Olive oil with high polyphenolic content induces both beneficial and harmful alterations on rat redox status depending on the tissue.

Olive oil (OO) possesses a predominant role in the diet of Mediterranean countries. According to a health claim approved by the European Food Safety Authority, OO protects against oxidative stress­induced lipid peroxidation in human blood, when it contains at least 5 mg of hydroxytyrosol and its derivatives per 20 g. However, studies regarding the effects of a total OO biophenols on redox status in vivo are scarce and either observational and do not provide a holistic picture of their action in tissues. Following a series of in vitro screening tests an OO containing biophenols at 800 mg/kg of OO was administered for 14 days to male Wistar rats at a dose corresponding to 20 g OO/per day to humans. Our results showed that OO reinforced the antioxidant profile of blood, brain, muscle and small intestine, it induced oxidative stress in spleen, pancreas, liver and heart, whereas no distinct effects were observed in lung, colon and kidney. The seemingly negative effects of OO follow the recently formulated idea in toxicology, namely the real life exposure scenario. This study reports that OO, although considered a nutritional source rich in antioxidants, it exerts a tissues specific action when administered in vivo.

A mixture of routinely encountered xenobiotics induces both redox adaptations and perturbations in blood and tissues of rats after a long-term low-dose exposure regimen: The time and dose issue.

Exposure of humans to xenobiotic mixtures is a continuous state during their everyday routine. However, the majority of toxicological studies assess the in vivo effects of individual substances rather than mixtures. Therefore, our main objective was to evaluate the impact of the 12- and 18-month exposure of rats to a mixture containing 13 pesticides, food, and life-style additives in three dosage levels (i.e. 0.0025 × NOAEL, 0.01 × NOAEL, and 0.05 × NOAEL), on redox biomarkers in blood and tissues. Our results indicate that the exposure to the mixture induces physiological adaptations by enhancing the blood antioxidant mechanism (i.e., increased glutathione, catalase and total antioxidant capacity and decreased protein carbonyls and TBARS) at 12 months of exposure. On the contrary, exposure to the 0.05 × NOAEL dose for 18 months induces significant perturbations in blood and tissue redox profile (i.e., increased carbonyls and TBARS). This study simulates a scenario of real-life risk exposure to mixtures of xenobiotics through a long-term low-dose administration regimen in rats. The results obtained could support, at least in part, the necessity of introducing testing of combined stimuli at reference doses and long term for the evaluation of the risk from exposure to chemicals.

Genotoxic, cytotoxic, and cytopathological effects in rats exposed for 18 months to a mixture of 13 chemicals in doses below NOAEL levels.

The present study investigates the genotoxic and cytotoxic effects of long term exposure to low doses of a mixture consisting of methomyl, triadimefon, dimethoate, glyphosate, carbaryl, methyl parathion, aspartame, sodium benzoate, EDTA, ethylparaben, buthylparaben, bisphenol A and acacia gum in rats. Four groups of ten Sprangue Dawley rats (5 males and 5 females per group) were exposed for 18 months to the mixture in doses of 0xNOAEL, 0.0025xNOAEL, 0.01xNOAEL and 0.05xNOAEL (mg/kg bw/day). After 18 months of exposure, the rats were sacrificed and their organs were harvested. Micronuclei frequency was evaluated in bone marrow erythrocytes whereas the organs were cytopathologically examined by the touch preparation technique. The exposure to the mixture caused a genotoxic effect identified only in females. Cytopathological examination showed specific alterations of tissue organization in a tissue-type dependent manner. The observed effects were dose-dependent and correlated to various tissue parameters. Specifically, testes samples revealed degenerative and cellularity disorders, liver hepatocytes exhibited decreased glycogen deposition whereas degenerative changes were present in gastric cells. Lung tissue presented increased inflammatory cells infiltration and alveolar macrophages with enhanced phagocytic activity, whereas brain tissue exhibited changes in glial and astrocyte cells' numbers. In conclusion, exposure to very low doses of the tested mixture for 18 months induces genotoxic effects as well as monotonic cytotoxic effects in a tissue-dependent manner.

The European Registered Toxicologist (ERT): Current status and prospects for advancement.

