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.

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.

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.

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.

Evaluation of exposure to 1-alkoxy-2-propanols and 1-(2-methoxy-1-methylethoxy)-2-propanol by the analysis of the parent compounds in urine.

Floor lacquerers' inhalation and total exposure to 1-alkoxy-2-propanols and 1-(2-methoxy-1-methylethoxy)-2-propanol (DPGME) were measured. The total exposure was biomonitored by urinalysis of free unchanged 1-alkoxy-2-propanols and DPGME. The floor lacquerers' 8-h inhalation exposures to 1-methoxy-2-propanol (PGME), 1-butoxy-2-propanol (PGBE) and DPGME were 1.9+/-1.3 (mean+/-S.D., n=15), 1.0+/-1.4ppm (n=11) and 0.2+/-0.3ppm (n=11), respectively. The gravity-corrected urinary excretions of PGME, PGBE and DPGME were 5.3+/-5.4mumol/l, 0.9+/-0.9mumol/l and 1.5+/-2.8mumol/l, respectively. A linear relationship was found between the gravity-corrected urinary excretion of PGME (R(2)=0.82), PGBE (R(2)=0.93) and DPGME (R(2)=0.93) and their preceding 8-h inhalation exposure. The correlations between the uncorrected urinary excretions and inhalation exposures to PGME, PGBE and DPGME was also calculated and found good (R(2)=0.82-0.95). The effect of work strain on the total exposure seemed to be more relevant in the exposure to hydrophilic PGME than in the exposure to more lipophilic PGBE.

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.

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.

Serotonergic modulation of mismatch negativity.

Neurochemical mechanisms mediating the interaction between emotional and cognitive processing are not yet fully understood. Here, we utilized acute tryptophan depletion (ATD) to reduce the brain synthesis of serotonin (5-HT), which is thought to have a central role in regulation of emotions and mood in humans. ATD effects on event-related potentials and magnetic fields were studied using a passive odd-ball paradigm in a randomized, double-blinded, controlled, cross-over design. Auditory responses were recorded simultaneously with high-resolution magnetoencephalography (MEG) and electroencephalography (EEG) in 14 healthy subjects, 5 h after ATD or a control condition. ATD significantly increased depressed mood and lowered plasma tryptophan concentration (total tryptophan decreased by 75%, free tryptophan decreased by 39%). As compared with the control condition, ATD increased the amplitudes of mismatch negativity (MMN) to duration and frequency changes and decreased the latencies of magnetic MMN to frequency changes in the hemisphere ipsilateral to the ear stimulated. Further, ATD modulated N1m latencies and decreased P2m source activity. ATD increased the interhemispheric latency difference of MMNm to frequency changes. No effects on P50 were observed. The present results suggest serotonergic modulation of preattentive auditory change detection, suggested to initiate involuntary attention shifting in the human brain.

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.

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.

Serotonin modulates early cortical auditory processing in healthy subjects: evidence from MEG with acute tryptophan depletion.

We studied the effects of acute tryptophan depletion (ATD) on early cortical auditory processing. Middle-latency auditory evoked fields (MAEF) were investigated in 14 healthy subjects after 5 h of ATD or control mixture ingestion in a randomized, double-blinded, controlled cross-over design study. MAEFs to monaural click stimuli (0.1-ms duration) were recorded with a 122-channel neuromagnetometer. Total plasma tryptophan (Trp), free Trp, and large neutral amino acid (LNAA) concentrations were determined by using high-performance liquid chromatography. ATD lowered the total plasma Trp levels by 75%, free Trp level by 47%, and the ratio Trp/SigmaLNAA by 92%. The control mixture increased total Trp level by 45% and free Trp by 32%, and decreased the ratio Trp/SigmaLNAA by 35%. The ratio tyrosine/SigmaLNAA did not differ between ATD and control experiment. ATD resulted in a significant main effect on Pam latencies and a near-significant main effect on Pam amplitudes. A significant Mixture ingestion X Sex interaction on Nbm amplitude and a significant Mixture ingestion X Sex X Hemisphere interaction on Pam latency were observed. ATD did not affect the MAEF source dipoles. The Pam latencies in both hemispheres had a significant negative relationship with the extent of ATD. The results suggest that the neurotransmitter serotonin is involved in early auditory cortical processing. Further, the serotonin modulation may be different in males and females.

Acute tryptophan depletion decreases intensity dependence of auditory evoked magnetic N1/P2 dipole source activity.

RATIONALE: Intensity dependence of the N1/P2 components may be regulated by serotonergic neurons in the primary auditory cortex, where low activity leads to a high intensity dependence and vice versa. Depletion of tryptophan (TRP), a precursor for serotonin has been described to reduce serotonin content in brain of animals and humans. OBJECTIVE: We investigated the intensity dependence of magnetic and electric N1/P2 components in ten subjects in a double-blind, controlled, cross-over design study after oral mixture of amino-acids leading to acute tryptophan depletion (ATD) and control. METHODS: Auditory evoked magnetic fields (AEF) and potentials (AEP) were recorded with 122-channel magnetoencephalography simultaneously with 64-channel EEG 5 h after ingestion of mixtures. The AEF sources and strength were estimated by a least-squares fit of a single equivalent current dipole. The amplitudes and latencies of N1 and P2 recorded with EEG were analyzed at frontal electrode site. RESULTS: TRP depletion decreased the total and free TRP levels by 76 and 45% and control mixture increased it by 48 and 28%. ANOVA showed that ATD had a significant main effect on the N1m/P2m dipole moments at the contralateral ( P=0.02), but failed significantly to influence the ipsilateral responses. A significant mixture ingestion-by-stimulus intensity interaction was observed on the N1m/P2m dipole moments at the contralateral hemisphere ( P=0.01). The N1/P2 slope for intensity dependence function was decreased following ATD compared with the control experiment ( P=0.01) at the contralateral hemisphere. For EEG, a significant mixture ingestion-by-stimulus intensity interaction on the N1 latencies at the Fz electrode position was observed ( P=0.01). CONCLUSION: ATD decreased the intensity dependence of N1m/P2m source dipole moments in the primary auditory cortex at the hemisphere contralateral to the ear stimulated. These results suggest that serotonin participates in the regulation of intensity of auditory stimulation.

