A state-of-the-science review of analytical methods for urinary dialkylphosphate metabolites in the assessment of exposure to organophosphate pesticides: From 2000 to 2022.

The general population and workers are exposed to organophosphate insecticides, one of the leading chemical classes of pesticides used in rural and urban areas. This paper aims to conduct an integrative review of the most used analytical methods for identifying and quantifying dialkylphosphate-which are metabolites of organophosphate insecticides-in the urine of exposed workers, discussing their advantages, limitations and applicability. Searches utilized the PubMed, the Scientific Electronic Library Online and the Brazilian Digital Library of Theses and Dissertations databases between 2000 and 2021. Twenty-five studies were selected. The extraction methods most used were liquid-liquid extraction (LLE) (36%) and solid-phase extraction (SPE) (36%), with the SPE being more economical in terms of time and amount of solvents needed, and presenting the best percentage of recovery of analytes, when compared with LLE. Nineteen studies (76%) used the gas chromatography method of separation, and among these, 12 records (63%) indicated mass spectrometry used as a detection technology (analyzer). Studies demonstrate that dialkylphosphates are sensitive and representative exposure biomarkers for environmental and occupational organophosphate exposure.

Urinary dialkylphosphate metabolites in the assessment of exposure to organophosphate pesticides: from 2000 to 2022.

The general population and workers are exposed to organophosphate insecticides, one of the leading chemical classes of pesticides used in rural and urban areas, in the control of arboviruses and agriculture. These pesticides cause environmental/occupational exposure and associated risks to human and environmental health. The objective of this study was to carry out an integrative review of epidemiological studies that identified and quantified dialkylphosphate metabolites in the urine of exposed populations, focusing on the vector control workers, discussing the application and the results found. Searches utilized the Pubmed, Scielo, and the Brazilian Digital Library of Theses and Dissertations (BDTD) databases between 2000 and 2021. From the 194 selected studies, 75 (39%) were with children/adolescents, 48 (24%) with rural workers, 36 (19%) with the general population, 27 (14%) with pregnant women, and 9 (4%) with vector control workers. The total dialkylphosphate concentrations found in the occupationally exposed population were higher than in the general population. Studies demonstrate that dialkylphosphates are sensitive and representative exposure biomarkers for environmental and occupational organophosphate exposure. The work revealed a lack of studies with vector control workers and a lack of studies in developing countries.

Cross-cultural adaptation of an environmental health measurement instrument: Brazilian version of the health-care waste management • rapid assessment tool.

BACKGROUND: Periodic assessment is one of the recommendations for improving health-care waste management worldwide. This study aimed at translating and adapting the Health-Care Waste Management - Rapid Assessment Tool (HCWM-RAT), proposed by the World Health Organization, to a Brazilian Portuguese version, and resolving its cultural and legal issues. The work focused on the evaluation of the concepts, items and semantic equivalence between the original tool and the Brazilian Portuguese version. METHODS: A cross-cultural adaptation methodology was used, including: initial translation to Brazilian Portuguese; back translation to English; syntheses of these translation versions; formation of an expert committee to achieve consensus about the preliminary version; and evaluation of the target audience's comprehension. RESULTS: Both the translated and the original versions' concepts, items and semantic equivalence are presented. The constructs in the original instrument were considered relevant and applicable to the Brazilian context. The Brazilian version of the tool has the potential to generate indicators, develop official database, feedback and subsidize political decisions at many geographical and organizational levels strengthening the Monitoring and evaluation (M&E) mechanism. Moreover, the cross-cultural translation expands the usefulness of the instrument to Portuguese-speaking countries in developing regions. CONCLUSION: The translated and original versions presented concept, item and semantic equivalence and can be applied to Brazil.

Experimental design approach in recombinant protein expression: determining medium composition and induction conditions for expression of pneumolysin from Streptococcus pneumoniae in Escherichia coli and preliminary purification process.

BACKGROUND: Streptococcus pneumoniae (S. pneumoniae) causes several serious diseases including pneumonia, septicemia and meningitis. The World Health Organization estimates that streptococcal pneumonia is the cause of approximately 1.9 million deaths of children under five years of age each year. The large number of serotypes underlying the disease spectrum, which would be reflected in the high production cost of a commercial vaccine effective to protect against all of them and the higher level of amino acid sequence conservation as compared to polysaccharide structure, has prompted us to attempt to use conserved proteins for the development of a simpler vaccine. One of the most prominent proteins is pneumolysin (Ply), present in almost all the serotypes known at the moment, which shows an effective protection against S. pneumoniae infections. RESULTS: We have cloned the pneumolysin gene from S. pneumoniae serotype 14 and studied the effects of eight variables related to medium composition and induction conditions on the soluble expression of rPly in Escherichia coli (E. coli) and a 28-4 factorial design was applied. Statistical analysis was carried out to compare the conditions used to evaluate the expression of soluble pneumolysin; rPly activity was evaluated by hemolytic activity assay and served as the main response to evaluate the proper protein expression and folding. The optimized conditions, validated by the use of triplicates, include growth until an absorbance of 0.8 (measured at 600 nm) with 0.1 mM IPTG during 4 h at 25°C in a 5 g/L yeast extract, 5 g/L tryptone, 10 g/L NaCl, 1 g/L glucose medium, with addition of 30 µg/mL kanamycin. CONCLUSIONS: This experimental design methodology allowed the development of an adequate process condition to attain high levels (250 mg/L) of soluble expression of functional rPly in E. coli, which should contribute to reduce operational costs. It was possible to recover the protein in its active form with 75% homogeneity.

