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Graphical abstract, Highlights • Public consultation to update CLP regulation has launched. • Envisaged revisions will go beyond the classification and labelling of chemicals. • Endocrine disruptors and two categories of persistent toxic chemicals are proposed. • Main domains are environmental contamination, chemical mixtures and laboratory animals use., Recent advancements in toxicology and the European Union’s Green Deal, with its Chemicals Strategy for Sustainability, have paved the way for major changes in EU legislation on the control of environmental chemicals for a cleaner and safer environment. Another substantial legislative advancement underway is the update of the “Regulation on Classification, Labelling and Packaging of Substances and Mixtures (CLP),” an ambitious piece of EU legislation with exceptional scientific toxicological background in identifying a hazard, aiming at better protecting its citizens and the environment from the risk of chemical substances and products, the occupational settings included. Update of CLP legislation additionally aims at facilitating the free exchange of chemicals in the European Internal Market, provided that proper labelling and packaging processes are implemented. Participation in the ongoing online public consultation on these issues, ending on November 15, 2021, is of key relevance to ensure a transparent and effective definition of such an important piece of legislation, fully compliant with current EU priorities in terms of human and environmental protection and animal welfare.

Acrylamide is a chemical, often present in bread, legally classified as carcinogen, mutagen and reproductive toxicant. Since bread is consumed both world-wide and in Iran, determination of acrylamide in different types of breads is of high interest. In the present study, acrylamide was monitored in 56 Sangak and 30 industrial bread samples collected from Tehran and Shiraz, using LC-MS/MS (LOQ = 1 ng/g). In addition, the noncarcinogenic risk (target hazard quotient–THQ) and carcinogenic risk (incremental lifetime cancer risk–ILCR) due to ingestion of acrylamide through bread consumption in children and adults were assessed. Acrylamide was detected in more than 90% of the samples tested. The average daily intake of acrylamide in Iran based on exclusive consumption of Sangak bread, was estimated at 145 ng/kg bw/day. Based on the THQ for bread acrylamide in adults and children, the decreasing risk order was: Shiraz semi-industrial Sangak, Shiraz traditional Sangak, Tehran traditional Sangak, Tehran industrial bread. The ILCR of bread acrylamide calculated for adults and children was higher than the permissible lifetime carcinogenic risk value established by USEPA (1.00E-5). Results show that bread is a major source of acrylamide intake by people in Iran and all consumers regardless of age could be at elevated carcinogenic risk.

The aim of this study was to identify organic components eluted from five resin dental sealants using gas chromatography and mass spectrometry (GC/MS) after 1-day and 40-days storage and the effect of sealants on cell survival of cultured fibroblasts. Five resin materials were studied: BeautiSealant (SHOFU), Clinpro (3M/ESPE), Conseal F (SDI), Grandio Seal (VOCO) and Helioseal Clear (Ivoclar/Vivadent). The organic monomers detected were butylated hydroxytoluene (BHT), bis-phenol-A (BPA), camphoroquinone (CQ), diethylenglycoldimethacrylate (DEGDMA), 4N, N-dimethylaminobenzoic acid butylethoxyester (DMABEE), hydroxyethylmethcrylate (HEMA), hydroquinone monomethylether (MEHQ), triethylene glycol dimethacrylate (TEGDMA), tetrabutylammonium tetrafluoroborate (TBATFB), triphenylstibane (TPSb). The main monomer detected was TEGDMA, whereas BHT and DEGDMA were detected at lower concentrations. Higher monomer concentrations were detected after 40 days storage. The eluting chemical profiles of the tested materials differ qualitative and quantitative. For cytotoxicity evaluation, NIH/3T3 cells were exposed to eluates of sealants and cell viability was assessed by a quantitative technique at two observation periods. Decreased cell viability was observed. The cytotoxicity and the release of monomers from dental materials examined depends on the type of material and the observation time point. Resin-based dental materials have raised public concerns regarding possible adverse biological effects, thus it is essential to evaluate possible side effects for human health.

