Genotoxicity biomarkers in occupational exposure to formaldehyde: the case of histopathology laboratories

Formaldehyde, classified by the IARC as carcinogenic in humans and experimental animals, is a chemical agent that is widely used in histopathology laboratories. The exposure to this substance is epidemiologically linked to cancer and to nuclear changes detected by the cytokinesis-block micronucleus test (CBMN). This method is extensively used in molecular epidemiology, since it provides information on several biomarkers of genotoxicity, such as micronuclei (MN), which are biomarkers of chromosomes breakage or loss, nucleoplasmic bridges (NPB), common biomarkers of chromosome rearrangement, poor repair and/or telomere fusion, and nuclear buds (NBUD), biomarkers of elimination of amplified DNA. The aim of this study is to compare the frequency of genotoxicity biomarkers, provided by the CBMN assay in peripheral lymphocytes and the MN test in buccal cells, between individuals occupationally exposed and non-exposed to formaldehyde and other environmental factors, namely tobacco and alcohol consumption. The sample comprised two groups: 56 individuals occupationally exposed to formaldehyde (cases) and 85 unexposed individuals (controls), from whom both peripheral blood and exfoliated epithelial cells of the oral mucosa were collected in order to measure the genetic endpoints proposed in this study. The mean level of TWA8h was 0.16±0.11ppm (

Genotoxicity biomarkers in occupational to formaldehyde in pathology anatomy laboratories

Formaldehyde (FA) the most simple and reactive of all aldehydes, is a colorless, reactive and readily polymerizing gas at normal temperature. It has a pungent, suffocating odour that is recognized by most human subjects at concentrations below 1ppm. According to the Report on Carcinogens, FA ranks 25th in the overall U.S. chemical production with more than 11 billion pounds (5 million tons) produced each year. Is an important industrial compound that is used in the manufacture of synthetic resins and chemical compounds such as lubricants and adhesives. It has also applications as a disinfectant, preservative and is used in cosmetics. Estimates of the number of persons who are occupationally exposed to FA indicate that, at least at low levels, may occur in a wide variety of industries. The occupational settings with most extensive use of formaldehyde is in the production of resins and in anatomy and pathology laboratories. Several studies reported a carcinogenic effect in humans after inhalation of FA, in particular an increased risk for nasopharyngeal cancer. Nowadays, the International Agency for Research on Cancer (IARC) classifies FA as carcinogenic to humans (group 1), on the basis of sufficient evidence in humans and sufficient evidence in experimental animals. Manifold in vitro studies clearly indicated that FA is genotoxic. FA induced various genotoxic effects in proliferatin cultured mammalian cells. A variety of evidence suggests that the primary DNA alterations after FA exposure are DNA-protein crosslinks. Incomplete repair of DPX can lead to the formation of mutations.

Risk assessment in occupational exposure to formaldehyde: differences between anatomy and pathology laboratories and formaldehyde-resins production

Formaldehyde (CH2O), the most simple and reactive of all aldehydes, is colorless, and readily polymerizing gas at normal temperature. The most extensive use is in production of resins and has an important application as a disinfectant and preservative, reason why relevant workplace exposure may also occur in pathology and anatomy laboratories and in mortuaries. A study was carried out in Portugal, in a formaldehyde production resins factory and in 10 pathology and anatomy laboratories. It was applied a risk assessment methodology based on Queensland University proposal that permitted to perform risk assessment for each activity developed in a work station. This methodology was applied in 83 different activities developed in the laboratories and in 18 activities of the factory. Also, Micronucleus Test was performed in lymphocytes from 30 factory workers and 50 laboratories workers.

Occupational exposure to formaldehyde in anatomy and pathology laboratories: differences between exposure groups?

Formaldehyde, also known as formalin, formal and methyl aldehydes, is a colorless, flammable, strong-smelling gas. It has an important application in embalming tissues and that result in exposures for workers in the pathology anatomy laboratories and mortuaries. To perform exposure assessment is necessary define exposure groups and in this occupational setting the technicians and pathologists are the most important groups. In the case of formaldehyde, it seems that health effects are more related with peak exposures than with exposure duration.

