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The data of a patient with carbamate pesticide poisoning were analyzed. Cardiac arrest, oliguria, acute renal injury and pulmonary infection occurred during treatment. After cardiopulmonary resuscitation, tracheal intubation, CRRT, anti-infection and other symptomatic support treatment, the patient recovered and discharged. The myocardial damage caused by carbamate pesticide poisoning is easy to be ignored, and it often causes cardiac manifestations such as arrhythmia and cardiac insufficiency, and the related markers of cardiac injury, electrocardiogram and echocardiogram are also changed. Therefore, the awareness of cardiac damage caused by carbamate pesticide poisoning should be improved.
Subject(s)
Humans , Pesticides , Carbamates , Heart Arrest , Arrhythmias, Cardiac , Poisoning/therapy , Organophosphate PoisoningABSTRACT
Wax apple (Syzygium samarangense) has received growing research interest for its high nutritional and medicinal value due to its constituents such as polysaccharide, organic acids, flavonoids, minerals, and other substances. In this study, wax apple polysaccharide (WAP) was isolated from this plant and its protective effect against ethyl carbamate (EC)-induced oxidative damage was evaluated in human hepatocytes (L02 cells). Firstly, a series of analyses such as high-performance liquid chromatography (HPLC), high-performance gel permeation chromatography (HPGPC), Fourier transform infrared spectroscopy (FT-IR), gas chromatography/mass spectrometry (GC/MS), and 1H and 13C nuclear magnetic resonance (NMR) were conducted to identify the structure of WAP. Thereafter, in vitro cell experiments were performed to verify the protective effects of WAP against EC-induced cytotoxicity, genotoxicity, and oxidative damage in L02 cells. Our results revealed that WAP is composed of mannose, rhamnose, glucuronic acid, galacturonic acid, glucose, galactose, arabinose, and fucose in a molar ratio of 2.20:3.94:4.45:8.56:8.86:30.82:39.78:1.48. Using a combination of methylation and NMR spectroscopic analysis, the primary structure of WAP was identified as Araf-(1→, Glcp-(1→, →2)-Araf-(1→, →3)-Galp-(1→, →3)-Araf-(1→, and →6)-Galp-(1→. Cell experiments indicated that WAP exhibited significant protective effects on EC-treated L02 cells via suppressing cytotoxicity and genotoxicity, reducing reactive oxygen species (ROS) and O2•- formation, as well as improving mitochondrial membrane potential (MMP) and glutathione (GSH). In a nutshell, WAP has the potential as an important therapeutic agent or supplement for hepatic oxidative damage. Meanwhile, further studies are needed to prove the above effects in vivo at the biological and clinical levels.
Subject(s)
Humans , Syzygium/chemistry , Urethane/pharmacology , Spectroscopy, Fourier Transform Infrared , Oxidative Stress , Glutathione/pharmacology , Hepatocytes , Polysaccharides/pharmacologyABSTRACT
Esta pesquisa buscou analisar os casos de intoxicações por organofosforados e carbamatos, presentes em agrotóxicos e pesticidas, registrados na base de dados do Centro de Informação e Assistência Toxicológica do Paraná (CIATox/PR). Trata-se de estudo retrospectivo em que foram avaliadas variáveis sociodemográficas e clínicas relacionadas à intoxicação, à forma de exposição, à causa e ao desfecho. Para a análise estatística foram utilizados testes de Kolmogorov-Smirnov, Shapiro-Wilk, Qui-Quadrado e Teste Exato de Fisher, considerando p<0,05. A amostra foi composta por 426 casos, sendo 218 (51,2%) mulheres. O principal agente foi raticida clandestino em 278 (65,2%) pacientes. A principal circunstância das intoxicações foi tentativa de suicídio com 293 (68,8%) casos. Houve predominância de exposição via oral (89,9%) e zona urbana em 349 (81,9%) casos, a maioria considerados leves (52,6%) e assistidos em serviços médicos (66,4%). Cerca de 327 (76,8%) indivíduos apresentavam manifestações clínicas. O tempo decorrido entre a exposição e o contato com o CIATox/PR foi maior nos casos considerados graves (p= 0,041). A atropina foi utilizada em 94 (49%) pacientes intoxicados por carbamatos, em 31 (33,3%) por organofosforados e em 84 (59,6%) por outros inibidores da colinesterase não especificados. Duas mulheres e um adolescente apresentaram desfecho fatal, tendo em comum a ingesta intencional de "chumbinho". A casuística predominante nesta série histórica foi paciente do sexo feminino, faixa etária adulta, residente na região metropolitana, exposta a agrotóxico por via oral, sintomática, classificada como gravidade leve, sem necessidade de atropina, contudo ocorreram três casos com desfecho para o óbito.