Cardiovascular diseases are among the most significant causes of mortality in humans. Pesticides toxicity and risk for human health are controlled at a European level through a well-developed regulatory network, but cardiotoxicity is not described as a separate hazard class. Specific classification criteria should be developed within the frame of Regulation (EC) No 1272/2008 in order to classify chemicals as cardiotoxic, if applicable to avoid long-term cardiovascular complications. The aim of this study was to review the cardiac pathology and function impairment due to exposure to pesticides (i.e. organophosphates, organothiophisphates, organochlorines, carbamates, pyrethroids, dipyridyl herbicides, triazoles, triazines) based on both animal and human data. The majority of human data on cardiotoxicity of pesticides come from poisoning cases and epidemiological data. Several cardiovascular complications have been reported in animal models including electrocardiogram abnormalities, myocardial infarction, impaired systolic and diastolic performance, functional remodeling and histopathological findings, such as haemorrhage, vacuolisation, signs of apoptosis and degeneration.

Although the use of nutritional supplements by adult athletes has been extensively studied, information on supplements consumption by adolescent athletes is still limited. The present study reports on the use of nutritional supplements contaminated with banned doping substances among 170 recreational adolescent athletes from eleven, randomly selected, gym centres, in Athens, Greece. Nutritional supplements consumption was reported by almost 60% of the study population, with proteins/amino acids and vitamins being the most popular. Nine per cent of the users were found to consume nutritional supplements contaminated with anabolic steroids, prohormones, selective androgen receptor modulators (SARMs) and aromatase inhibitors, all pharmacological substances with endocrine modulating properties not stated on the label. None of these individuals had previously consulted a physician or a nutritionist. A representative sample (ca 15%) of the protein/aminoacids and creatine preparations used by the study population were also tested and found free from doping substances. The majority (63%) of adolescents purchased products from the internet. In conclusion, exercising adolescents can have easy access to contaminated nutritional supplements and "black market" products, which could constitute a risk for public health. Low level of awareness and low involvement of medical care professionals among recreational adolescent athletes is also observed.

Contrast-induced nephropathy (CIN) is reversible acute renal failure observed following administration of iodinated contrast media (CM) during angiographic or other medical procedures such as urography. There are various mechanisms through which CM develop their nephrotoxic effects, including oxidative stress and apoptosis. CIN is a real-life, albeit not very rare, entity. Exact pathophysiology remains obscure and no standard diagnostic criteria apply. The Acute Kidney Injury Network criteria was recently employed but its incidence/clinical significance warrants further clarification based on recent methodological advancements, because most published studies to date were contaminated by bias. The current study is a comprehensive review conducted to provide an overview of the basic concepts of CIN and summarize recent knowledge on its pathophysiology and the evidence supporting potential prevention strategies. CIN is expected to increase morbidity, hospital stay and mortality, while all patients scheduled to receive CM should undergo risk assessment for CIN and high-risk patients may be considered candidates for prevention strategies. The value of using compounds with antioxidant properties other than sodium bicarbonate, remains controversial, warranting further clinical investigation.

Contrast-induced nephropathy (CIN) is a leading cause of hospital-acquired acute kidney injury as a result of iodinated contrast-media use for diagnostic purposes. Pathophysiology remains unclear. In the present study iopromide was administered to New Zealand white rabbits without any prior intervention. Oxidative stress was assessed in blood and tissue level at three anatomical kidney areas (medullary, cortical, juxtamedullary). Histopathological evaluation was also performed. Serum creatinine and urea increased in the CIN groups over 25% at two hours after administration and returned to baseline at 48 h. In kidney tissues, a significant reduction (40%) of catalase in renal cortexes of the CIN groups was observed. Necrosis and tubular vacuolization was also noted that correlated with urea and creatinine levels. Lipid peroxidation decreased at 10 h after administration (>45%) and remained low even at 48 h. Plasma protein carbonyls were significantly increased (67%) in 2 h and dropped later. Serum levels of creatinine and urea at 24 and 48 h significantly correlated with the Total Antioxidant Activity and lipid peroxidation, respectively. Oxidative stress is shown to be involved in CIN development in the rabbit, with more pronounced effects to be confined to the cortex and outer stripe of the outer medulla.