Genotoxicity biomarkers: application in histopathology laboratories

Most cancers results from man-made and natural environmental exposures (such as tobacco smoke; chemical pollutants in air, water, food, drugs; radon; and infectious agents) acting in concert with both genetic and acquired characteristics. It has been estimated that without these environmental factors, cancer incidence would be dramatically reduced, by as much as 80%-90%. The modulation of environmental factors by host susceptibility was rarely evaluated. However, within the past few years, the interaction between environmental factors and host susceptibility factors has become a very active area of research. Molecular biology as a tool for use in epidemiological studies has significant potential in strengthening the identification of cancers associated with environmental exposures related to lifestyle, occupation, or ambient pollution. In molecular epidemiology, laboratory methods are employed to document the molecular basis and preclinical effects of environmental carcinogenesis. Molecular epidemiology has become a major field of research and considerable progress has been made in validation and application of biomarkers and its greatest contribution has been the insights provided into interindividual variation in human cancer risk and the complex interactions between environmental factors and host susceptibility factors, both inherited and acquired, in the multistage process of carcinogenesis.

Genotoxic effects in occupational exposure to formaldehyde: A study in anatomy and pathology laboratories and formaldehyde-resins production

Background - According to the Report on Carcinogens, formaldehyde ranks 25th in the overall U.S. chemical production, with more than 5 million tons produced each year. Given its economic importance and widespread use, many people are exposed to formaldehyde environmentally and/or occupationally. Presently, the International Agency for Research on Cancer classifies formaldehyde as carcinogenic to humans (Group 1), based on sufficient evidence in humans and in experimental animals. Manyfold in vitro studies clearly indicated that formaldehyde can induce genotoxic effects in proliferating cultured mammalian cells. Furthermore, some in vivo studies have found changes in epithelial cells and in peripheral blood lymphocytes related to formaldehyde exposure. Methods - A study was carried out in Portugal, using 80 workers occupationally exposed to formaldehyde vapours: 30 workers from formaldehyde and formaldehyde-based resins production factory and 50 from 10 pathology and anatomy laboratories. A control group of 85 non-exposed subjects was considered. Exposure assessment was performed by applying simultaneously two techniques of air monitoring: NIOSH Method 2541 and Photo Ionization Detection equipment with simultaneously video recording. Evaluation of genotoxic effects was performed by application of micronucleus test in exfoliated epithelial cells from buccal mucosa and peripheral blood lymphocytes. Results - Time-weighted average concentrations not exceeded the reference value (0.75 ppm) in the two occupational settings studied. Ceiling concentrations, on the other hand, were higher than reference value (0.3 ppm) in both. The frequency of micronucleus in peripheral blood lymphocytes and in epithelial cells was significantly higher in both exposed groups than in the control group (p < 0.001). Moreover, the frequency of micronucleus in peripheral blood lymphocytes was significantly higher in the laboratories group than in the factory workers (p < 0.05). A moderate positive correlation was found between duration of occupational exposure to formaldehyde (years of exposure) and micronucleus frequency in peripheral blood lymphocytes (r = 0.401; p < 0.001) and in epithelial cells (r = 0.209; p < 0.01). Conclusions - The population studied is exposed to high peak concentrations of formaldehyde with a long-term exposure. These two aspects, cumulatively, can be the cause of the observed genotoxic endpoint effects. The association of these cytogenetic effects with formaldehyde exposure gives important information to risk assessment process and may also be used to assess health risks for exposed workers.

Supply chain optimization of residual forestry biomass for bioenergy production: the case study of Portugal

Within a large set of renewable energies being explored to tackle energy sourcing problems, bioenergy can represent an attractive solution if effectively managed. The supply chain design supported by mathematical programming can be used as a decision support tool to the successful bioenergy production systems establishment. This strategic decision problem is addressed in this paper where we intent to study the design of the residual forestry biomass to bioelectricity production in the Portuguese context. In order to contribute to attain better solutions a mixed integer linear programming (MILP) model is developed and applied in order to optimize the design and planning of the bioenergy supply chain. While minimizing the total supply chain cost the production energy facilities capacity and location are defined. The model also includes the optimal selection of biomass amounts and sources, the transportation modes selection, and links that must be established for biomass transportation and products delivers to markets. Results illustrate the positive contribution of the mathematical programming approach to achieve viable economic solutions. Sensitivity analysis on the most uncertain parameters was performed: biomass availability, transportation costs, fixed operating costs and investment costs. (C) 2015 Elsevier Ltd. All rights reserved.