Following its inception in 1994, the certification of European Registered Toxicologists (ERT) by EUROTOX has been recognized as ensuring professional competence as well as scientific integrity and credibility. Criteria and procedures for registration are contained in the ERT "Guidelines for Registration 2012". The register of ERT currently has over 1900 members. In order to continue the harmonisation of requirements and processes between national registering bodies as a prerequisite for official recognition of the ERT title as a standard, and to take account of recent developments in toxicology, an update of the ERT Guidelines has been prepared in a series of workshops by the EUROTOX subcommittees for education and registration, in consultation with representatives of national toxicology societies and registers. The update includes details of topics and learning outcomes for theoretical training, and how these can be assessed. The importance of continuing professional development as the cornerstone of re-registration is emphasised. To help with the process of harmonisation, it is necessary to collate and share best practices of registration conditions and procedures across Europe. Importantly, this information can also be used to audit compliance with the EUROTOX standards. As recognition of professionals in toxicology, including specialist qualifications, is becoming more important than ever, we believe that this can best be achieved based on the steps for harmonisation outlined here together with the proposed new Guidelines.

Citizen scientist lepidopterists exposed to potential carcinogens.

Lepidopterists use substantial volumes of solvents, such as chloroform, 1,1,2,2-tetrachloroethane and xylene, in their traps when collecting faunistic and phenological data. A majority of them are citizen scientists and thus in part not identified by occupational healthcare as being at risk due to solvent handling. We surveyed the extent of solvent use, the frequency and extent of potential exposure and the safety precautions taken in trapping and catch handling by Finnish lepidopterists. Chloroform and 1,1,2,2-tetrachloroethane were the most frequently used anaesthetics. Potential for exposure prevailed during trap maintenance and exploration and catch sorting. Adequate protection against vapours or spills was worn by 17% during trap exploration. Subjects completed a median of 100 trap explorations per season. Dermal or mucosal spills were recorded at a median rate of one spill per ten (chloroform) to 20 (1,1,2,2-tetrachloroethane and xylene) trap explorations. Median annual cumulative durations of 8 and 20 h of exposure to chloroform and 1,1,2,2-tetrachloroethane at levels above odour detection threshold were reported. Subjective adverse findings possibly related solvents had been noticed by 24 (9.8%) lepidopterists. All the events had been mild to moderate. No factor predicting unsafe procedures or adverse reactions was recorded despite thorough statistical testing.

Multicomponent analysis of replacement liquids of electronic cigarettes using chromatographic techniques.

The electronic cigarette (e-cig) is an invention of the past few years and its popularity is rapidly growing all over the world. A rapid multicomponent analytical protocol for the analysis of the replacement liquids (e-liquids) of e-cig was developed using gas (GC) and liquid chromatography (LC)-mass spectrometry (MS). GC-MS-based methods were developed for the determination of the main humectants and polycyclic aromatic hydrocarbons (PAHs). For the determination and quantification of nicotine (NIC) and nitrosamines, appropriate LC-MS-based methods were developed. The approbated methods were applied for the analysis of 263 e-liquid samples obtained from the Greek market. The instruments response was linear; the limits of quantification ranged from 0.003 µg/mL for three PAHs to 1.187 µg/mL for glycerol. The precision was <16% for all analytes, while the mean accuracy ranged from 99.1% for NIC to 106.6% for the flavor 2,5-dimethylpyrazine. The measured concentrations of NIC were correlated with the theoretical concentrations as reported by the manufacturers. An analog relation between the concentration of the glycerol and of propylene glycol was noticed. The frequency of detection of flavors ranged from 30.4% for the methyl cyclopentenolone to 5.3% for 3.4-dimethoxybenzaldehyde. Nitrosamines and PAHs were not detected in any sample. Because a similar analytical protocol was not available from the existing literature so far, our study offers the advantage of complete analytical methods for rapid and simultaneous multicomponent identification.

Determination of buprenorphine, norbuprenorphine and naloxone in fingernail clippings and urine of patients under opioid substitution therapy.