Optimization of medium formulation and seed conditions for expression of mature PsaA (pneumococcal surface adhesin A) in Escherichia coli using a sequential experimental design strategy and response surface methodology.

PsaA, a candidate antigen for a vaccine against pneumonia, is well-conserved in all Streptococcus pneumoniae serotypes. A sequence of two-level experimental designs was used to evaluate medium composition and seed conditions to optimize the expression of soluble mature PsaA in E. coli. A face-centered central composite design was first used to evaluate the effects of yeast extract (5 and 23.6 g/L), tryptone (0 and 10 g/L), and glucose (1 and 10 g/L), with replicate experiments at the central point (14.3 g/L yeast extract, 5 g/L tryptone, 5.5 g/L glucose). Next, a central composite design was used to analyze the influence of NaCl concentration (0, 5, and 10 g/L) compared with potassium salts (9.4 g/L K(2)HPO(4)/2.2 g/L KH(2)PO(4)), and seed growth (7 and 16 h). Tryptone had no significant effect and was removed from the medium. Yeast extract and glucose were optimized at their intermediate concentrations, resulting in an animal-derived material-free culture medium containing 15 g/L yeast extract, 8 g/L glucose, 50 µg/mL kanamycin, and 0.4% glycerol, yielding 1 g/L rPsaA after 16 h induction at 25°C in shake flasks at 200 rpm. All the seed age and salt conditions produced similar yields, indicating that no variation had a statistically significant effect on expression. Instead of growing the seed culture for 16 h (until saturation), the process can be conducted with 7 h seed growth until the exponential phase. These results enhanced the process productivity and reduced costs, with 5 g/L NaCl being used rather than potassium salts.

Cloning and optimization of induction conditions for mature PsaA (pneumococcal surface adhesin A) expression in Escherichia coli and recombinant protein stability during long-term storage.

The gene corresponding to mature PsaA from Streptococcus pneumoniae serotype 14 was cloned into a plasmid with kanamycin resistance and without a purification tag in Escherichia coli to express high levels of the recombinant protein for large-scale production as a potential vaccine candidate or as a carrier for polysaccharide conjugation at Bio-Manguinhos/Fiocruz. The evaluation of induction conditions (IPTG concentration, temperature and time) in E. coli was accomplished by experimental design techniques to enhance the expression level of mature recombinant PsaA (rPsaA). The optimization of induction process conditions led us to perform the recombinant protein induction at 25°C for 16 h, with 0.1mM IPTG in Terrific Broth medium. At these conditions, the level of mature rPsaA expression obtained in E. coli BL21 (DE3) Star by pET28a induction with IPTG was in the range of 0.8 g/L of culture medium, with a 10-fold lower concentration of inducer than usually employed, which contributes to a less expensive process. Mature rPsaA expressed in E. coli BL21 (DE3) Star accounted for approximately 30-35% of the total protein. rPsaA purification by ion exchange allowed the production of high-purity recombinant protein without fusion tags. The results presented in this work confirm that the purified recombinant protein maintains its stability and integrity for long periods of time in various storage conditions (temperatures of 4 or -70°C using different cryoprotectors) and for at least 3 years at 4 or -70°C in PBS. The conformation of the stored protein was confirmed using circular dichroism. Mature rPsaA antigenicity was proven by anti-rPsaA mouse serum recognition through western blot analysis, and no protein degradation was detected after long periods of storage.

Influence of induction conditions on the expression of carbazole dioxygenase components (CarAa, CarAc, and CarAd) from Pseudomonas stutzeri in recombinant Escherichia coli using experimental design.