This research aimed to analyze the cases of poisoning by organophosphates and carbamates, present in agrochemicals and pesticides, registered in the database of the Information and Toxicological Assistance Center of Paraná (CIATox/PR). This is a retrospective study in which sociodemographic and clinical variables related to intoxication, form of exposure, cause and outcome were evaluated. For the statistical analysis, the Kolmogorov-Smirnov, Shapiro-Wilk, Chi-Square and Fisher's Exact Test were used, considering p<0.05. The sample consisted of 426 cases, 218 (51.2%) of which were women. The main agent was clandestine rodenticide in 278 (65.2%) patients. The main circumstance of intoxication was a suicide attempt with 293 (68.8%) cases. There was a predominance of oral exposure (89.9%) and urban areas in 349 (81.9%) cases, most considered mild (52.6%) and assisted in medical services (66.4%). About 327 (76.8%) individuals had clinical manifestations. The time elapsed between exposure and contact with CIATox/PR was longer in cases considered severe (p=0.041). Atropine was used in 94 (49%) patients intoxicated by carbamates, in 31 (33.3%) by organophosphates and in 84 (59.6%) by other unspecified cholinesterase inhibitors. Two women and one teenager had a fatal outcome, having in common the intentional ingestion of "chumbinho". The predominant casuistry in this historical series was a female patient, adult age, living in the metropolitan region, exposed to pesticides orally, symptomatic, classified as mild severity, without the need for atropine, however, there were three cases with an outcome of death.
Subject(s)
Humans , Male , Female , Organophosphates , Poisoning , Carbamates , Cholinesterase Inhibitors , Cholinesterases , Agrochemicals , Casuistry , RodenticidesABSTRACT
OBJECTIVES@#To evaluate the composition of bioactive substances and the antioxidant effects of jujube fruit under gut microbiota fermentation (GMF), and the inhibitory effect on cytotoxicity caused by ethyl carbamate (EC).@*METHODS@#Changes in the contents of flavonoids, polyphenols, total sugars, and reducing sugars of jujube fruit after GMF (0, 2, 6, 12, 24, and 48 h) were determined. The oxidation resistance of fermented jujube fruits (from 0 to 48 h fermentation) was evaluated using in vitro 2,2'-azinobis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS) and ferric reducing antioxidant power (FRAP) assays. Inhibitory effects of 48 h-fermented jujube fruit at various concentrations (0.25, 0.50, 1.00, and 2.00 mg/mL) on EC-treated toxicity and DNA damage of Caco-2 cells were estimated using 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) and nuclear staining assays, respectively. Effects of different concentrations of jujube fruit on EC-treated Caco-2 cells' intracellular reactive oxygen species (ROS), glutathione (GSH) levels, and mitochondrial membrane potential (MMP) were also evaluated.@*RESULTS@#Jujube fruit has rich bioactive components after GMF and shows strong antioxidant capacity. Fermented jujube fruit can inhibit the cytotoxicity and DNA damage of Caco-2 cells caused by EC and reduce intracellular ROS generation, as well as restoring GSH and MMP.@*CONCLUSIONS@#Fermented jujube fruit extracts produced by GMF still contain biologically active substances which retain biological activity and antioxidation capabilities.
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Methocarbamol is a central nervous system depressant with skeletal muscle relaxant properties indicated to treatspasms. During the synthesis of methocarbamol from guaifenesin, both potential known and an unknown impurity(0.05%–0.1%), high performance liquid chromatography) were observed. The later was separated using preparativeliquid chromatography. Upon the findings of the 1H nuclear magnetic resonance, Mass, and IR spectral analysis, theunknown impurity was designated as a β-isomer of methocarbamol [1-hydroxy-3-(2-methoxy phenoxy) propan-2-ylcarbamate]. The impurity isolation, its structure elucidation, and the potential formation mechanism are discussed.