The substance 4-methylimidazole (4-MEI) has raised several concerns regarding its toxicity to humans, although no harmonized classification has yet been decided. The regulatory limits for food products set by various authorities in Europe and the USA differ considerably. The purpose of the present study is to compare two liquid chromatography techniques in order to determine the levels of 4-MEI in food products from the Greek market and roughly estimate the possible exposure and relevant health risk for the consumers. A total of thirty-four samples (soft drinks, beers, balsamic vinegars, energy drinks and sauces) were collected and analyzed. The quality parameters for both analytical methodologies (linearity, accuracy, inter day precision, recovery) are presented. No detectable levels of 4-MEI are found in beers and soft drink samples, other than cola type. On the other hand, 4-MEI was detected in all cola type soft drinks (15.8–477.0 ng/ml), energy drinks (57.1%, 6.6–22.5 ng/ml) and vinegar samples (66.7%, 9.7–3034.7 ng/ml), while only one of the sauce samples was found to have a detectable level of 17.5 ng/ml 4-MEI.

Graphical abstract, Highlights • Organochlorine compounds (OCs) are persistent organic pollutants acting as endocrine disruptors. • Exposure to OCs is a risk factor for several severe pathologies. • Specific CYP polymorphisms could affect the clinical impact of OCs exposure., The association between genetic variations in the cytochrome P450 (CYP) family genes and pathological conditions related to long-term exposure to organochlorine compounds (OCs) deserves further elucidation. OCs are persistent organic pollutants with bioaccumulative and lipophilic characteristics. They can act as endocrine disruptors and perturb cellular mechanisms. Prolonged exposure to OCs has been associated with different pathological manifestations. CYP genes are responsible for transcribing enzymes essential in xenobiotic metabolism. Therefore, polymorphisms in these genetic sequences a. alter the metabolic pathways, b. induce false cellular responses, and c. may provoke pathological conditions. The main aim of this review is to define the interaction between parameters a, b and c at a mechanistic/molecular level, with references in clinical cases.

Cypermethrin (CY) is a frequently used class II pyrethroid pesticide, while piperonyl butoxide (PBO) plays a major role in the pesticide formulation of synthetic pyrethroids. Synthetic pyrethroids are metabolized in mammals via oxidation and ester hydrolysis. PBO can prevent the metabolism of CY and enhances its pesticide effect. While this potentiation effect reduces the amount of pesticide required to eliminate insects, it is not clear how this mixture affects mammals. In our in vivo experiment, New Zealand white male rabbits were exposed to low and high doses of CY, PBO, and their combinations, for 4 months. Genotoxicity and cytotoxicity were monitored by measuring binucleated cells with micronuclei (BNMN), micronuclei (MN) and the cytokinesis block proliferation index (CBPI) in lymphocytes. After two months of exposure, a statistically significant increase in the frequency of BNMN was observed for all exposed animals (p

3-monochloropropane-1,2-diol (3-MCPD) is a food contaminant that occurs during industrial production processes and can be found mainly in fat and salt containing products. 3-MCPD has exhibited mutagenic activity in vitro but not in vivo, however, a genotoxic mechanism for the occurrence of kidney tumors has not so far been excluded. The main pathway of mammalian 3-MCPD metabolism is via the formation of ß - chlorolactatic acid and formation of glycidol has been demonstrated in bacterial metabolism. The aim of this study was to investigate genotoxic and oxidative DNA damaging effects of 3-MCPD and its metabolites, and to provide a better understanding of their roles in DNA repair processes. DNA damage was assessed by alkaline comet assay in target rat kidney epithelial cell lines (NRK-52E) and human embryonic kidney cells (HEK-293). Purine and pyrimidine base damage, H2O2 sensitivity and DNA repair capacity were assessed via modified comet assay. The results revealed in vitro evidence for increased genotoxicity and H2O2 sensitivity. No association was found between oxidative DNA damage and DNA repair capacity with the exception of glycidol treatment at 20 µg/mL. These findings provide further insights into the mechanisms underlying the in vitro genotoxic potential of 3-MCPD and metabolites. © 2016 Elsevier Ltd. 36089/UDP-45005 This study was supported by Istanbul University Scientific Research Project ( 36089/UDP-45005 ). The authors wish to thank Dr. Ela Kadioglu (Gazi University, Turkey) and Dr. Andrew Collins (University of Oslo, Oslo) for kindly providing Endo III and Fpg enzymes to be used in the modified comet assay.