Carbazole 1,9a-dioxygenase (CarA), the first enzyme in the carbazole degradation pathway used by Pseudomonas sp., was expressed in E. coli under different conditions defined by experimental design. This enzyme depends on the coexistence of three components containing [2Fe-2S] clusters: CarAa, CarAc, and CarAd. The catalytic site is present in CarAa. The genes corresponding to components of carbazole 1,9a-dioxygenase from P. stutzeri were cloned and expressed by salt induction in E. coli BL21-SI (a host that allows the enhancement of overexpressed proteins in the soluble fraction), using the vector pDEST™14. The expression of these proteins was performed under different induction conditions (cell concentration, temperature, and time), with the help of two-level factorial design. Cell concentration at induction (measured by absorbance at 600 nm) was tested at 0.5 and 0.8. After salt induction, expression was performed at 30 and 37°C, for 4 h and 24 h. Protein expression was evaluated by densitometry analysis. Expression of CarAa was enhanced by induction at a lower cell concentration and temperature and over a longer time, according to the analysis of the experimental design results. The results were validated at Abs (ind) = 0.3, 25°C, and 24 h, at which CarAa expression was three times higher than under the standard condition. The behavior of CarAc and CarAd was the inverse, with the best co-expression condition tested being the standard one (Abs (ind) = 0.5, T = 37°C, and t = 4 h). The functionality of the proteins expressed in E. coli was confirmed by the degradation of 20 ppm carbazole.

[Assessment of exposure to BTEX in vehicle filling stations in Rio de Janeiro, Brazil, and risks to workers' health]. / Avaliação da exposição a BTEX em postos de revenda de combustíveis no Rio de Janeiro, Brasil, e os riscos à saúde do trabalhador.

Gasoline is a complex mixture of substances, including aromatic hydrocarbons such as benzene, toluene, ethylbenzene, and xylenes (BTEX). These compounds are emitted into the air, with the special relevance of benzene since it is provenly carcinogenic. The study aimed to assess BTEX concentrations in filling stations in the city of Rio de Janeiro, Brazil, and to calculate the cancer risk associated with such exposures. Two types of sampling were performed (stationary and mobile), adapted from methodology n. 1,501 (U.S. National Institute for Occupational Safety and Health) for aromatic hydrocarbons, in six filling stations in the West Zone of Rio de Janeiro. Stationary sampling was done near the fuel pumps, while mobile sampling was done in the breathing zone of the workers (station attendants) as they moved around the station. The samples were analyzed with gas chromatography flame ionization detector. The sampling results were used to calculate the health risk, using the indicators Hazard quotient (HQ) and Cancer risk (CR) to assess the possible non-carcinogenic and carcinogenic effects, respectively, in filling station workers. Environmental concentrations for the most of the BTEX compounds were below the recommended limits, except for benzene, a carcinogenic compound, which displayed concentrations far above the limits, leading to high cancer risk values. The results showed that there are health risks for filling station attendants, especially the risk of developing cancer from excessive exposure to benzene.

A gasolina é uma mistura complexa de substâncias, dentre elas estão hidrocarbonetos como o benzeno, tolueno, etilbenzeno e xilenos (BTEX), sendo esses compostos emitidos para a atmosfera, com destaque ao benzeno, por ser comprovadamente carcinogênico. Objetivamos avaliar as concentrações de BTEX no ar de postos de revenda de combustíveis no Município do Rio de Janeiro, Brasil, e calcular o risco de câncer associado a estas exposições. Foram realizados dois tipos de amostragem (fixa e móvel) adaptadas da metodologia nº 1.501 do Instituto Nacional de Segurança e Saúde Ocupacional (Estados Unidos), em seis postos de revenda de combustíveis na Zona Oeste da cidade. A amostragem fixa foi feita próxima às bombas de combustível, enquanto a móvel foi realizada através da coleta do ar na zona respiratória dos trabalhadores (frentistas), conforme estes se deslocavam pelo posto. As amostras foram analisadas por cromatografia gasosa com detecção por ionização em chama. Com os resultados das amostragens foram feitos os cálculos de risco à saúde, usando os indicadores Quociente de perigo (HQ) e Risco de câncer (CR), para avaliar os possíveis efeitos não-carcinogênicos e carcinogênicos, respectivamente, nos trabalhadores dos postos de revenda de combustíveis. As concentrações ambientais para a maioria dos compostos BTEX foram abaixo dos limites preconizados, menos para o benzeno, um composto carcinogênico, que apresentou concentrações muito acima dos limites, levando a altos valores de risco de câncer. Os resultados demonstraram que há riscos à saúde dos trabalhadores de postos de revenda de combustíveis, principalmente o risco de desenvolver câncer, devido à exposição excessiva ao benzeno.