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Ethyl carbamate (EC) is a carcinogen detected in fermented foods and alcohol beverages. Excessive intake of EC is possibly harmful to health. Enzymatic degradation is one of the most effective approaches for reducing EC in fermented foods. Urease catalyzes the hydrolysis of both EC and urea. This confers urease a good application prospect in reducing EC and its precursor urea in fermented foods. Currently, degradation of EC in alcohol beverages by urease is inefficient due to its low urethanase activity and poor affinity to EC. Urease from Bacillus amyloliquefaciens JP-21 was successfully expressed in Escherichia coli at the level of 3 292 U/L urease and 227.3 U/L urethanase. Two key residues M326 and M374 were characterized that might block the binding of enzyme to EC, through simulating docking the structure of catalytic subunit UreC of urease with EC. Three mutants (M374A, M374T and M326V) of urease with improved urethanase activity were obtained by performing point saturated mutagenesis approach. Using EC as the substrate, Km values of M374A, M374T and M326V were detected to be 101.8 mmol/L, 129.5 mmol/L and 121.7 mmol/L, respectively, which were decreased by 37.47%-50.82% compared with that of the wild type urease. These mutants can degrade more than 97% of urea in rice wine and mutant M374T shows the highest degradation of EC in rice wine. EC content in rice wine was reduced from 525 μg/L to 393 μg/L by using M374T, and the EC degradation rate of it is 0.97 folds higher than that of the wild type urease. The results are of great significance for engineering the catalytic properties of urease and improving its industrial properties, and lays a good foundation for developing strategies to reducing microbial metabolic ammonia (amine) hazards in fermented foods.
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Abstract Background: The aquatic environment is continuously being contaminated with toxic chemical substances from industrial, agricultural and domestic activities. The identification of different biomarkers of such contamination is of great relevance to determine the potential impact of those xenobiotics on the health of organisms. Objectives: To estimate the suitability of different biochemical brain biomarkers from tench in order to evaluate the environmental effect of an exposition to different concentrations of the pesticide carbofuran. Methods: The present study focused on the effect of a long-term exposure (28 days) to different concentrations (50, 100 and 200 μg/L) of the pesticide carbofuran on certain biochemical parameters of tench (Tinca tinca L.). Selected biomarkers for stress monitoring were malondialdehyde (MDA), as index of lipid peroxidation, and reduced glutathione (GSH). Moreover, acetylcholinesterase (AChE) activity was quantified as biomarker of effect on fish nervous system. Results: AChE activity was significantly inhibited in fish exposed to all concentrations of carbofuran after 14 and 21 days of exposure, returning to basal levels after this period with 50 and 100 μg/L. Moreover, significant decreases of MDA levels could be detected after 14 and 21 days of exposure to 200 μg/L, lasting until the end of the experiment. GSH levels decreased after 14 and 21 days of exposure to 100 and 200 μg/L; nevertheless, GSH recovered basal levels at the end of the experiment. Conclusion: All the three biochemical parameters assessed -mainly inhibition of AChE activity- could be adequate biomarkers of early exposition (less than 14 days) to low doses of carbofuran in biomonitoring programs, being MDA reduction also related to longer exposure.
Resumen Antecedentes: El ambiente acuático está continuamente contaminado con sustancias químicas tóxicas provenientes de actividades industriales, agrícolas y domésticas. La identificación de diferentes biomarcadores de dicha contaminación es de gran relevancia para determinar el impacto potencial de esos xenobióticos en la salud de los organismos. Objetivo: Estimar la idoneidad de diferentes biomarcadores cerebrales bioquímicos de tenca para evaluar el efecto ambiental de una exposición a diferentes concentraciones del pesticida carbofurano. Método: El presente estudio se centró en los efectos de una exposición a largo plazo (28 días) a diferentes concentraciones (50, 100 y 200 μg/L) del pesticida carbofurano sobre determinados parámetros bioquímicos de la tenca (Tinca tinca L.). Los biomarcadores seleccionados para el control del estrés fueron el malondialdehído (MDA), como indicador de peroxidación lipídica, y el glutatión reducido (GSH). Además, se cuantificó la actividad acetilcolinesterasa (AChE), como biomarcador de efecto sobre el sistema nervioso del pez. Resultados: La actividad AChE se inhibió significativamente en los peces expuestos a todas las concentraciones de carbofurano después de 14 y 21 días de exposición, regresando a los niveles basales tras este periodo en las exposiciones con 50 y 100 μg/L. Así mismo se detectaron descensos significativos de los niveles de MDA tras 14 y 21 días de exposición con 200 μg/L, manteniéndose hasta el final del experimento. Los niveles de GSH descendieron tras 14 y 21 días de exposición con 100 y 200 µg/l; sin embargo, los niveles basales se recuperaron al final del experimento. Conclusión: Los tres parámetros bioquímicos evaluados, pero principalmente la inhibición de la actividad de AChE, podrían usarse como biomarcadores de exposición temprana (menos de 14 días) a bajas dosis de carbofurano en programas de biomonitorización, estando la reducción de MDA también relacionada con exposiciones más prolongadas.