The aim of this study was to develop and validate a method for the determination of buprenorphine (BUP), norbuprenorphine (NBUP) and naloxone (NAL) in fingernails and urine samples collected from former heroin users under suboxone substitution therapy. The analytes were extracted by solid-liquid or solid-phase extraction and were analyzed by liquid chromatography-mass spectrometry. The validation of the analytical methods developed included linearity, recovery, accuracy, precision, ion suppression, sensitivity of interfaces and limits of determination and quantification. The validated methods were applied to samples from 46 individuals. The majority of the urine samples were positive for all analytes (93.5% for BUP, 95.7% for NBUP and 84.8% for NAL). In nails, a higher detection rate was observed for NBUP and BUP (89.1%), compared with NAL (10.9%). The median values of the NBUP/BUP and the NAL/BUP ratio were 2.5 and 0.3 in urine and 0.8 and 0.3 in nails, respectively. A statistically significant correlation was found between the BUP, NBUP and total BUP (BUP and NBUP) concentrations in urine and those in nails. A weak correlation was observed between the daily dose (mg/day) and total BUP (P = 0.069), or NBUP (P = 0.072) concentrations in urine. In contrast, a strong correlation was found between the total amount of BUP administered during the last 12 months and total BUP (P = 0.038), or NBUP (P = 0.023) concentrations in urine. Moreover urine BUP, NBUP and total BUP concentrations correlated significantly. Our study demonstrated successfully the application of the developed method for the determination of the three analytes in urine and nails.

Administration of tomato juice ameliorates lactate dehydrogenase and creatinine kinase responses to anaerobic training.

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.

Pesticides and oncogenic modulation.

Pesticides constitute a diverse class of chemicals used for the protection of agricultural products. Several lines of evidence demonstrate that organochlorine and organophosphate pesticides can cause malignant transformation of cells in in vitro and in vivo models. In the current minireview a comprehensive summary of recent in vitro findings is presented along with data reported from human population studies, regarding the impact of pesticide exposure on activation or dysregulation of oncogenes and tumor suppressor genes. Substantial mechanistic work suggests that pesticides are capable of inducing mutations in oncogenes and increase their transcriptional expression in vitro, whereas human population studies indicate associations between pesticide exposure levels and mutation occurrence in cancer-related genes. Further work is required to fully explore the exact mechanisms by which pesticide exposure affects the integrity and normal function of oncogenes and tumor suppressor genes in human populations.

A mechanistic overview of health associated effects of low levels of organochlorine and organophosphorous pesticides.

Organochlorine and organophosphate pesticides are compounds that can be detected in human populations as a result of occupational or residential exposure. Despite their occurrence in considerably low levels in humans, their biological effects are hazardous since they interact with a plethora of enzymes, proteins, receptors and transcription factors. In this review we summarize the cell and molecular effects of organochlorine and organophosphate pesticides with respect to their toxicity, with particular emphasis on glucose and lipid metabolism, their interaction with some members of the nuclear receptor family of ligand-activated transcription factors, including the steroid and peroxisome proliferator activated receptors that changes the expression of genes involved in lipid metabolism and xenobiotic detoxification. More importantly, evidence regarding the metabolic degradation of pesticides and their accumulation in tissues is presented. Potential non-cholinergic mechanisms after long-term low-dose organophosphate exposure resulting in neurodevelopmental outcomes and neurodegeneration are also addressed. We conclude that the mechanism of pesticide-mediated toxicity is a combination of various enzyme-inhibitory, metabolic and transcriptional events acting at the cellular and molecular level.

Hair as a biomarker of polybrominated diethyl ethers' exposure in infants, children and adults.

Over the last 20 years hair has moved from being a highly questionable biological matrix to mainstream and acceptable biomarker in forensic sciences where it is primarily used to determine past and present exposure to illicit drugs. In contrast, the use of hair to assess exposure to pesticides and persistent environmental pollutants is still not common. The applicability of this matrix to assess an individual's body burden of chemicals such as polybrominated diethyl ethers (PBDEs) can provide critical insight into current, but also to past exposure levels, which is not possible with more conventional matrices such as blood and urine. Furthermore, as PBDEs cross the placenta and since the hair the fetus is born with begins to grow during the third trimester, this matrix can be used to assess in utero exposure. These features of hair may therefore be used to determine the potential roles of chemicals such as PBDEs in mediating physiological or anatomical abnormalities in infants, children or adults.