La gasolina es una mezcla compleja de sustancias, entre ellas existen hidrocarburos como el benceno, tolueno, etilbenceno y xilenos (BTEX), emitiéndose estos compuestos a la atmósfera, donde se destaca el benceno, al tratarse de un carcinogénico comprobado. Los objetivos fueron evaluar las concentraciones de BTEX en el aire de puestos de reventa de combustibles, en el municipio de Río de Janeiro, Brasil, así como calcular el riesgo de cáncer asociado a estas exposiciones. Se realizaron dos tipos de muestra (fija y movible), adaptadas de la metodología nº 1.501 del Instituto Nacional de Seguridad y Salud Laboral (EE.UU.), en seis puestos de reventa de combustibles de la zona oeste de la ciudad. La muestra fija se realizó cerca de las bombas de combustible, mientras que la móvil se hizo mediante tomas de aire en la zona respiratoria de los trabajadores (de la gasolinera), a medida que estos se desplazaban por el lugar de trabajo. Las muestras se analizaron por cromatografía gaseosa con detección por ionización en llama. Junto a los resultados de las muestras, se realizaron cálculos de riesgo para la salud, usando los indicadores Cociente de peligro (HQ) y Riesgo de cáncer (CR), con el fin de evaluar los posibles efectos no-carcinogénicos y carcinogénicos, respectivamente, en los trabajadores de los puestos de reventa de combustibles. Las concentraciones ambientales para la mayoría de los compuestos BTEX estuvieron por debajo de los limites preconizados, menos en el caso del benceno, un compuesto carcinogénico, que presentó concentraciones muy por encima de los límites, llevando a altos valores de riesgo de cáncer. Los resultados demostraron que existen riesgos para la salud de los trabajadores de puestos de reventa de combustibles, principalmente, riesgo de desarrollar cáncer, debido a la exposición excesiva al benceno.

Teaching work social temporality in a public university and health. / A temporalidade social do trabalho docente em universidade pública e a saúde.

This study aimed to analyze teaching work's temporality aspects in their relationship with health. Qualitative research was conducted with the participation of ten professors from a public university. Six meetings called "Worker's Health workshops" were held, a research methodology that gathers workers and researchers to discuss topics related to work and health. The material was analyzed through the content analysis technique, in the thematic modality. The main themes were identified from the transcribed materials of the workshops' dialogues, namely: working time intensification and extension, teacher's health, working time and sleep deprivation, and health advocacy strategies and solutions. We noted that health advocacy strategies are located in individuals and the workgroup, through cooperation and social time. The theme of an intensive and extensive combination of working time associated with insecure work conditions was identified. We concluded that teaching work time is underpinned by rules and social values under the historical determination of new managerial standards within the public university.

[Notebooks on health and work: diaries of public university professors]. / Cadernetas de saúde e trabalho: diários de professores de universidade pública.

The main objective of this article is to present and analyze the research instrument called "health and work notebooks", focusing on the production of knowledge concerning professors' work at a public university. The notebooks serve as a qualitative and participant research technique that is appropriate for the in-depth study of relations between health and work, viewing workplaces as privileged spaces for exercising effective action in the defense of health and the work experience as the principal material for analysis. The notebooks' special quality as a research technique lies in worker's role as protagonist in the research, as the diary's author and co-participant in the study. Eight professors participated, all from the same institute in a federal university (IFES) in Rio de Janeiro, Brazil. As for analysis of the empirical materials from the notebooks and consistent with the workers, the thematic analysis technique was adopted, producing four main discussion categories: time on the job and professors' multiple work activities; precarization of working conditions at universities; faculty health at limits; and the notebooks viewed from the authors' perspective. As for the results, the theme that stood out was work overload and time pressure to meet targets. Finally, the health and work notebooks proved to be a potential research tool for generating knowledge from a collective perspective.

Este artigo tem como objetivo principal apresentar e analisar o instrumento de pesquisa designado como "cadernetas de saúde e trabalho", com foco na produção de conhecimento sobre o trabalho de docentes de universidade pública. Trata-se de uma técnica de investigação de caráter qualitativo e participativo, adequada ao aprofundamento do estudo das relações entre a saúde e o trabalho, reavendo os locais de trabalho como lugares privilegiados de observações para o exercício de uma efetiva ação de defesa da saúde e a experiência do trabalho como matéria principal de análise. A qualidade especial das cadernetas como técnica de investigação refere-se ao papel protagonista atribuído ao trabalhador no processo de pesquisa, autor do diário e coparticipante do estudo. Participaram oito docentes pertencentes ao mesmo instituto de uma universidade federal de ensino superior (IFES), localizada no Rio de Janeiro, Brasil. Quanto à análise dos materiais empíricos, procedentes das anotações das cadernetas e de encontro com os trabalhadores, adotou-se a técnica de análise temática, chegando-se a quatro categorias principais de discussão, a saber: o tempo de trabalho e as múltiplas atividades de trabalho do professor; precarização das condições de trabalho em universidades; saúde docente entre limites e cadernetas sob o olhar dos seus autores. No que concerne aos resultados, sobressaiu o tema alusivo à sobrecarga de trabalho e pressão do tempo para cumprimento de metas. Ao fim, considerou-se que as cadernetas de saúde e trabalho mostraram-se como ferramenta de pesquisa com potencial para se gerar conhecimento em perspectiva coletiva.