Resumo Antecedentes: O meio aquático está sendo continuamente contaminado com substâncias químicas tóxicas de atividades industriais, agrícolas e domésticas. A identificação de diferentes biomarcadores de tal contaminação é de grande relevância para determinar o impacto potencial desses xenobióticos na saúde dos organismos. Objetivo: Estimativa da idoneidade de várias biomarcadores cerebrais bioquímicos de tenca para avaliar o efeito ambiental de uma exposição a diferentes concentrações do pesticida carbofurano. Método: O presente estudo centrou-se no efeito de uma exposição a longo prazo (28 dias) a diferentes concentrações (50, 100 e 200 μg/L) do pesticida carbofurano em certos parâmetros bioquímicos da tenca (Tinca tinca L.). Os biomarcadores selecionados para o monitoramento do estresse foram malondialdeído (MDA), como índice de peroxidação lipídica e glutationa reduzida (GSH). Além disso, a atividade da acetilcolinesterase (AChE) foi quantificada, como biomarcador de efeito sobre o sistema nervoso do peixe. Resultados: A atividade de AChE foi significativamente inibida em peixes expostos a todas as concentrações de carbofurano após 14 e 21 dias de exposição, voltando aos níveis basais após esse período com 50 e 100 μg/L. Além disso, as diminuições significativas dos níveis de MDA podem ser detectadas após 14 e 21 dias de exposição a 200 μg/L, até o final do experimento. Os níveis de GSH diminuíram após 14 e 21 dias de exposição a 100 e 200 μg/L, porém GSH recuperou os níveis basais no final do experimento. Conclusão: Todos os três parâmetros bioquímicos avaliados, mas principalmente a inibição da atividade AChE, podem ser biomarcadores adequados de exposição precoce (menos de 14 dias) a baixas doses de carbofurano em programas de biomonitorização, sendo a redução de MDA também relacionada a exposições mais longas.
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ABSTRACT: The objective of this study was to review technological and toxicological factors related to presence of carbonyl compounds found in wines, including acetaldehyde, formaldehyde, acrolein, ethyl carbamate (EC) and furfural. Acetaldehyde and formaldehyde may be formed through the ethanol and methanol oxidation, respectively. Acrolein may arise as a thermal degradation product of glycerol, amino acids, carbohydrates and triglycerides or by metabolic activity of microorganisms. In addition, acrolein and furfural are formed during wood combustion; therefore, these aldehydes may be present in raw materials due to the environmental contamination. Furfural is also a product of the Maillard reaction formed from sugars and amino acids, while ethyl carbamate occurs through the reaction between urea and ethanol. These compounds may react with SO2 and phenolic compounds to form non-volatile adducts, which positively modulates color stability, astringency and aroma in wine. However, when ingested through wine, electrophilic carbonyl compounds may form adducts with nucleophilic targets, such as DNA, resulting in genotoxicity along the gastrointestinal tract. Furthermore, carbonyl compounds induce the increase of reactive oxygen species and can trigger apoptosis, in addition to hepatocellular adenoma and carcinoma as a consequence of chronic hepatotoxicity. Neurodegenerative diseases may be related to the exposure to carbonyl compounds. Therefore, strategies to reduce the levels of these compounds should be studied in order to get the most out of the beneficial functional properties of wine consumption.
