Search

The need to create safe materials for biomedical and pharmaceutical applications has become a significant driving force for the development of new systems. Therefore, a chitosan-coated copolymer of itaconic acid, acrylic acid, and N-vinyl caprolactam (IT-AA-NVC) was prepared by radical polymerization and subsequent coating via nanoprecipitation to give a system capable of sustained delivery of melatonin. Although melatonin brings undoubted benefits to the human body, aspects of the optimal dose, route, and time of administration for the obtaining of suitable treatment outcomes remain under discussion. The entrapment of melatonin in biocompatible polymeric systems can prevent its oxidation, decrease its toxicity, and provide an increased half-life, resulting in an enhanced pharmacokinetic profile with improved patient compliance. The structures of the biopolymer and conjugate were proven by FTIR, thermal properties were tested by DSC, and the morphologies were followed by SEM. The loading efficiency and in vitro release profile were studied by means of HPLC, and a delayed release profile with an initial burst was obtained. The potential systemic toxicity of the formulation was studied in vivo; a mild hepatotoxicity was observed following administration of the melatonin-loaded formulation to mice, both by histopathology and blood clinical biochemistry. Histopathology showed a mild nephrotoxicity as well; however, kidney clinical biochemistry did not support this. [ABSTRACT FROM AUTHOR]

Concentrations of persistent organic pollutants (POPs) were quantified in river water and sediment, as well as in the liver and muscle tissues of Cyprinus carpio that were sampled four times in a year at three stations in the Büyük Menderes River (BMR). Potential biomarkers of possible cellular molecular damage, namely lipid peroxidation (LPO) degradation products, protein carbonyls (PCO) and DNA repair product 8-hydroxy-2′-deoxyguanosine (8-OHdG), were analysed. All the targeted pollutants were measurable both in biotic and abiotic samples. Interestingly, the results suggested that there was recent organochlorine pesticide (OCP) input into the river water in the first two sampling periods in all stations in contrast to prohibition, while input of polychlorinated biphenyls (PCBs) and polybrominated diphenylethers (PBDEs) was not detected. Liver POP concentrations were higher than in muscle, as expected, and were found to decrease from the first to the fourth sampling period in all stations, except PBDEs. Levels of LPO degradation products in the liver and in muscle tissues decreased from the first to the fourth sampling period. This suggests that these markers reflect the lipid damage in respective tissues due to the tissue burden of targeted POPs. Protein carbonyls were the highest in the first sampling period, followed by a dramatic decrease in the second, and then a gradual increase towards the fourth sampling period in all stations. 8-OHdG levels were lower in Sarayköy station in the first sampling period. Among the measured biomarkers, only several LPO degradation products were significantly correlated with OCPs and PCBs in liver tissue. [ABSTRACT FROM AUTHOR]

The Büyük Menderes River (BMR) is one of the largest rivers in Turkey. This river irrigates efficient farmlands and includes tributaries of other rivers and streams and many populated towns within its limits in the Ege region. Both the estuary and Işıklı Lake serve as a sanctuary for various waterbirds. Therefore, the BMR plays a critical role both for the inhabitants and for the ecosystem organisms in its environs. In the present study, we analyzed levels of metals including iron, barium, zinc, vanadium, cobalt, chromium, cadmium, copper, nickel, aluminum, arsenic, manganese, antimony, silver, selenium, boron, mercury, titanium, and lead in river water, sediment, fish ( Cyprinus carpio; common carp), and in various waterbird ( Fulica atra, Euroasian coot; Larus michahellis, yellow-legged gull; Ardea cinerea, grey heron; Larus melanocephalus, Mediterranean gull; and Pelecanus crispus, pelican) samples. Analyses were performed using an inductively coupled plasma-mass spectrometry (ICP-MS) instrument after sample preparation. Comparing metal concentrations among different sample types, it was found that barium, aluminum, and zinc are the major metals in river water, and zinc in common carp muscle, while iron, aluminum, and manganese are the major metals in sediments. Iron, zinc, copper, and aluminum were the highest in waterbird muscle tissue. Iron and barium were found to be the major metals in eggshell, while iron and zinc are the major metals in egg samples. A simple 'worst-case scenario' model of risk assessment revealed that some of the analyzed metals may pose a risk for human health through consuming fish. [ABSTRACT FROM AUTHOR]