XRCC3 THR241MET polymorphism influence on nuclear buds frequency in exposed to formaldehyde

Formaldehyde (FA), also known as formalin, formal and methyl aldehydes, is a colorless, flammable, strong-smelling gas. It has an important application in embalming tissues and that result in exposures for workers in the pathology anatomy laboratories and mortuaries. Occupational exposure to FA has been shown to induce nasopharyngeal cancer and has been classified as carcinogenic to humans (group 1) on the basis of sufficient evidence in humans and sufficient evidence in experimental animals. Manifold in vitro studies clearly indicated that FA is genotoxic. FA induced various genotoxic effects in proliferating cultured mammalian cells. The cytokinesis-block micronucleus (CBMN) assay was originally developped as an ideal system form easuring micronucleus (MN), however it can also be used to measure nucleoplasmic bridges (NBP) and nuclear buds (NBUD). Over the past decade another unique mechanism of micronucleus formation, known as nuclear budding has emerged. NBUDS is considered as a marker of gene amplification and/or altered gene dosage because the nuclear budding process is the mechanism by which cells removed amplified and/excess DNA.

Formaldehyde's genotoxicity effects in pathology anatomy technologists and medical pathologists

Formaldehyde (FA) had been considered to be carcinogenic by the International Agency for Research on Cancer (group1), on the basis of sufficient evidence both in humans and in experimental animals, making it a subject of major environmental concern, especially in the occupational context. Manifold in vitro studies clearly indicated that FA is genotoxic, inducing various genotoxic effects in proliferating cultured mammalian cells. Cytokinesis-blocked micronucleus (CBMN) assay is used extensively in molecular epidemiology, and the chromosomal alterations most reported and studied by the CBMN are: micronucleus (MN), nucleoplasmic bridges (NPB) and nuclear buds (NBUDs). The pathology anatomy laboratories are work places that manipulate routinely FA and pathology anatomy technologists and pathologists contact daily with this chemical compound particularly in the macroscopic exam and grossing procedures. The aim of this study was to identify genotoxicity biomarkers in the set workers groups, such as micronucleus (MN), nucleoplasmic bridges (NPB) and nuclear buds (NBUD) in peripheral blood lymphocytes.

Micronuclei in peripheral blood lymphocytes in formaldehyde occupationally exposed workers

Formaldehyde (CH2O) the most simple and reactive of all aldehydes, is a colorless, reactive and readily polymerizing gas at normal temperature. It has a pungent, suffocating odour that is recognized by most human subjects at concentrations below 1 ppm. According to the Report on Carcinogens, formaldehyde (FA) ranks 25th in the overall U.S. chemical production with more than 11 billion pounds (5 million tons) produced each year. Is an important industrial compound that is used in the manufacture of synthetic resins and chemical compounds such as lubricants and adhesives. It has also applications as a disinfectant, preservative and is used in cosmetics. Estimates of the number of persons who are occupationally exposed to FA indicate that, at least at low levels, may occur in a wide variety of industries. The occupational settings with most extensive use of formaldehyde is in the production of resins and in anatomy and pathology laboratories. Several studies reported a carcinogenic effect in humans after inhalation of FA, in particular an increased risk for nasopharyngeal cancer. Nowadays, the International Agency for Research on Cancer (IARC) classifies FA as carcinogenic to humans (group 1), on the basis of sufficient evidence in humans and sufficient evidence in experimental animals. Manifold in vitro studies clearly indicated that FA is genotoxic. FA induced various genotoxic effects in proliferatin cultured mammalian cells. A variety of evidence suggests that the primary DNA alterations after FA exposure are DNA-protein crosslinks (DPX). Incomplete repair of DPX can lead to the formation of mutations.

Exposição ocupacional a formaldeído: avaliação da exposição e efeitos genotóxicos