Acute tryptophan depletion does not change somatosensory evoked magnetic fields.

RATIONALE: Serotonin is shown to regulate the activity of primary auditory cortex, but little is known about serotonin modulation of other sensory cortices. METHODS: We investigated somatosensory evoked magnetic fields (SEF) to left median nerve stimulation in eight healthy subjects in a double-blind, controlled, cross-over design study after acute tryptophan depletion (ATD) and control mixture. SEFs were recorded with the whole-head magnetoencephalography 6 h after ingestion of mixtures. The SEF sources and strength were estimated by a least-squares fit of a single equivalent current dipole. RESULTS: ATD decreased the total and free TPR levels by 75 and 48% and control mixture increased them by 98% and 44%. ATD had no effect on the amplitudes or latencies of SEF components. The source locations of the responses were not significantly affected by ATD. CONCLUSION: Serotonin does not affect stimuli processing in the primary somatosensory cortex.

Auditory selective attention modulated by tryptophan depletion in humans.

To elucidate serotonin modulation of selective attention, 13 volunteers (21-30 years) were studied in two sessions, 5 h after either acute tryptophan depletion (ATD) that decreases brain serotonin synthesis, or control-mixture ingestion (randomized, double-blind, cross-over design). Simultaneous electroencephalogram and magnetoencephalogram were measured during dichotic listening of two concurrent trains of standard and deviant tones. Subjects counted the deviants presented to one ear and ignored those presented to the other ear. ATD lowered plasma total tryptophan by 75% and free tryptophan by 39%. ATD suppressed the amplitude enhancement of P50 and N1 to selectively attended tones, but did not affect the later aspects of processing negativity. The P50 latencies were increased after ATD, irrespective of attention. In conclusion, serotonin may regulate attentional modulation of early cortical stimulus processing.

Time to pregnancy among the wives of Finnish greenhouse workers.

OBJECTIVES: This study investigated the possibility of men's work in greenhouses and their exposure to pesticides being associated with reduced fertility. METHODS: A study on time to pregnancy was conducted among the families of Finnish male greenhouse employers and employees. Exposure to pesticides was assessed on the basis of questionnaire information and data gathered from the enterprises. Fecundability density ratios (FDR) for occupational exposure were calculated with discrete proportional hazards regression analyses. RESULTS: After three mailings, 578 (43%) couples participated. Fecundability was suggestively decreased for exposed greenhouse workers who were inefficiently protected, with FDR values of 0.67 [95% confidence interval (95% CI) 0.33-1.35], 0.92 [95% CI 0.45-1.88] and 0.77 [95% CI 0.46-1.29] for high exposure, moderate exposure and low exposure, respectively, as compared with unexposed greenhouse workers. The exposed men who efficiently used personal protective equipment were as fertile as the unexposed greenhouse workers. Exposure to pyrethroids (FDR 0.40, 95% CI 0.19-0.85) was related to decreased fecundability. Suggestive associations were observed for organophosphates (FDR 0.70, 95% CI 0.42-1.17) and carbamates (FDR 0.55, 95% CI 0.27-1.11). CONCLUSIONS: The findings of the study provide limited support for the hypothesis that exposure to pesticides is associated with reduced fertility. The findings for pyrethroids, organophosphates, and carbamates can serve as a basis for a hypothesis for future studies.

The efficacy of benzyl benzoate sprays in killing the storage mite Tyrophagus putrescentiae (Acari: Acaridae).

This study tested the effectiveness of benzyl benzoate (BB) in killing Tyrophagus putrescentiae (Schrank) mites when using the method utilized by regular consumers. The efficacy of the BB sprays was determined in laboratory experiments and semi-field experiments with mattress and mattress pad pieces. The mites were counted and their living status determined at different time points microscopically. In the laboratory experiment, the sprays containing either 0.5 %/0.9 % BB with 70 % ethanol or 0.1 % BB with absolute ethanol were highly efficient, resulting in over 90 % mite mortality within 20-30 minutes. In the semi-field experiment, mimicking the home application, the sprays were applied to pieces of a mattress and a mattress pad, and allowed to affect the area for 30 minutes before thorough vacuuming. The recovery of mites was usually less than 10 %. The sprays containing BB were effective in killing the mites in the laboratory, but success was less prominent in the semi-field tests. This method could be used in testing other compounds for their efficacy in killing mites.

Second inter-laboratory study comparing endotoxin assay results from cotton dust.

Previously, a large two-part inter-laboratory round robin endotoxin assay study was completed. This first study showed that when cotton dust samples, which are practically identical, are assayed for endotoxin that the intra- laboratory results had a very small variation while intra-laboratory results of the sample had a very high variation. In the first part of the study, each laboratory followed its own in-house assay protocol; but in the second part of the study, when the extraction protocol was standardized, the inter-laboratory results showed a lower variation, which suggested that with further standardization, further reduction of differences between laboratories might be achieved in order that results between laboratories would become more comparable. The results stimulated interest in extending the study to include cotton dust with two levels of endotoxin, standardization of the extraction protocol, and using the same assay kit from the same production lot. The results of this second round robin endotoxin assay study indicate that differences between laboratories are still high, but most of the laboratories could discern the cotton dusts with the different levels of endotoxin.

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.