El objetivo principal de este artículo es presentar y analizar el instrumento de investigación denominado "cartillas de salud y trabajo", centrándose en la producción de conocimiento sobre el trabajo de docentes de universidad pública. Se trata de una técnica de investigación de carácter cualitativo y participativo, adecuada a la profundización del estudio de las relaciones entre la salud y el trabajo, reincorporando los lugares de trabajo como lugares privilegiados de observación para el ejercicio de una efectiva acción de defensa de la salud, y la experiencia laboral como materia principal de análisis. La cualidad especial de las cartillas como técnica de investigación se refiere al papel protagonista, atribuido al trabajador en el proceso de investigación, autor del diario y coparticipante del estudio. Participaron ocho docentes pertenecientes al mismo instituto de una universidad federal de enseñanza superior (IFES), localizada en Río de Janeiro, Brasil. En cuanto al análisis de los materiales empíricos, procedentes de las anotaciones de las cartillas y de encuentros con los trabajadores, se adoptó la técnica de análisis temático, llegándose a cuatro categorías principales de discusión, a saber: el tiempo de trabajo y las múltiples actividades de trabajo del profesor; precarización de las condiciones de trabajo en universidades; salud docente entre límites y cartillas bajo la atenta mirada de sus autores. En lo que concierne a los resultados, se resaltó el tema alusivo a la sobrecarga de trabajo y presión de tiempo para el cumplimiento de metas. Al final, se consideró que las cartillas de salud y trabajo se mostraron como herramientas de investigación con potencial para generar conocimiento desde una perspectiva colectiva.

Expression and homology modeling of 2-aminobiphenyl-2,3-diol-1,2-dioxygenase from Pseudomonas stutzeri carbazole degradation pathway.

The enzyme 2'-aminobiphenyl-2,3-diol-1,2-dioxygenase (CarB), encoded by two genes (carBa and carBb), is an alpha(2)beta(2) heterotetramer that presents meta-cleavage activity toward the hydroxylated aromatic ring in the carbazole degradation pathway from petroleum-degrader bacteria Pseudomonas spp. The 1,082-base pair polymerase chain reaction product corresponding to carBaBb genes from Pseudomonas stutzeri ATCC 31258 was cloned by site-specific recombination and expressed in high levels in Escherichia coli BL21-SI with a histidine-tag and in native form. The CarB activity toward 2,3-dihydroxybiphenyl was similar for these two constructions. The alpha(2)beta(2)-heterotetrameric 3D model of CarB dioxygenase was proposed by homology modeling using the protocatechuate 4,5-dioxygenase (LigAB) structure as template. Accordingly, His12, His53, and Glu230 coordinate the Fe(II) in the catalytic site at the subunit CarBb. The model also indicates that His182 is the catalytic base responsible for deprotonating one of the hydroxyl group of the substrate by a hydrogen bond. The hydrophobic residues Trp257 and Phe258 in the CarB structure substituted the LigAB amino acid residues Ser269 and Asn270. These data could explain why the CarB was active for 2,3-dihydroxybiphenyl and not for protocatechuate.

Regulation of the synthetic estrogen 17α-ethinylestradiol in water bodies in Europe, the United States, and Brazil.

The synthetic estrogen 17α-ethinylestradiol, the principal component of oral contraceptives, has been identified as one of the main compounds accounting for adverse effects on the endocrine system in various species. This study aimed to analyze the state-of-the-art in legislation and guidelines for the control of this synthetic estrogen in water bodies in Europe and the United States and to draw a parallel with the Brazilian reality. Countries have generally attempted to expand the regulation and monitoring of certain emerging micropollutants not previously covered by legislation. Europe is more advanced in terms of water quality, while in the United States this estrogen is only regulated in water for human consumption. Brazil still lacks legal provisions or standards for this estrogen, which can be explained by the relatively limited maturity of the country's system for controlling water pollutants.

Recombinant L-Asparaginase from Zymomonas mobilis: A Potential New Antileukemic Agent Produced in Escherichia coli.

L-asparaginase is an enzyme used as a chemotherapeutic agent, mainly for treating acute lymphoblastic leukemia. In this study, the gene of L-asparaginase from Zymomonas mobilis was cloned in pET vectors, fused to a histidine tag, and had its codons optimized. The L-asparaginase was expressed extracellularly and intracellularly (cytoplasmically) in Escherichia coli in far larger quantities than obtained from the microorganism of origin, and sufficient for initial cytotoxicity tests on leukemic cells. The in silico analysis of the protein from Z. mobilis indicated the presence of a signal peptide in the sequence, as well as high identity to other sequences of L-asparaginases with antileukemic activity. The protein was expressed in a bioreactor with a complex culture medium, yielding 0.13 IU/mL extracellular L-asparaginase and 3.6 IU/mL intracellular L-asparaginase after 4 h of induction with IPTG. The cytotoxicity results suggest that recombinant L-asparaginase from Z. mobilis expressed extracellularly in E.coli has a cytotoxic and cytostatic effect on leukemic cells.