O formaldeído (FA) foi classificado, em 2004, pela International Agency for Cancer Research como agente cancerígeno. Este agente químico ocupa a 25ª posição em toda a produção química dos Estados Unidos da América, com mais de 5 milhões de toneladas produzidas por ano. Devido à sua importância económica e uso diversificado, muitos indivíduos estão expostos profissionalmente a FA. Com o estudo desenvolvido pretendeu‑se avaliar a exposição a FA em dois contextos ocupacionais distintos – na produção de FA e resinas e em laboratórios de anatomia patológica (AP) e relacionar com eventuais efeitos para a saúde, comparando a frequência de micronúcleos (MN) em linfócitos do sangue periférico e em células esfoliadas da mucosa bucal dos trabalhadores expostos a FA com indivíduos não expostos (controlos). Como amostra foram estudados 80 trabalhadores ocupacionalmente expostos a FA: 30 trabalhadores da fábrica de produção de FA e resinas e 50 trabalhadores de 10 laboratórios de AP. Foi constituído um grupo controlo de 85 indivíduos com atividades profissionais que não envolviam a exposição a formaldeído ou qualquer outro agente químico com propriedades genotóxicas. Aplicaram‑se duas metodologias distintas de avaliação ambiental do FA com o objetivo de conhecer a exposição profissional. Compararam‑se os resultados obtidos com os valores limite para a exposição média ponderada (TLV‑TWA=0,75 ppm) e para a concentração máxima (VLE‑CM=0,3 ppm). A totalidade dos laboratórios apresentou resultados superiores ao valor de referência existente para a concentração máxima. Nenhum dos resultados obtidos para a exposição média ponderada foi superior ao valor de referência. O exame macroscópico obteve os valores das concentrações máximas mais elevadas em 90% dos laboratórios. Os valores de MN foram mais elevados nos indivíduos expostos a FA comparativamente com os controlos. No caso dos MN nos linfócitos, a média foi de 3,96 nos expostos e de 0,81 nos não expostos. Os MN nas células esfoliadas da boca apresentaram uma média de 0,96 nos expostos e de 0,16 nos controlos. Os resultados obtidos nesta acção de biomonitorização podem revelar‑se particularmente úteis para as organizações responsáveis em definir os níveis aceitáveis para a exposição humana a FA. ABSTRACT: Since 2004, formaldehyde (FA) has been classified by the International Agency for Cancer Research as a carcinogen. The FA ranks 25th in the overall United States chemical production, with more than 5 million tons produced each year. Due to its economic importance and varied use, many individuals are exposed to FA at their occupational settings. This study aimed to assess the exposure to FA in two occupational settings – FA production factory and pathology anatomy (PA) laboratories – and relate it to possible health effects by comparing frequency of micronuclei (MN) in peripheral blood lymphocytes and exfoliated cells from the oral mucosa of workers exposed to FA with individuals not exposed to this agent (controls). This study was performed in 80 workers occupationally exposed to FA: 30 workers of the FA factory and 50 workers in 10 PA laboratories. The control group comprised 85 subjects without exposure. We have applied two different methodologies for environmental monitoring of FA. The results were compared with the reference to the exposure weighted average (TLV‑TWA = 0.75 ppm) and ceiling concentration (VLE‑MC = 0.3 ppm). All laboratories had results higher than the reference value to CM (1.41 ppm). None of the results obtained for the TWA exposure (0.16 ppm) were higher than the reference value. Macroscopic examination obtained the highest values of CM in 90% of laboratories. MN values were higher in individuals exposed to FA as compared to controls. As for MN in lymphocytes, the average was 3.96 in exposed compared with 0.81 in the unexposed. The MN in exfoliated cells of the buccal mucosa had an average of 0.96 in exposed, compared with 0.16 in controls. The results of this biomonitoring can be particularly useful to organizations responsible for defining acceptable levels for human exposure to FA.

Supercritical carbon dioxide extraction of bioactive compounds from microalgae and volatile oils from aromatic plants

A discussion of the most interesting results obtained in our laboratories, during the supercritical CO(2) extraction of bioactive compounds from microalgae and volatile oils from aromatic plants, was carried out. Concerning the microalgae, the studies on Botryococcus braunii and Chlorella vulgaris were selected. Hydrocarbons from the first microalgae, which are mainly linear alkadienes (C(23)-C(31)) with an odd number of carbon atoms, were selectively extracted at 313 K increasing the pressure up to 30.0 MPa. These hydrocarbons are easily extracted at this pressure, since they are located outside the cellular walls. The extraction of carotenoids, mainly canthaxanthin and astaxanthin, from C. vulgaris is more difficult. The extraction yield of these components at 313 K and 35.0 MPa increased with the degree of crushing of the microalga, since they are not extracellular. On the other hand, for the extraction of volatile oils from aromatic plants, studies on Mentha pulegium and Satureja montana L were chosen. For the first aromatic plant, the composition of the volatile and essential oils was similar, the main components being the pulegone and menthone. However, this volatile oil contained small amounts of waxes, which content decreased with decreasing particle size of the plant matrix. For S. montana L it was also observed that both oils have a similar composition, the main components being carvacrol and thymol. The main difference is the relative amount of thymoquinone, which content can be 15 times higher in volatile oil. This oxygenated monoterpene has important biological activities. Moreover, experimental studies on anticholinesterase activity of supercritical extracts of S. montana were also carried out. The supercritical nonvolatile fraction, which presented the highest content of the protocatechuic, vanilic, chlorogenic and (+)-catechin acids, is the most promising inhibitor of the enzyme butyrylcholinesterase. In contrast, the Soxhlet acetone extract did not affect the activity of this enzyme at the concentrations tested. (C) 2011 Elsevier B.V. All rights reserved.