ELISA is commonly used for the detection of urinary 8-oxo-7,8-dihydro-2′-deoxyguanosine (8-oxodG), a marker of whole body oxidative stress. However, the method has been criticized for high inter-laboratory variability and poor agreement with chromatographic techniques. We performed an inter-laboratory comparison of 8-oxodG assessed in 30 urine samples and a urine spiked with four different concentrations of 8-oxodG by ELISA using standardized experimental conditions, including: sample pre-treatment with solid-phase extraction (SPE), performing analysis using a commercial kit from a single manufacturer and strict temperature control during the assay. We further compared the ELISA results with high-performance liquid chromatography-tandem mass spectrometry (HPLC–MS/MS) and performed tentative identification of compounds that may contribute to the discrepancy between both methods. For all but one participating laboratory (Data 1) we observed consistent ELISA results lying mostly within 1 SD of the mean 8-oxodG concentration. Mean 8-oxodG levels assessed by ELISA correlated with the data obtained by HPLC–MS/MS ( R =0.679, p <0.001). The correlation improved when Data 1 were excluded from the analysis ( R =0.749, p <0.001). We identified three outlying urine samples; one with an ELISA 8-oxodG concentration lower, and two with 8-oxodG levels higher, than those measured by HPLC–MS/MS. Omitting these samples further improved inter-methodology agreement ( R =0.869, p <0.001). In the outliers with high 8-oxodG estimates various aromatic and heterocyclic compounds were tentatively identified using gas chromatography-mass spectrometry (GC–MS). Application of authentic standards revealed the presence of saccharides, including d -glucose and d -galactose as putative interfering substances. In summary, assay standardization improved ELISA inter-laboratory agreement, although some variability is still observed. There are still compounds contributing to overestimation of 8-oxodG by ELISA, but only in some urine samples. Thus, despite significant improvement, ELISA still should not be considered a robust alternative to chromatographic techniques. [ABSTRACT FROM AUTHOR]

We explored the association between the activities of antioxidant enzymes and their metallic cofactors in rats treated with cisplatin. The antioxidant effects of aminoguanidine, and a combination of vitamins E and C were investigated. Plasma platin was significantly lower than liver and kidney. Cisplatin treatment caused significant increase in plasma Se-glutathione peroxidase activity. Activities of Se-glutathione peroxidase, glutathione S-transferase, catalase and Cu,Zn-superoxide dismutase have been found to be significantly decreased in liver and kidney compared to controls. Zn levels in these organs were diminished upon cisplatin treatment, while levels of Cu were unaffected. Interestingly, levels of iron, the cofactor of catalase, were found to be significantly increased in liver and kidney. Intervention with aminoguanidine or vitamins was generally prevented cisplatin-caused changes in the activity of enzymes and in the tissue levels of cofactor metals. These observations suggest that relation between activities of enzymes and levels of cofactor metals is multifactorial. [ABSTRACT FROM PUBLISHER]

Objective: To investigate whether lipid and protein oxidation products are elevated and correlated with routine clinical markers of hepatic and renal function in patients anesthetized with halothane, isoflurane, or sevoflurane. Methods: Urine and blood samples were collected from patient groups. Excretion of aldehydes, acetone, and o,o'-dityrosine was measured before and after anesthesia. Blood samples were analysed for clinical markers. Results: Urinary concentrations of aldehydes, acetone, o,o'-dityrosine and glucose were significantly increased after anesthesia in halothane and sevoflurane groups earlier than clinical markers. Significant correlations were found in sevoflurane group. Conclusion: Lipid and protein oxidation contributes to subclinical sevoflurane nephrotoxicity. Oxidation products may serve as early biomarkers. [ABSTRACT FROM AUTHOR]