Atividade estrogênica de desreguladores endócrinos em águas superficiais do município de Santa Maria Madalena, Sudeste do Brasil / Estrogenic activity of endocrine disruptors in the surface water of Santa Maria Madalena, Southeastern Brazil

RESUMO Nas últimas décadas se intensificou o lançamento, no meio hídrico, de contaminantes com capacidade de desregulação endócrina. Nesse contexto, a aplicação e o aprimoramento de métodos analíticos eficientes passam a ser cada vez mais requeridos para verificar os potenciais impactos dessas substâncias sobre os organismos a elas expostos. Bioensaios podem ser conduzidos para esse fim, como, por exemplo, o ensaio in vitro yeast estrogen screen (YES), que permite a detecção de compostos estrogênicos e citotóxicos. Este estudo, aplicando tal ensaio, investigou a atividade estrogênica das águas e seus potenciais riscos à biota aquática de dois corpos hídricos do município de Santa Maria Madalena, no estado do Rio de Janeiro, Sudeste do Brasil. Observou-se atividade estrogênica apenas no córrego São Domingos, com concentrações de equivalente estradiol (EQ-E2) de 23 e 10.4 ng.L−1 no período de estiagem e chuvoso, respectivamente. Os potenciais riscos desse nível de atividade estrogênica foram mensurados por meio de uma avaliação de risco, que revelou alto e médio risco nas estações seca e chuvosa, respectivamente. Cabe mencionar que, entre os corpos hídricos estudados, a área do córrego São Domingos é aquela com maior adensamento populacional e menor cobertura de coleta e tratamento de esgotos. O ensaio YES mostrou ser uma boa ferramenta para a detecção da presença de desreguladores endócrinos estrogênicos, que pode subsidiar o avanço do conhecimento de corpos hídricos e a formulação de legislação e regulamentações da área ambiental.

ABSTRACT In recent decades, the release of contaminants with endocrine disrupting capacity into the water environment has intensified. In this context, the application and improvement of efficient analytical methods are increasingly required to verify the potential impacts of these substances on the organisms exposed to them. Bioassays can be conducted with this purpose, such as the yeast estrogen screen (YES) in vitro assay, which allows the detection of estrogenic and cytotoxic compounds. This study, applying the YES assay, investigated the estrogenic activity and its potential risks to the aquatic biota of two streams in Santa Maria Madalena, Rio de Janeiro State, Southeastern Brazil. Estrogenic activity was observed only in São Domingos stream, with estradiol equivalent (EEQ) concentrations of 23 and 10.4 ng.L−1 during the dry and rainy season, respectively. The potential risks of this level of estrogenic activity were evaluated by a risk assessment, which revealed a high and medium risk to the local biota in the dry and rainy seasons, respectively. Among the studied water bodies, São Domingos watershed has a bigger population density and less sewage collection and treatment comparatively to Ribeirão Santíssimo stream. The YES assay presented itself as a useful tool for spotting estrogenic endocrine disruptors, with the potential to help in expanding knowledge about water bodies and to create and enforce environmental legislation and regulations.

Correction: Recombinant L-Asparaginase from Zymomonas mobilis: A Potential New Antileukemic Agent Produced in Escherichia coli.

[This corrects the article DOI: 10.1371/journal.pone.0156692.].

Avaliação da exposição a BTEX em postos de revenda de combustíveis no Rio de Janeiro, Brasil, e os riscos à saúde do trabalhador / Assessment of exposure to BTEX in vehicle filling stations in Rio de Janeiro, Brazil, and risks to workers' health / Evaluación de la exposición a BTEX en puestos de reventa de combustibles, en Río de Janeiro, Brasil, y riesgos para la salud del trabajador

A gasolina é uma mistura complexa de substâncias, dentre elas estão hidrocarbonetos como o benzeno, tolueno, etilbenzeno e xilenos (BTEX), sendo esses compostos emitidos para a atmosfera, com destaque ao benzeno, por ser comprovadamente carcinogênico. Objetivamos avaliar as concentrações de BTEX no ar de postos de revenda de combustíveis no Município do Rio de Janeiro, Brasil, e calcular o risco de câncer associado a estas exposições. Foram realizados dois tipos de amostragem (fixa e móvel) adaptadas da metodologia nº 1.501 do Instituto Nacional de Segurança e Saúde Ocupacional (Estados Unidos), em seis postos de revenda de combustíveis na Zona Oeste da cidade. A amostragem fixa foi feita próxima às bombas de combustível, enquanto a móvel foi realizada através da coleta do ar na zona respiratória dos trabalhadores (frentistas), conforme estes se deslocavam pelo posto. As amostras foram analisadas por cromatografia gasosa com detecção por ionização em chama. Com os resultados das amostragens foram feitos os cálculos de risco à saúde, usando os indicadores Quociente de perigo (HQ) e Risco de câncer (CR), para avaliar os possíveis efeitos não-carcinogênicos e carcinogênicos, respectivamente, nos trabalhadores dos postos de revenda de combustíveis. As concentrações ambientais para a maioria dos compostos BTEX foram abaixo dos limites preconizados, menos para o benzeno, um composto carcinogênico, que apresentou concentrações muito acima dos limites, levando a altos valores de risco de câncer. Os resultados demonstraram que há riscos à saúde dos trabalhadores de postos de revenda de combustíveis, principalmente o risco de desenvolver câncer, devido à exposição excessiva ao benzeno.

