophotometric methods for the determination of theophylline in urine, saliva and plasma samples.

In this study, a high-performance liquid chromatographic method (HPLC) and UV spectrophotometric method were developed, validated and applied for the determination of theophylline in biological fluids. Liquid- liquid extraction is performed for isolation of the drug and elimination of plasma and saliva interferences. Urine samples were applied without any extraction. The chromatographic separation was achieved on a C18 column by using 60:40 methanol:water as mobile phase under isocratic conditions at a flow rate of 0.75 mL/min with UV detection at 280 nm in HPLC method. UV spectrophotometric analysis was performed at 275 nm. THE RESULTS OF HPLC ANALYSIS WERE AS FOLLOWS: the limit of quantification: 1.1 µg/mL for urine, 1.9 µg/mL for saliva, 3.1 µg/mL for plasma; recovery: 94.85% for plasma, 100.45% for saliva, 101.39% for urine; intra-day precision: 0.22-2.33%, inter-day precision: 3.17-13.12%. Spectrophotometric analysis results were as follows: the limit of quantitation: 5.23 µg/mL for plasma, 8.7 µg/mL for urine; recovery: 98.27% for plasma, 95.25% for urine; intra-day precision: 2.37 - 3.00%, inter-day precision: 5.43-7.91%. It can be concluded that this validated HPLC method is easy, precise, accurate, sensitive and selective for determination of theophylline in biological samples. Also spectrophotometric analysis can be used where it can be applicable.

Toxic metals in breast milk samples from Ankara, Turkey: assessment of lead, cadmium, nickel, and arsenic levels.

Toxic metals are one of the significant groups of chemical contaminants that humans are exposed to by oral, inhalation, and dermal routes. Exposure to these chemicals begins with intrauterine life and continues during lactation period at the first years of life. Breastfeeding has a much more special place than other nutrition options for infants. However, when possibility of contaminant transfer by breast milk is considered, its safety and quality is essential. Regarding infant and mother health and limited number of information on this field in Turkey, measuring contamination levels in breast milk is important. Therefore, in the present study, lead (Pb), cadmium (Cd), nickel (Ni), and arsenic (As) levels were measured by atomic absorption spectrometry in 64 breast milk samples obtained from mothers from Ankara, Turkey. Pb and Ni levels in breast milk samples were found to be 391.45±269.01 µg/l and 43.94±33.82 µg/l (mean ± SD), respectively. Cd was found only in one of 64 samples, and the level was 4.62 µg/l. As level was below the limit of quantification (LOQ, 7.6 µg/l) in all samples. These findings will accurately direct strategies and solutions of protection against contaminants in order to reduce their levels in biological fluids.

Protective effect of zinc chloride against cobalt chloride-induced cytotoxicity on vero cells: preliminary results.

The aim of this study was to investigate the possible time- and dose-dependent cytotoxic effects of cobalt chloride on Vero cells. The cultured cells were incubated with different concentrations of cobalt chloride ranging from 0.5 to 1,000 µM, and cytotoxicity was determined by 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) and resazurin assays. Possible protective effects of vitamin E, coenzyme Q(10), and zinc chloride were also tested in this system. A gradual decrease in cell proliferation was observed at concentrations ~≥ 200 µM in incubation periods of 24, 48, 72, and 96 h with MTT assay. Exposure of cells to 500 and 1,000 µM cobalt chloride caused significant decrease in cell survival. A biphasic survival profile of cells was observed at 1-25 µM concentration range following 96 h of incubation. With resazurin assay, cytotoxicity profile of CoCl(2) was found comparable to the results of MTT assay, particularly at high concentrations and long incubation periods. Dose-dependent cytotoxicity was noted following exposure of cells to ≥ 250 µM of CoCl(2) for 24 h and ≥ 100 µM concentrations of CoCl(2) for 48-96 h. Pretreatment of cells with ZnCl(2) for 4 or 24 h provided significant protection against cobalt chloride-induced cytotoxicity when measured with MTT assay. However, vitamin E or coenzyme Q(10) was not protective. CoCl(2) had dose- and time-dependent cytotoxic effects in Vero cells. Preventive effect of ZnCl(2) against CoCl(2)-induced cytotoxicity should be considered in detail to define exact mechanism of toxicity in Vero cells.

Cytotoxic effect of ciprofloxacin in primary culture of rat astrocytes and protection by Vitamin E.

The aim of this study was to investigate the possible cytotoxic and oxidative stress inducing effects of ciprofloxacin (CPFX) on primary cultures of rat astrocytes. The cultured cells were incubated with various concentrations of CPFX (0.5-300mg/l), and cytotoxicity was determined by neutral red (NR) and MTT assays. Survival profile of cells was biphasic in NR assay: CPFX did not cause any alteration at any concentration for 7h, whereas < or =50mg/l concentrations induced significant cell proliferation in incubation periods of 24, 48, 72, and 96h. However, cell proliferation gradually decreased at higher concentrations, and 200 and 300mg/l of CPFX exposure was found to be significantly (p<0.05) cytotoxic at all time periods. With MTT assay, no alteration was noted for incubation period of 7h, as observed with NR assay. But, cell viability decreased with approximately > or =50mg/l CPFX exposure in all other time periods. Cell proliferation was only seen in 24h of incubation with 0.5 and 5mg/l CPFX. Vitamin E pretreatment of cell cultures were found to be providing complete protection against cytotoxicity of 300mg/l CPFX in 96h incubation when measured with both NR and MTT assays. The SOD pretreatment was partially protective with NR assay, but no protection was noted when measured with MTT. A significant enhancement of lipid peroxidation was observed with the cytotoxic concentration of the drug, but total glutathione content and catalase activity of cells did not change. The data obtained in this study suggest that, in accordance with our previous results with fibroblast cells, CPFX-induced cytotoxicity is related to oxidative stress. And the biphasic effect of CPFX possibly resulted from the complex dose-dependent relationships between reactive oxygen species, cell proliferation, and cell viability.