RESUMO: O objetivo deste estudo foi revisar os fatores tecnológicos e toxicológicos relacionados à presença de compostos carbonílicos encontrados em vinhos, incluindo acetaldeído, formaldeído, acroleína, carbamato de etila (CE) e furfural. O acetaldeído e o formaldeído podem ser formados através da oxidação do etanol e do metanol, respectivamente. A acroleína pode surgir como um produto de degradação térmica de glicerol, aminoácidos, carboidratos e triglicerídeos ou pela atividade metabólica de microorganismos. Além disso, a acroleína e o furfural são formados durante a combustão da madeira. Portanto, esses aldeídos podem estar presentes nas matérias-primas devido à contaminação ambiental. O furfural é também um produto da reação de Maillard formado a partir de açúcares e aminoácidos, enquanto o carbamato de etila ocorre através da reação entre uréia e etanol. Estes compostos podem reagir com SO2 e compostos fenólicos para formar adutos não voláteis, que modulam positivamente a estabilidade da cor, adstringência e aroma no vinho. No entanto, quando ingeridos através do vinho, os compostos carbonílicos que são eletrofílicos podem formar adutos com alvos nucleofílicos, como o DNA, resultando em genotoxicidade ao longo do trato gastrintestinal. Além disso, os compostos carbonílicos também induzem o aumento de espécies reativas de oxigênio e podem desencadear a apoptose, além de adenoma e carcinoma hepatocelular como consequência da hepatotoxicidade crônica. Doenças neurodegenerativas podem estar relacionadas à exposição aos compostos carbonílicos. Com isso, estratégias para reduzir os níveis desses compostos devem ser estudadas para obter o máximo das propriedades funcionais benéficas do consumo de vinho.
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OBJECTIVE： To evaluate the DNA damage response of procarbazine hydrochloride （PCZ） and ethyl carbamate （EC） to different tissues in rats by performing alkaline comet assay， to validate the feasibility of alkaline comet assay of various tissues. METHODS： Totally 30 SD rats were randomly divided into 6 groups according to body weight， with 5 rats in each group， such as negative control group （hyperpure water）， PCZ 75 mg/kg group， PCZ 150 mg/kg group， EC 400 mg/kg group， EC 800 mg/kg group， positive control group （N-ethyl-N-nitrosourea 40 mg/kg）. Those rats were given relevant medicine intragasttrically for 4 d； clinical symptoms of rats were observed and body weight was recorded during experiment. Within 3 h after last medication， the rats were sacrificed； liver， renal and lung weight were weighed； liver， kidney， lung and peripheral blood lymphocytes were collected. The single cell suspension was prepared to perform alkaline comet assay. After lysis， unwind， electrophoresis and dying， tail DNA％ and tail distance of samples were analyzed by Komet 6.0 software. RESULTS： Compared with negative control group， body weight， liver and renal weight of rats were decreased significantly in PCZ 75 mg/kg group， PCZ 150 mg/kg group and positive control group 4 d after medication （P＜0.05 or P＜0.01）. Body weight of rats were decreased significantly in EC 800 mg/kg 4 d after medication （P＜0.05 or P＜0.01）； there was no statistical significance （P＞0.05）. Compared with negative control group， tail DNA％ and tail distance in liver， kidney and peripheral blood lymphocytes were increased significantly in PCZ 75 mg/kg group， PCZ 150 mg/kg group and positive control group （P＜0.05 or P＜0.01）； PCZ showed more significant effects on liver and lung. Tail DNA％ and tail distance of liver， kidney and peripheral blood lymphocytes were increased significantly in EC 800 mg/kg group （P＜0.05 or P＜0.01）， and tail DNA％ and tail distance of renal tissue was increased significantly in EC 400 mg/kg group （P＜0.05）. CONCLUSIONS： PCZ induced stronger DNA damage； liver and lung are the major genotoxicity target of PCZ. EC-induced DNA damage is relatively weak， and kidney is the most sensitive organ for EC-induced genotoxicity.
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Ethyl carbamate (EC), a carcinogenic and teratogenic chemical that is widely distributed in various alcoholic beverages, has attracted much attention. Microbial enzymatic degradation of EC in rice wine is always efficient and attractive. In this review, we summarize the research progress and problems of microbial enzymatic elimination of EC in rice wine from three aspects: the mechanisms of EC formation in rice wine, the research progress of acid urease, and the research progress of urethanase. Then, we propose the corresponding strategies to solve the problems: screening new urethanase with satisfied enzyme properties, food-grade expression and directed evolution of the bifunctional Fe³⁺-dependent acid urease and acid urease used in combination with urethanase to eliminate both urea and EC in rice wine.