Gasoline is a complex mixture of substances, including aromatic hydrocarbons such as benzene, toluene, ethylbenzene, and xylenes (BTEX). These compounds are emitted into the air, with the special relevance of benzene since it is provenly carcinogenic. The study aimed to assess BTEX concentrations in filling stations in the city of Rio de Janeiro, Brazil, and to calculate the cancer risk associated with such exposures. Two types of sampling were performed (stationary and mobile), adapted from methodology n. 1,501 (U.S. National Institute for Occupational Safety and Health) for aromatic hydrocarbons, in six filling stations in the West Zone of Rio de Janeiro. Stationary sampling was done near the fuel pumps, while mobile sampling was done in the breathing zone of the workers (station attendants) as they moved around the station. The samples were analyzed with gas chromatography flame ionization detector. The sampling results were used to calculate the health risk, using the indicators Hazard quotient (HQ) and Cancer risk (CR) to assess the possible non-carcinogenic and carcinogenic effects, respectively, in filling station workers. Environmental concentrations for the most of the BTEX compounds were below the recommended limits, except for benzene, a carcinogenic compound, which displayed concentrations far above the limits, leading to high cancer risk values. The results showed that there are health risks for filling station attendants, especially the risk of developing cancer from excessive exposure to benzene.

La gasolina es una mezcla compleja de sustancias, entre ellas existen hidrocarburos como el benceno, tolueno, etilbenceno y xilenos (BTEX), emitiéndose estos compuestos a la atmósfera, donde se destaca el benceno, al tratarse de un carcinogénico comprobado. Los objetivos fueron evaluar las concentraciones de BTEX en el aire de puestos de reventa de combustibles, en el municipio de Río de Janeiro, Brasil, así como calcular el riesgo de cáncer asociado a estas exposiciones. Se realizaron dos tipos de muestra (fija y movible), adaptadas de la metodología nº 1.501 del Instituto Nacional de Seguridad y Salud Laboral (EE.UU.), en seis puestos de reventa de combustibles de la zona oeste de la ciudad. La muestra fija se realizó cerca de las bombas de combustible, mientras que la móvil se hizo mediante tomas de aire en la zona respiratoria de los trabajadores (de la gasolinera), a medida que estos se desplazaban por el lugar de trabajo. Las muestras se analizaron por cromatografía gaseosa con detección por ionización en llama. Junto a los resultados de las muestras, se realizaron cálculos de riesgo para la salud, usando los indicadores Cociente de peligro (HQ) y Riesgo de cáncer (CR), con el fin de evaluar los posibles efectos no-carcinogénicos y carcinogénicos, respectivamente, en los trabajadores de los puestos de reventa de combustibles. Las concentraciones ambientales para la mayoría de los compuestos BTEX estuvieron por debajo de los limites preconizados, menos en el caso del benceno, un compuesto carcinogénico, que presentó concentraciones muy por encima de los límites, llevando a altos valores de riesgo de cáncer. Los resultados demostraron que existen riesgos para la salud de los trabajadores de puestos de reventa de combustibles, principalmente, riesgo de desarrollar cáncer, debido a la exposición excesiva al benceno.

Evaluation of the stability of concentrated emulsions for lemon beverages using sequential experimental designs.

The study of the stability of concentrated oil-in-water emulsions is imperative to provide a scientific approach for an important problem in the beverage industry, contributing to abolish the empiricism still present nowadays. The use of these emulsions would directly imply a reduction of transportation costs between production and the sales points, where dilution takes place. The goal of this research was to evaluate the influence of the main components of a lemon emulsion on its stability, aiming to maximize the concentration of oil in the beverage and to correlate its physicochemical characteristics to product stability, allowing an increase of shelf life of the final product. For this purpose, analyses of surface and interface tension, electrokinetic potential, particle size and rheological properties of the emulsions were conducted. A 2(4-1) fractional factorial design was performed with the following variables: lemon oil/water ratio (30% to 50%), starch and Arabic gum concentrations (0% to 30%) and dioctyl sodium sulfosuccinate (0 mg/L to 100 mg/L), including an evaluation of the responses at the central conditions of each variable. Sequentially, a full design was prepared to evaluate the two most influential variables obtained in the first plan, in which concentration of starch and gum ranged from 0% to 20%, while concentration of lemon oil/water ratio was fixed at 50%, without dioctyl sodium sulfosuccinate. Concentrated emulsions with stability superior to 15 days were obtained with either starch or Arabic gum and 50% lemon oil. The most stable formulations presented viscosity over 100 cP and ratio between the surface tension of the emulsion and the mucilage of over 1. These two answers were selected, since they better represent the behavior of emulsions in terms of stability and could be used as tools for an initial selection of the most promising formulations.