Monitoring of systemic exposure to plant protection products and DNA damage in orchard workers.

The systemic exposure of plum tree growers and operators to plant protection products (PPPs) and effects on DNA were assessed. Specifically, a GC-MS/MS method was developed and validated for the analysis of serum samples for the presence of seven active substances of PPPs. The analytical results verified the presence of myclobutanil, propargite, cypermethrin and deltamethrin in 7 out of 19 serum samples. The incidence of DNA damage was monitored using the single cell electrophoresis assay (comet assay). A paired Student's t-test revealed a statistically significant increase of SSBs in the blood samples collected at the end of the cropping period as compared to the samples collected from the same subjects before the start of PPPs application period. Moreover, the group of seven subjects with detectable serum pesticides levels revealed statistically significant increase of SSBs as compared to the group of subjects with no detectable PPP levels. The results of the present study demonstrate that the agriculture workers may exhibit detectable level of systemic exposure to the applied PPPs which are correlated to increased DNA damage during the cultivation period.

Workplace health promotion activities of Finnish occupational health nurses.

OBJECTIVE: This study describes workplace centered activities performed by occupational health professionals, as well as difficulties linked to workplace centered activities, cooperation with client workplaces, and participation both in risk assessment and in the development of working environment. DESIGN: The design is a cross-sectional prevalence study. SAMPLE: The questionnaires were sent to 250 occupational health professionals, of whom 176 (70%) returned the completed forms and of whom 99% were nurses. MEASUREMENTS: Their activities, difficulties, cooperation, and participation in risk assessment and development of safe and healthy working conditions. ANALYTIC STRATEGY: The data were analyzed by using frequencies, means, 1-way ANOVA, and the Kruskal-Wallis test. RESULTS: Workplace focused activity hours were mainly spent on risk assessment and counseling. One workplace visit took approximately 4 hr. The greatest difficulties were encountered with participation in development projects and in monitoring the effectiveness of improvement activities, especially in health care centers. Occupational health nurses cooperated with client enterprises and participated in risk assessment more often than the other occupational health professionals. Also workplace representatives participated actively in risk assessment. The most difficult modules were considered to be the risk assessment of chemicals, biological factors, and perceived stress, this being especially so in health care centers. CONCLUSIONS: It is essential that occupational health professionals visit more often in their client enterprises and improve their skills in preventing harms caused by chemical and biological factors, and perceived stress.

Analytical investigation: assay of D-lactate in diabetic plasma and urine.

OBJECTIVE: In this study we developed a 96-micro plate enzymatic assay for D-lactate in plasma and urine. METHODS: D-lactate was assayed enzymatically with a UV-spectrophotometer in plasma from 38 and in urine from 37 diabetics and from 42 non-diabetic controls. RESULTS: The within-run coefficients of variations (CV) were 2.6% for plasma and 5.7% for urine. The between run CVs were 6.8% for plasma and 6.7% for urine. The mean recovery with standard deviation (S.D.) was 107.4+/-7.3% for plasma and 100.1%+/-6.7% for urine. The plasma D-lactate in diabetics were (mean+/-S.D.) 39.6+/-23.7 microM. We found significant difference between the urinary d-lactate in controls and diabetics (18.2+/-12.0 vs. 35.9+/-24.2 microM/mM creatinine, p<0.0001). CONCLUSIONS: The assay proved reliable with acceptable precision and recovery. Results suggest that diabetics have elevated urinary and plasma D-lactate as compared to controls.

Occupational risk identification using hand-held or laptop computers.

This paper describes the Work Environment Profile (WEP) program and its use in risk identification by computer. It is installed into a hand-held computer or a laptop to be used in risk identification during work site visits. A 5-category system is used to describe the identified risks in 7 groups, i.e., accidents, biological and physical hazards, ergonomic and psychosocial load, chemicals, and information technology hazards. Each group contains several qualifying factors. These 5 categories are colour-coded at this stage to aid with visualization. Risk identification produces visual summary images the interpretation of which is facilitated by colours. The WEP program is a tool for risk assessment which is easy to learn and to use both by experts and nonprofessionals. It is especially well adapted to be used both in small and in larger enterprises. Considerable time is saved as no paper notes are needed.