The aimof this study is to clarify the effect of cypermethrin (CY) on the oxidative stress (OS) and inflammation status of animals exposed to it and the synergistic role of piperonyl butoxide (PB0).Markers of oxidative stress, such as total antioxidant activity (TAC), protein carbonyls, hemoglobin (Hb), reduced glutathione (GSH), thiobarbituric-acid reactive substances (TBARS), along with the telomerase activity in PBMCs (peripheral blood mononuclear cells) were analyzed.Oxidative stress markers showed statistically significant differences between groups in TAC (p b 0.001), GSH (p = 0.018) and CAT activity (p = 0.029), which depended on dose and combined effect of both compounds. Telomerase activity also showed a statistically significant difference between all groups (F = 43.48, df=6, 14, p b 0.001)with cypermethrin, piperonyl butoxide and the co-exposed groups being significantly different fromthe control group (p b 0.05). Significance: The observed results for TBARS, GSH, Hb, TAC, Crbnls and CAT from our exposed groups showed altered levels compared to control groups that could be linked to doses and combined effects of each chemical substance (cypermethrin and piperonyl butoxide). Oxidative stress markers suggest that cypermethrin, piperonyl butoxide and the co-exposed groups, induce oxidative stress as well as induction of telomerase activity.

Obesity and thus, lipotoxicity, is a major health risk factor. Modern exposure to environmental chemicals has contributed significantly to the obesity epidemic. The purpose of this study was to assess, via telemedicine and using bioelectrical impedance analysis (BIA) in schools, the levels of adiposity and other body composition parameters of Greek adolescents in relation with their metabolic syndrome (MetS) characteristics. A representative sample (1575 adolescents, 14.4 ± 1.7 years-old) of the Attica region population, underwent body composition assessment of fat mass (FM), fat-free mass (FFM), and total body water (TBW) and was evaluated for anthropometric and MetS characteristics. Males demonstrated higher FFM% and TBW% but lower FM% than females. Adolescents with abdominal obesity/MetS (n = 149/n = 40) demonstrated significantly (P Findings suggest that early “osteosarcopenic” elements of abdominal obesity/MetS may exist even in adolescence. The application of BIA, incorporated in the new approach methodology of telemedicine in schools, identified adolescents at risk for obesity complications.

The occurrence of residues of DDT and its metabolites was monitored in 196 cow milk samples of various pasteurized commercial types collected from the Greek market. Residue levels were determined by GC-MS analysis. In 97.4% of the samples at least one DDT isomer or one of the DDT metabolites was detected, in levels not exceeding the maximum permitted residue level by the EU. Hazard Index for both carcinogenic and non-carcinogenic effects was estimated under two assumptions: a) using DDT concentrations from positive samples and b) imputing LOD/2 as an arbitrary concentration for negative samples. No statistically significant differences in detected or summed residue (p > 0.05) concentrations between different milk types were observed, with the exception of specific metabolites of DDT in some milk types. Exposure assessment scenarios were developed for children aged 1, 3, 5, 7 and 12 years old based on estimated body weights and daily milk consumption. Hazard Indices for non-carcinogenic effects were below 0.109 covering also carcinogenic effects according to WHO approach. The cancer risk values for carcinogenic effects according to the US EPA Cancer Benchmark Concentration approach, ranged from 0.4 to 18. For both effects the highest values were calculated for the 1- to 3-year-old age groups.

Lycopene, a carotenoid, is known for its antioxidant properties. Little is known, though, about the relationship of dietary tomato-juice intake and risks factors, like inflammation, insulin resistance and hyperlipidemia, implicated in metabolic syndrome. In the present study, we examined whether supplementation with tomato-juice has any implication on the risk status of patients with metabolic syndrome. A comparative study was conducted in 27 individuals diagnosed with metabolic syndrome. Fifteen of them were instructed to use commercially available tomato-juice as refreshment 4 times a week over a period of two months and twelve individuals served as the control group. Several parameters reflective of the metabolic syndrome were monitored both in the group supplemented with tomato juice and in the control group (ADMA for entdothelial function, TNF-α and IL-6 for inflammation, FIRI for insulin resistance). There was a significant improvement in the inflammation status and the endothelial dysfunction of the tomato-juice supplemented patients. At the same time, insulin resistance improved and a pronounced decrease in LDL was recorded, along with a slight increase in HDL. The results of the present study suggest an alleviating effect of tomato-juice with regard to risk factors associated with metabolic syndrome.