New, except 1d, melatonin analogue benzimidazole derivatives were synthesized and characterized in the present study. The potential role of melatonin as an antioxidant by scavenging and detoxifying ROS raised the possibility that compounds that are analogous to melatonin can also be used for their antioxidant properties. Therefore the antioxidant effects of the newly synthesized compounds were investigated in vitro by means of their inhibitory effect on hydrogen peroxide-induced erythrocyte membrane lipid peroxidation (EMLP) and on various erythrocyte antioxidant enzymes viz. superoxide dismutase (SOD), catalase (CAT) and glucose-6-phosphate dehydrogenase (G6PD). The synthesized benzimidazole derivatives showed remarkable antioxidant activity in vitro in the H 2 O 2 -induced EMLP system. Furthermore their effects on various antioxidant enzymes are discussed and evaluated from the perspective of structure- activity relationships. [ABSTRACT FROM AUTHOR]

Environmental pollutants such as organochlorine pesticides (OCPs), polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs), so called persistent organic pollutants (POPs) have been known to interfere various ion channels in organism, which is the main mechanism of action especially for OCPs. Furthermore, all PCBs and several congeners of OCPs and PBDEs were recently listed as "carcinogen" by the IARC depending on evaluation of epidemiological studies. On the other hand, there have been very little data on the relation between various cancers and tissue burden of these toxicants. In majority of the human studies on a possible association between POPs and cancers, exposure data have been based on statement-based interviews, which potentially impair the accuracy and reliability of quantitative assessment. In order to accurately assess whether environmental exposure to POPs increase risk of kidney, stomach and breast cancers, tumour tissues as well as blood and urines were collected from surgically operated patients. Cellular DNA and protein oxidative damage markers (8-OHdG and dityrosine, respectively), have also been analysed in the patient and healthy control groups, and assessed whether there are changes in these parameters. The data suggest that majority of POPs in cancerous tissues were associated with cancers in patients, while they are less conclusive in blood of the same patients. DNA and protein damage markers were found to be higher in urines of kidney cancer patients compared to healthy volunteers, although inter-individual variability obscured statistical significances. Current findings confirmed that glutathione S-transferase theta1 (GSTT1) nullpolymorphism is a risk factor Bforro ukgidhnte tyo cyaonuc ebry, |a nEdP cIyptsowchicrohme P450 1A1 (CYP1A1) is a risk factor for stomach cancer. Data suggest that environmental exposure to POPs may induce tumour initiation and/or progression in humans however, blood concentrations of the same pollutants do not adequately reflect tissue concentrations except dichlorodiphenyldichloroethylene (DDE), the major metabolite of the notorious organochlorine pesticide dichlorodiphenyltrichloroethylene (DDT). Possible role of interaction of POPs with ion channels in cancer initiation and/or progression remain to be explored. [ABSTRACT FROM AUTHOR]