On the debate about teleology in biology: the notion of "teleological obstacle".

Among the epistemological obstacles described by Gaston Bachelard, we contend that unitary and pragmatic knowledge is correlated to the teleological categories of Ernst Mayr and is the basis for prevailing debate on the notion of "function" in biology. Given the proximity of the aspects highlighted by these authors, we propose to associate the role of teleological thinking in biology and the notion of unitary and pragmatic knowledge as an obstacle to scientific knowledge. Thus, teleological thinking persists acting as an epistemological obstacle in biology, according to Bachelardian terminology. Our investigation led us to formulate the "teleological obstacle," which we consider important for the future of biology and possibly other sciences.

Evaluation of pre-induction temperature, cell growth at induction and IPTG concentration on the expression of a leptospiral protein in E. coli using shaking flasks and microbioreactor.

BACKGROUND: Leptospirosis is a zoonose that is increasingly endemic in built-up areas, especially where there are communities living in precarious housing with poor or non-existent sanitation infrastructure. Leptospirosis can kill, for its symptoms are easily confused with those of other diseases. As such, a rapid diagnosis is required so it can be treated effectively. A test for leptospirosis diagnosis using Leptospira Immunoglobulin-like (Lig) proteins is currently at final validation at Fiocruz. RESULTS: In this work, the process for expression of LigB (131-645aa) in E. coli BL21 (DE3)Star™/pAE was evaluated. No significant difference was found for the experiments at two different pre-induction temperatures (28 °C and 37 °C). Then, the strain was cultivated at 37 °C until IPTG addition, followed by induction at 28°C, thereby reducing the overall process time. Under this condition, expression was assessed using central composite design for two variables: cell growth at which LigB (131-645aa) was induced (absorbance at 600 nm between 0.75 and 2.0) and inducer concentration (0.1 mM to 1 mM IPTG). Both variables influenced cell growth and protein expression. Induction at the final exponential growth phase in shaking flasks with Abs(ind) = 2.0 yielded higher cell concentrations and LigB (131-645aa) productivities. IPTG concentration had a negative effect and could be ten-fold lower than the concentration commonly used in molecular biology (1 mM), while keeping expression at similar levels and inducing less damage to cell growth. The expression of LigB (131-645aa) was associated with cell growth. The induction at the end of the exponential phase using 0.1 mM IPTG at 28 °C for 4 h was also performed in microbioreactors, reaching higher cell densities and 970 mg/L protein. LigB (131-645aa) was purified by nickel affinity chromatography with 91% homogeneity. CONCLUSIONS: It was possible to assess the effects and interactions of the induction variables on the expression of soluble LigB (131-645aa) using experimental design, with a view to improving process productivity and reducing the production costs of a rapid test for leptospirosis diagnosis.

Prevalência do tremor essencial em população exposta ocupacionalmente a agrotóxicos no Estado do Rio de Janeiro / Prevalence of essential tremor in occupationally exposed population to pesticides in the State of Rio de Janeiro

Introdução: O tremor essencial é um dos distúrbios de movimento mais frequentes em adultos. Objetivo: Estimar a prevalência de tremor essencial em um grupo de guardas de endemia expostos cronicamente a agrotóxicos, identificar seus municípios de lotação e os principais inseticidas e larvicidas aplicados. Método: Estudo transversal descritivo e inferencial com dados obtidos de prontuários médicos dos guardas de endemias oriundos da Fundação Nacional de Saúde (Funasa), que buscaram atendimento no Ambulatório de Neurotoxicologia Ocupacional e Ambiental do Cesteh/Ensp/Fiocruz no período entre agosto/2010 a setembro/2012. Resultados: Foram analisados 383 registros. A estimativa da prevalência de tremor essencial foi de 14,4% (IC95%: 10,9 - 17,9). Foram identificados 19 municípios de lotação e 12 princípios ativos aplicados. Conclusão: Observou-se elevada prevalência de tremor essencial nesta amostra. Limitações no desenho devem ser consideradas, sendo necessário avaliar seus resultados com cautela. (AU)

Introdution: Essential tremor is one of the most frequent movement disorders in adults. Objective: To estimate the prevalence of essential tremor in a group of public pesticide applicators, identify their counties and the main insecticides and larvicides applied. Method: This is a descriptive and inferential, transversal analysis performed from the secondary data obtained from medical records of public pesticide applicators originated from Fundação Nacional de Saúde (Funasa), seeking care at Outpatient Clinic of Occupational and Environmental Neurotoxicology from Cesteh/Ensp/Fiocruz between august/ 2010 and september/2012. Results: In our sample, the estimate of the prevalence of essential tremor was 14.4 % (95%CI: 10,9 to 17,9), 19 counties and 12 pesticides applied were identified. Conclusion: It was observed a high prevalence of essential tremor, but this result must be treated with caution. (AU)