Creatinine phosphokinase (CPK) and lactate dehydrogenase (LDH) are important biological markers of various myocardial disorders and exercise-induced muscle damage. Lycopene, on the other side, is a natural anti-oxidant with protective action against cardiovascular risk. Fifteen anaerobically trained athletes with elevated LDH and CPK baseline levels were enrolled in this study after undergoing thorough biochemical and cardiovascular evaluation with echocardiocraphy. In nine athletes tomato juice, a lycopene plain juice, was administered during and after exercise, replacing the carbohydrate supplementation beverages commonly used during training for over a 2-month period. Tomato juice administration significantly reduced LDH and CPK levels, which returned back to almost normal levels. At the same time homocysteine and C-reactive protein were also attenuated. No changes were observed in the control group, where the usual carbohydrate supplementation had been followed.

Alzheimer's disease (AD) is the most common type of dementia accounting for 60-80% of the reported cases. The aim of this study was to evaluate levels of certain parameters of oxidative stress and markers of endothelial dysfunction in the blood of 21 AD patients under standard treatment compared with 10 controls, in an attempt to elucidate the contribution of AD to the total oxidative stress status of the patients. Results indicate that IL-6, TNF-α, ADMA and homocysteine levels were significantly elevated in AD patients. Protein carbonyls levels were higher in AD group, while glutathione reductase and total antioxidant capacity were lower, depicting decreased defense ability against reactive oxygen species. Besides, a higher level of advanced glycation end-products was observed in AD patients. Depending on the treatment received, a distinct inflammatory and oxidative stress profile was observed: in Rivastigmine-treated group, IL6 levels were 47% lower than the average value of the remaining AD patients; homocysteine and glutathione reductase were statistically unchanged in the Rivastigmine and Donepezil-Memantine, respectively Donepezil group. Although the study is based on a limited population, the results could constitute the basis for further studies regarding the effect of medication and diet on AD patients.

Purpose The aim of the present study was to investigate the effects of diazinon and propoxur on liver and kidneys, following long term exposure of rabbits. Methods Ten New Zealand white female rabbits were used. The animals were divided into 5 groups, consisting of 2 animals each. Diazinon (groups 1 and 2) and propoxur (groups 3 and 4) were administered at 2 different doses, and group 5 served as the control group. Histopathological lesions in the liver and kidneys, oxidative stress and oxidative DNA damage were evaluated. Results Both pesticides induced focal inflammation and fibrosis in the liver and kidneys. The low dose of propoxur induced a significant increase in total antioxidant capacity (TAC), with no difference in reduced glutathione (GSH), while the high dose of propoxur induced an increase in GSH with no change in TAC. For diazinon-exposed animals, the opposite findings were observed. Both diazinon and propoxur induced a statistically significant oxidative DNA damage in the liver and kidneys and a subsequent increase in telomerase activity in these tissues, possibly as a counteracting mechanism. Furthermore, systemic inflammation, as depicted by the dose-dependent increase in telomerase activity in peripheral blood mononuclear cells (PBMCs), was observed in propoxur treated animals. Conclusions Histopathological lesions, oxidative stress and genotoxic effects were induced in liver and kidneys following long term exposure of rabbits to diazinon and propoxur.

Exercise has been associated with oxidative stress that is correlated with muscle fatigue and reduced exercise performance. The aim of this study was to examine the effects of a special cake (consisting of carbohydrate to whey protein 3.5:1) vs an isocaloric carbohydrate cake on biomarkers of oxidative stress in 9 males after exhaustive cycling. A randomized single-blind cross-over study was completed. They performed one trial involving a 2-h exercise on a cycle ergometer at 60-65% VO(2)max followed by a 4-h recovery and then a second trial involved an 1-h exercise at 60-65% VO(2)max which was increased at 95% VO(2)max (time trial). The subjects received 4 experimental or placebo cakes after the first trial (the first immediately after and then one every hour). Blood samples were collected at eight time intervals: pre-exercise, 30 min, 1.5 h and 4 h post-exercise, post time Trial, 1 h, 24 h and 48 h post time Trial. Thiobarbituric Acid Reactive Substances (TBARS), protein carbonyls, total antioxidant capacity (TAC), catalase and glutathione (GSH) were determined spectrophotometrically. The mean time to exhaustion did not differ upon cake consumption. Consumption of the special cake reduced TBARS significantly, but had no effect on other oxidative stress markers.