Cigarette smoking leads to uptake of a multitude of reactive chemicals including many electrophiles and may also give rise to oxidative stress. Human red blood cells are important targets for electrophilic and oxidant foreign compounds. We investigated the oxidative stress in erythrocytes upon cigarette smoking, and the response of antioxidant defense system against it. With this aim, simultaneous determination of erythrocyte superoxide dismutase (SOD), selenium dependent glutathione peroxidase (Se-GPx), catalase (CAT), glutathione S-transferase (GST) activities and plasma levels of thiobarbituric acid reactive substances (TBARS), and the degree of erythrocyte membrane lipid peroxidation (EMLP) were carried out in blood samples of smokers and their controls. Plasma TBARS levels and EMLP in smokers were significantly higher than the control levels ( p < 0.01 and p < 0.005, respectively). SOD activity was diminished in smokers compared to nonsmoker controls ( p < 0.005). Erythrocyte Se-GPx activity was also found significantly diminished in smokers ( p < 0.005), while plasma Se-GPx activity was not changed. We observed that erythrocyte CAT activity was not different in smokers compared to nonsmoker controls. We found that the erythrocyte GST activity is significantly lower in young adult smokers (3.03 ± 0.18 U/mg protein; mean ± SEM; n = 46) than in nonsmoking contemporaries (3.98 ± 0.26 U/mg protein; mean ± SEM; n = 41). Together with previously reported data, it can be concluded that the decrease in GST activity leads to extra GST synthesis during erythrocyte proliferation. The same data were also analyzed for the sex differences. The statistically significant differences remained the same between nonsmoker and smoker females. Only EMLP degree and SOD activity were significantly different between nonsmoker and smoker males; however, when compared the parameters between male and female nonsmokers, GST activity was found to be significantly higher in females than that of males. © 2005 Wiley Periodicals, Inc. J Biochem Mol Toxicol 19:226-233, 2005; Published online in Wiley InterScience (). DOI 10.1002/jbt.20088 [ABSTRACT FROM AUTHOR]

The objective of the present study was to evaluate a comprehensive set of urinary biomarkers for oxidative damage to lipids, proteins and DNA, in man. Eighteen moderately trained males (mean age 24.6±0.7) exercised 60 min at 70% of maximal O 2 uptake on a cycle ergometer. Urine fractions for 12 h were collected 1 day before, and for 3 consecutive days after exercise. As biomarkers of lipid peroxidation, 8 aldehydes (i.e. propanal, butanal, pentanal, hexanal, heptanal, octanal, nonanal and malondialdehyde--MDA)and acetone were analyzed in urines by gas chromatography with electron capture detection (GC-ECD). As a biomarker of protein oxidation, o , o ′-dityrosine was analyzed in urine samples by a recently developed isotope dilution HPLC-atmospheric pressure chemical ionization (APCI)-tandem-mass spectrometry (HPLC-APCI-MS/MS) methodology. As a biomarker of oxidative DNA damage, urinary excretion of 8-hydroxy-2′-deoxyguanosine (8-OHdG) was measured by an ELISA method. On the day of exercise, significant increases were observed in urinary excretions of acetone ( p <0.025, n =18 ) and butanal ( p <0.01, n =18 ) in the 12 h daytime fractions compared to the daytime fraction before exercise. The urinary acetone excretion was also significantly ( p <0.05) increased on the 1st day after exercise. Octanal and nonanal were increased in the daytime urine fraction on the 2nd day after exercise. However, these increases were of borderline significance ( p =0.09 and p =0.07, respectively). Significantly elevated urinary o , o ′-dityrosine amounts were observed in the daytime fraction on the day of exercise ( p <0.025) and on the 1st day after exercise ( p =0.07) compared to the before exercise daytime fraction. Excretion of urinary 8-OHdG was statistically significantly increased in the daytime fractions on the day of exercise ( p =0.07) and on the 1st day after exercise ( p <0.025) compared to before exercise daytime fraction. Increases in urinary excretions of acetone, propanal, pentanal, MDA and 8-OHdG significantly correlated with training status (hours of exercise/week) of the volunteers, while o , o ′-dityrosine did not. To our knowledge, the present study is the first to evaluate a multi-parameter non-invasive biomarker set for damage to three main cellular targets of ROS. It shows that 1 h of exercise may already induce oxidative damage in moderately trained individuals and that the chosen urinary biomarkers are sensitive enough to monitor such damage. [ABSTRACT FROM AUTHOR]