Re-evaluation of a reported increased micronucleus frequency in lymphocytes of workers occupationally exposed to formaldehyde

A replicate evaluation of increased micronucleus (MN) frequencies in peripheral lymphocytes of workers occupationally exposed to formaldehyde (FA) was undertaken to verify the observed effect and to determine scoring variability. May–Grünwald–Giemsa-stained slides were obtained from a previously performed cytokinesis-block micronucleus test (CBMNT) with 56 workers in anatomy and pathology laboratories and 85 controls. The first evaluation by one scorer (scorer 1) had led to a highly significant difference between workers and controls (3.96 vs 0.81 MN per 1000 cells). The slides were coded before re-evaluation and the code was broken after the complete re-evaluation of the study. A total of 1000 binucleated cells (BNC) were analysed per subject and the frequency of MN (in ‰) was determined. Slides were distributed equally and randomly between two scorers, so that the scorers had no knowledge of the exposure status. Scorer 2 (32 exposed, 36 controls) measured increased MN frequencies in exposed workers (9.88 vs 6.81). Statistical analysis with the two-sample Wilcoxon test indicated that this difference was not significant (p = 0.17). Scorer 3 (20 exposed, 46 controls) obtained a similar result, but slightly higher values for the comparison of exposed and controls (19.0 vs 12.89; p = 0.089). Combining the results of the two scorers (13.38 vs 10.22), a significant difference between exposed and controls (p = 0.028) was obtained when the stratified Wilcoxon test with the scorers as strata was applied. Interestingly, the re-evaluation of the slides led to clearly higher MN frequencies for exposed and controls compared with the first evaluation. Bland–Altman plots indicated that the agreement between the measurements of the different scorers was very poor, as shown by mean differences of 5.9 between scorer 1 and scorer 2 and 13.0 between scorer 1 and scorer 3. Calculation of the intra-class correlation coefficient (ICC) revealed that all scorer comparisons in this study were far from acceptable for the reliability of this assay. Possible implications for the use of the CBMNT in human biomonitoring studies are discussed.

Exposure and genotoxicity assessment methodologies: the case of formaldehyde occupational exposure

Formaldehyde (FA) ranks 25th in the overall U.S. chemical production, with more than 5 million tons produced each year. Given its economic importance and widespread use, many people are exposed to FA occupationally. Recently, based on the correlation with nasopharyngeal cancer in humans, the International Agency for Research on Cancer (IARC) confirmed the classification of FA as a Group I substance. Considering the epidemiological evidence of a potential association with leukemia, the IARC has concluded that FA can cause this lymphoproliferative disorder. Our group has developed a method to assess the exposure and genotoxicity effects of FA in two different occupational settings, namely FAbased resins production and pathology and anatomy laboratories. For exposure assessment we applied simultaneously two different techniques of air monitoring: NIOSH Method 2541 and Photo Ionization Detection Equipment with simultaneously video recording. Genotoxicity effects were measured by cytokinesis-blocked micronucleus assay in peripheral blood lymphocytes and by micronucleus test in exfoliated oral cavity epithelial cells, both considered target cells. The two exposure assessment techniques show that in the two occupational settings peak exposures are still occurring. There was a statistical significant increase in the micronucleus mean of epithelial cells and peripheral lymphocytes of exposed individuals compared with controls. In conclusion, the exposure and genotoxicity effects assessment methodologies developed by us allowed to determine that these two occupational settings promote exposure to high peak FA concentrations and an increase in the micronucleus mean of exposed workers. Moreover, the developed techniques showed promising results and could be used to confirm and extend the results obtained by the analytical techniques currently available.

Classification and staging of chronic liver disease from multimodal data

Chronic liver disease (CLD) is most of the time an asymptomatic, progressive, and ultimately potentially fatal disease. In this study, an automatic hierarchical procedure to stage CLD using ultrasound images, laboratory tests, and clinical records are described. The first stage of the proposed method, called clinical based classifier (CBC), discriminates healthy from pathologic conditions. When nonhealthy conditions are detected, the method refines the results in three exclusive pathologies in a hierarchical basis: 1) chronic hepatitis; 2) compensated cirrhosis; and 3) decompensated cirrhosis. The features used as well as the classifiers (Bayes, Parzen, support vector machine, and k-nearest neighbor) are optimally selected for each stage. A large multimodal feature database was specifically built for this study containing 30 chronic hepatitis cases, 34 compensated cirrhosis cases, and 36 decompensated cirrhosis cases, all validated after histopathologic analysis by liver biopsy. The CBC classification scheme outperformed the nonhierachical one against all scheme, achieving an overall accuracy of 98.67% for the normal detector, 87.45% for the chronic hepatitis detector, and 95.71% for the cirrhosis detector.