The feeding behaviour of the pine processionary (PPC) caterpillar Thaumetopoea pityocampa Den. and Schiff. (Lepidoptera, Thaumetopoeidae) in L3-L4 stages was explored by means of laboratory arena feeding trials and natural substrates. In the bioassays, volatile extracts of 15 pine species, 8 of which are naturally growing in Greece, were incorporated. An analytical model was developed based on the principle of multinomial logit regression with five outcomes on the basis of the behavioural feeding sequence of the caterpillars. The outcomes were the five steps in which the feeding behavioural sequence was decomposed. The model's suitability (MacFadden's rho(2)=0.229, P

A selective gas-liquid chromatographic method with mass spectrometry (GC-MS) for the simultaneous confirmation and quantification of ephedrine, pseudo-ephedrine, nor-ephedrine, nor-pseudoephedrine, which are pairs of diastereoisomeric sympathomimetic amines, and methyl-ephedrine was developed for doping control analysis in urine samples. O-Trimethylsilylated and N-mono-trifluoroacetylated derivatives of ephedrines--one derivative was formed for each ephedrine--were prepared and analyzed by GC-MS, after alkaline extraction of urine and evaporation of the organic phase, using d3-ephedrine as internal standard. Calibration curves, with r2>0.98, ranged from 3.0 to 50 microg/ml depending on the analyte. Validation data (specificity, % RSD, accuracy, and recovery) are also presented.

An excretion study of the beta2-agonist 7-[3-[(beta-3,5-trihydroxyphenethyl)amino]-propyl]theophylline (reproterol) in human urine, which is reportedly misused by athletes and horses as a doping agent, is presented. The study was performed after an oral administration of 20 mg of reproterol hydrochloride. The collected urine samples were prepared using the standard anabolic steroid extraction procedure and analyzed by gas chromatography coupled with quadrupole mass spectrometry and, also, with high-resolution mass spectrometry (HRMS). The main reproterol metabolite was found, whereas unchanged reproterol was not detected. The structure of the main metabolite was confirmed by an accurate HRMS measurement of diagnostic ions. Finally, an excretion urine profile of the main metabolite is presented. The mass spectrum of another possible unidentified reproterol metabolite is also reported.

The first official report on the organization, analytical methodologies, and the results of the doping control analysis performed by the International Olympic Committee (IOC)-accredited Doping Control Laboratory of Athens, Greece, during the 6th World Championships of Athletics, held in Athens on 1–10 August 1997, is presented. The significance of the various analytical parameters of the doping control is shown. The results include 20 positive cases, of which two were detected in screening procedures and confirmed exclusively by high resolution mass spectrometry. The screening results and analytical data on the excretion of a new black-market doping agent, phenylpiracetam or carfedon, which was unknown in the literature, are also presented.

The regioselective acylation of monosaccharides, such as glucose, fructose, mannose and arabinose with lauric acid (C12) has been investigated in hexane using commercial Candida rugosa lipase (EC 3.1.1.3) and Candida antarctica (NOVO-ZYME) lipase immobilised on anionic resin. In order to overcome sugars insolubility in hexane, sugars were introduced in the reaction mixture preabsorbed on silica gel. The chemical structure of the products has been determined by 1 H NMR analysis. The two enzymes tested seemed to prefer hexoses rather than pentoses as the sugar substrate. They exhibited different water activity optimums as well as different optimum sugar/acyl donor molar ratios. Water activity control throughout the reaction and temperature increase from 30 to 50 °C enhanced sugar ester production using both lipases.

Oxidative stress has been shown to increase after acute myocardial infarction and during coronary reperfusion. Allopurinol inhibits xanthine oxidase, an enzyme involved in reperfusion injury. In this study, 40 patients with ST elevation myocardial infarction and symptoms' onset 3-12 h, who underwent primary coronary intervention, were administered either allopurinol (loading dose 400 mg followed by 100 mg for 1 month--group A, 21 patients), or placebo (group B). Allopurinol resulted in a more effective ST-E recovery (P