Acetaminophen (APAP) hepatotoxicity is the most common cause of acute liver failure in the United States, and while a significant percentage of APAP overdose patients develop kidney injury, molecular mechanisms involved in APAP-induced nephrotoxicity are relatively unknown. We have shown that 4-methylpyrazole (4MP, Fomepizole) protects against APAP-induced liver injury by inhibiting reactive metabolite formation through Cyp2E1, and analysis of data from APAP overdose patients indicated that kidney dysfunction strongly correlated with severe liver injury. Since Cyp2E1 is also expressed in the kidney, this study explored protection by 4MP against APAP-induced nephrotoxicity. Male C57BL/6 J mice were treated with either 300 or 600 mg/kg APAP with or without 4MP for 2, 6 or 24 h, followed by measurement of APAP metabolism and tissue injury. Interestingly, levels of APAP and its non-oxidative metabolites were significantly higher in kidneys when compared to the liver. APAP-protein adducts were present in both tissues within 2 h, but were absent in kidney mitochondria, unlike in the liver. While GSH depletion was seen in both tissues, activation of c-jun N-terminal kinase and its translocation to the mitochondria, which is a critical feature of APAP-induced liver injury, was not detected in the kidney. Treatment with 4MP attenuated APAP oxidative metabolite generation, GSH depletion as well as kidney injury indicating its potential use in protection against APAP-induced nephrotoxicity. In conclusion, since reactive metabolite formation seems to be common in both liver and kidney, 4MP mediated inhibition of Cyp2E1 protects against APAP-induced nephrotoxicity. However, downstream mechanisms of APAP-induced nephrotoxicity seem distinct from the liver. • APAP causes renal failure in 12% of overdose patients with severe liver injury. • APAP overdose causes liver injury in mice and renal damage only at higher doses. • Cyp2E1 levels and APAP protein adducts are higher in liver compared to kidney. • In kidney there was no mitochondrial protein adducts or JNK activation detectable. • 4-methylpyrazole prevented APAP-induced kidney injury by inhibiting Cyp2E1. [ABSTRACT FROM AUTHOR]

Paracetamol (PAR) overdose is associated with massive hepatic injury; it may induce kidney toxicity as well. It is essential to measure organ-specific activities of related CYPs for evaluating the overdose cases. Available HPLC-based methods require high amounts of tissue samples. In order to develop liquid chromatography mass spectrometry (LC–MS)-based methods to process small amounts of human tissues, liver and kidney samples were obtained. Individual microsomes were prepared and incubated with PAR (for quantifying bioactivation), with nifedipine (for measuring CYP3A4 activity) and with p -nitrophenol (for measuring CYP2E1 activity). The small amount of tissue microsomes was sufficient to measure both the formation of NAPQI and the activities of CYP enzymes. Although the sample size in group was relatively low, both NAPQI formation and activity of CYP2E1 were significantly higher in males compared to females in kidney. Considerable variations in the metabolic capacity of individuals were observed for both organs. [ABSTRACT FROM AUTHOR]