Ciprofloxacin-induced DNA damage in primary culture of rat astrocytes and protection by Vitamin E.

Fluoroquinolones are generally well-tolerated antibiotics in patients. Gastrointestinal, central nervous system, and dermatological adverse events were the most frequent unwanted effects during therapy with these drugs. However, the mechanism of these adverse effects has not yet been elucidated. The aim of this study was to investigate the possible DNA damage-inducing effect of a fluoroquinolone (FQ) antibiotic, ciprofloxacin (CPFX) on primary culture of rat astrocytes. For this purpose, the cultured cells were incubated with various concentrations of CPFX, and DNA damage was monitored by comet assay. Our results showed a concentration-dependent induction of DNA damage by CPFX. Pretreatment of cells with Vitamin E for 4h provided partial protection against this effect. The data obtained in this study suggest that CPFX-induced DNA damage might be related to oxidative stress and should be considered for further mechanistic studies of central nervous system toxicity of CPFX.

Ciprofloxacin-Induced Cytotoxicity and Apoptosis in HeLa Cells.

The aim of this study was to investigate the possible cytotoxic and apoptotic effects of a fluoroquinolone (FQ) antibiotic, ciprofloxacin (CPFX), on HeLa and HeLa-tat cell lines. The cultured cells were incubated with three different (20 to 100 mg/L) concentrations of CPFX, and cytotoxicity was determined by trypan blue test. A dose- and time-dependent decrease in cell proliferation was observed for both cell types except 50 to 100 mg/L CPFX for 24 h for HeLa-tat cells. CPFX-induced apoptosis was also measured by the acridin orange-ethidium bromide staining using a fluorescence microscope in both cell types. Our results showed an induction of apoptosis at 100 mg/L concentration of CPFX after 24 and 48 h of incubation in both cell types. These data confirmed our previous studies obtained with normal human fibroblast cell cultures and indicated that CPFX induces cytotoxicity and apoptosis in HeLa cells.

Ciprofloxacin-induced glutathione redox status alterations in rat tissues.

The possible oxidative stress inducing effect of a fluoroquinolone (FQ) antibiotic, ciprofloxacin (CPFX), was investigated in rats measuring glutathione redox status. For this purpose, the drug was administered to rats as two different single doses (100 and 150 mg/kg, ip) or a repeated dose (500 mg/kg/d, ig, for 5d). Then, total and oxidized glutathione levels were determined in hepatic and cerebral tissues of the rats by an enzymatic cycling assay, and the glutathione redox status was calculated. The possible protective effects of vitamin E or allopurinol against CPFX-induced alterations on glutathione system have also been examined. Following both routes of administration of CPFX, the total glutathione content of the liver, but not of brain decreased significantly. The oxidized glutathione (GSSG) in the brain increased after single or repeated dose treatments, but only with repeated doses of CPFX in the liver. CPFX induced dose-dependent alterations in the glutathione redox status in both tissues. With single doses the effect was more pronounced in cerebral tissue, and with repeated ig doses it was significant in both tissues. Pretreatment of rats with vitamin E or allopurinol before the administration of CPFX provided marked protection against glutathione redox status alterations in both tissues. Our results, thus, indicate that CPFX treatment introduces an oxidative stress in cerebral and hepatic tissues of rat.

Biphasic response of ciprofloxacin in human fibroblast cell cultures.

To investigate the possibility of the involvement of an oxidative stress induction in the mechanism of the cytotoxic effect of quinolone antibiotics, we examined the viability of human fibroblast cells exposed to ciprofloxacin (CPFX), and measured the levels of lipid peroxidation (LP), glutathione (GSH), and the activities of the antioxidant enzymes catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPX). The data showed that the effect of CPFX on the viability of cells, as determined by neutral red uptake assay, was time-dependent, and the dose-response relation was biphasic. Cytotoxicity was not observed in the concentration range 5-150 mg/l CPFX when the cells were incubated for 24 h. In contrast, lower concentrations (5 and 12.5 mg/l) of CPFX increased the cell growth in all incubation periods tested. Marked decreases in the viability of fibroblasts were observed at concentrations 50 and 75 mg/l, and >/=50 mg/l, following 48 and 72 h exposure, respectively (p < 0.05). However, when the cells were exposed to > 75 mg/l CPFX for 48 h, no cytotoxicity was observed. By exposing fibroblast cultures to 75 mg/l CPFX for 48 h, an induction of LP enhancement and a marked decrease in intracellular GSH were observed. Vitamin E pretreatment of the cells lowered the level of LP, increased the total GSH content, and provided significant protection against CPFX-induced cytotoxicity. The biphasic effect of CPFX possibly resulted from the complex dose-dependent relationships between reactive oxygen species (ROS), cell proliferation, and cell viability. It was previously reported, in fact, for several cell models that ROS exert a biphasic effect on cell growth. Furthermore, cultured fibroblasts release their own free radicals, and the inhibition of endogenous ROS inhibits the fibroblast cell proliferation, whereas the effect of exogenous ROS is biphasic.