Changes in serum markers indicative of health effects in vineyard workers following exposure to the fungicide mancozeb: an Italian study.

The aim of this study was to investigate the health effects induced by exposure to the fungicide mancozeb in Italian vineyard workers. Ninety-three Italian subjects entered the study - 48 vine-growers intermittently exposed to mancozeb and 45 healthy controls. The subjects were investigated three times: before the seasonal application of pesticides (T0), 30 days after the beginning of the application period (T30), and 45 days after T0 (T45). At T0 the comparison between agricultural workers and controls showed a higher prevalence of cold or flu symptoms, a statistically significant lower percentage of monocytes, higher absolute count of T lymphocytes, CD4 and natural killer cells, and lower plasma levels of IgA and IgM in workers. Such differences were not confirmed at T30 and T45. In fact at T30 in exposed workers, besides a significant increase of urinary ethylenethiourea, confirming mancozeb exposure, T lymphocytes, CD4 and natural killer cells, IgA and IgM returned to values comparable to those observed in controls. Moreover, no other differences in clinical signs, haematological, and immune parameters, such as the immune functional capability evaluated as a response to hepatitis B vaccination, was observed. Altogether the differences between exposed and controls were not consistently correlated to any clinical impairment and suggest that the seasonal application of mancozeb does not pose a significant health risk to exposed subjects.

Work activity analysis of Finnish occupational health professionals.

BACKGROUND: Some Finnish studies have dealt with how occupational health nurses divide their working hours but other occupational health professionals have not been evaluated. AIMS: This study describes how occupational health professionals allocate their working hours between main tasks. METHODS: Questionnaires were sent to 250 occupational health professionals, of whom 176 (70%) returned the completed forms. The data were analysed by using frequencies, means and one-way analysis of variance test. RESULTS: Employee-oriented tasks accounted for roughly 50% of working hours from all occupational health professionals. The remaining working hours were shared between workplace visits, co-operation with partners, other occupational health care responsibilities and tasks in other health care fields, especially in the health care centres. These working hours varied greatly between the different occupational health professional groups. All units employed full-time occupational health nurses, but the services of physicians, physiotherapists and psychologists were usually provided part-time or even restricted to a few hours each week because these services were difficult to obtain. Occupational health nurses working in the municipal health care centres spent more time on workplace visits than other nurses. Employee-oriented tasks were emphasized more in physicians', physiotherapists' and psychologists' work, especially in private medical health care units and in the jointly owned health care units. CONCLUSIONS: The amount of time occupational health professionals are able to spend on workplace activities appears to be determined by the type of their employer.

History and results of the two inter-laboratory round robin endotoxin assay studies on cotton dust.

BACKGROUND: In the US cotton industry, airborne cotton dust levels are regulated, and other countries are moving to specify safety limits for airborne endotoxins. There is concern about potential respiratory health hazards associated with agricultural and other organic dusts. In laboratories, ranking which samples have high and low levels of endotoxin is usually in good agreement between laboratories. When different laboratories assay identical samples, the levels differ. The objective of this research was to evaluate the intra- and inter-laboratory variability for 13 laboratories measuring endotoxin in cotton dust. METHOD: Two inter-laboratory round robin endotoxin assay studies were conducted using cotton dust. In the first round robin, each laboratory used their normal in-house assay method and then used a common extraction protocol. In the second round robin, a common extraction protocol and endotoxin assay kit was used. RESULTS: The intra-laboratory results had small variations but inter-laboratory results had very high variations. The inter-laboratory results using a common extraction protocol showed reduced differences. Using the same extraction protocol and endotoxin assay kit, the intra-laboratory variation was small and inter-laboratory variation was reduced but not enough for inter-laboratory agreement. Most of the laboratories were able to discern between the high and low endotoxin concentration dusts. CONCLUSIONS: Standardization has reduced the differences in results between laboratories and possibly further standardization may bring closer inter-laboratory agreement.