Aim: To provide in vitro data on toxicity mechanisms of clozapine, diclofenac and nifedipine., Background: CHO-K1 cells were used as in vitro model to explore mechanisms of cytotoxicity of the test drugs., Objective: Cytotoxic mechanisms of clozapine (CLZ), diclofenac (DIC) and nifedipine (NIF) were studied in CHO-K1 cells in vitro. All three drugs induce adverse reactions in some patients with partially unknown mechanisms., Method: Following the determination of time- and dose-dependency of cytotoxicity by the MTT test, cytoplasmic membrane integrity was explored by the LDH leakage test. Both end-points were further examined in the presence of soft and hard nucleophilic agents, glutathione (GSH) and potassium cyanide (KCN), respectively, and either individual or general cytochrome P450 (CYP) inhibitors, whether CYP-catalysed formation of electrophilic metabolites play a role in the observed cytotoxicity and membrane damage. The generation of reactive metabolites during the incubations was also explored. Formation of malondialdehyde (MDA) and oxidation of dihydrofluorescein (DCFH) were monitored whether peroxidative membrane damage and oxidative stress take place in cytotoxicity. Incubations were also conducted in the presence of chelating agents of EDTA or DTPA to explore any possible role of metals in cytotoxicity by facilitating electron transfer in redox reactions. Finally, mitochondrial membrane oxidative degradation and permeability transition pore (mPTP) induction by the drugs were tested as markers of mitochondrial damage., Results: The presence of an individual or combined nucleophilic agents significantly diminished CLZ- and NIF-induced cytotoxicities, while the presence of both agents paradoxically increased DIC-induced cytotoxicity by a factor of three with the reason remaining unknown. The presence of GSH significantly increased DIC-induced membrane damage too. Prevention of membrane damage by the hard nucleophile KCN suggests the generation of a hard electrophile upon DIC and GSH interaction. The presence of CYP2C9 inhibitor sulfaphenazol significantly diminished DIC-induced cytotoxicity, probably by preventing the formation of 4-hydroxylated metabolite of DIC, which further converts to an electrophilic reactive intermediate. Among the chelating agents, EDTA caused a marginal decrease in CLZ-induced cytotoxicity, while DIC-induced cytotoxicity was amplified by a factor of five. Both reactive and stable metabolites of CLZ could be detected in the incubation medium of CLZ with CHO-K1 cells, which are known to have low metabolic capacity. All three drugs caused a significant increase in cytoplasmic oxidative stress by means of DCFH oxidation, which was confirmed by increased MDA from cytoplasmic as well as mitochondrial membranes. The addition of GSH paradoxically and significantly increased DIC-induced MDA formation, in parallel with the increase in membrane damage when DIC and GSH combined., Conclusion: Our results suggested that the soft electrophilic nitrenium ion of CLZ is not responsible for the observed in vitro toxicities, and this may originate from a relatively low amount of the metabolite due to the low metabolic capacity of CHO-K1. A hard electrophilic intermediate may contribute to cellular membrane damage incubated with DIC, while a soft electrophilic intermediate seems to exacerbate cell death by a mechanism other than membrane damage. A significant decrease in cytotoxicity of NIF by GSH and KCN suggested that both soft and hard electrophiles contribute to NIF-induced cytotoxicity. All three drugs induced peroxidative cytoplasmic membrane damage, while only DIC and NIF induced peroxidative mitochondrial membrane damage, which suggested mitochondrial processes may contribute to adverse effects of these drugs in vivo., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

In vivo biotransformation of exposed chemicals is one of the major factors that determine the concentration and the duration of a substance at the systemic site of effect. Given that toxicity is expressed as a function of two factors, namely dose and time, the type and intensity of the toxicity are directly dependent on the chemical transformation of the exposed parent substance. This dependency involves two different situations. The amount of the chemical reaching the target will be decreased with the extent of metabolism if the parent chemical is toxic, and the opposite is true if the metabolite(s) is toxic instead. To date, the liver microsomal fraction in mammals has been justifiably considered as the center of biotransformation reactions because the liver and microsomes (i.e., endoplasmic reticulum component of the cell) possess the most abundant types and quantities of xenobiotic-metabolizing enzymes, especially the cytochrome P450 supergene enzyme family. These enzymes are common in all kingdoms of life, which strongly suggests that the origin of life is common. It is already known that various drugs enter mitochondria by different mechanisms, and this translocation is believed to be responsible for mitochondrial effects that are part of the therapeutic actions of various drugs such as lipid-lowering statins or antidiabetogenic thiazolidindiones. However, the discovery of mitochondrial forms of the xenobiotic-metabolizing enzymes provoked discussions about whether mitochondria metabolize drugs and other chemicals to some extent. This possibility may particularly be important as mitochondria have various critical cellular structures and functions. In the case of in situ generated metabolite(s), when there are adverse interactions with either these structures or functions, various toxic outcomes may appear. In this review, we compiled studies in the literature regarding biotransformation of drugs and other chemicals catalyzed by mitochondria where it is both an initiator and target of toxicity., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)