Subject(s)
Oryza , Urea , Urease , Urethane , WineABSTRACT
BACKGROUND@#Assessment of acetylcholinesterase-inhibitor insecticide (AChEII) toxicity depends on the measurement of red blood cell acetylcholinesterase (RBC-AChE) activity. Its interpretation requires baseline values which is lacking in scientific literature. We aim to find the measures of central tendency and variation for RBC-AChE activity among dwellers of Anuradhapura, where the use and abuse of AChEIIs were rampant for the last few decades.@*METHODS@#A descriptive cross-sectional study with a community-based sampling for 100 healthy non-farmers (male:female = 1:1) was done using pre-determined selection criteria. Duplicate measurements of RBC-AChE activity were performed according to the modified Ellman procedure. Pearson's correlation and regression analysis were sort for RBC-AChE activity against its possible determinants.@*RESULTS@#RBC-AChE activity had a mean of 449.8 (SD 74.2) mU/μM Hb with a statistical power of 0.847. It was similar to values of "healthy controls" from previous Sri Lankan toxicological studies but was low against international reference value [586.1 (SD 65.1) mU/μM Hb]. None of the possible determinants showed a significant strength of relationship with RBC-AChE activity.@*CONCLUSION@#The baseline RBC-AChE activity among people of Anuradhapura is low in comparison with international reference values. This arises a need to find a causative mechanism.
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Young Adult , Acetylcholinesterase , Blood , Agriculture , Cross-Sectional Studies , Erythrocytes , Sri LankaABSTRACT
BACKGROUND@#Acetylcholinesterase inhibitor insecticides (AChEIIs) were used extensively in the agrarian region of Anuradhapura for the past few decades. As a result, the region faced a heightened risk of toxicity. Carbaryl, carbofuran, chlorpyrifos, dimethoate, and fenthion were the five hazardous AChEIIs banned from Anuradhapura in 2014. Assessment of post-ban trends in acute poisoning will reveal the impact of the ban. Data on availability and sales of remaining AChEIIs will guide towards preventive measures against related toxicities.@*METHODS@#Cross-sectional surveys were conducted at Anuradhapura district of Sri Lanka. Details related to acute AChEII poisoning were sorted from the Teaching Hospital Anuradhapura. Main insecticide vendors in Anuradhapura were surveyed to find information on availability and sales of AChEIIs. Chi-square for goodness of fit was performed for trends in acute poisoning and sales.@*RESULTS@#Hospital admissions related to acute AChEII poisoning have declined from 554 in 2013 to 272 in 2017. Deaths related to acute AChEII poisoning have declined from 27 in 2013 to 13 in 2017. Sales of all five banned AChEIIs had reduced by 100%. Sales of the remaining AChEIIs were declining, except for acephate, phenthoate, and profenofos. However, one of the top selling, most frequently abused carbosulfan, had the highest risk of toxicity. Chi-square for goodness of fit showed a significance (P < 0.001) between the trends of hospital admissions for acute AChEII poisoning and the sales related to AChEIIs.@*CONCLUSIONS@#Hospital admissions related to acute poisoning was declining along with the overall sales of remaining AChEIIs, during the post-AChEII ban period. Nevertheless, future vigilance is needed on the remaining AChEIIs to predict and prevent related toxicities.
Subject(s)
Female , Humans , Male , Agriculture , Cholinesterase Inhibitors , Classification , Poisoning , Commerce , Cross-Sectional Studies , Developing Countries , Government Regulation , Health Policy , Hospitalization , Incidence , Insecticides , Classification , Poisoning , Poisoning , Epidemiology , Mortality , Risk Factors , Sri Lanka , EpidemiologyABSTRACT
Objective To develop a method for determination of 18 organophosphorous and carbamate pesticides in human plasma by UPLC-MS/MS. Methods Following deproteinization by acetonitrile, an aliquot of the biological sample was injected into a C18 column(1.7μm 2.1×50mm) using 5mmol/L Ammonium acetate-methanol as the mobile phase with the flow rate of 0.3mL/min, the injection volume was 10μL. Electro spray ionization(ESI) Indicators source was applied and operated in positive ion mode, and multiple reaction monitoring(MRM) mode was used to quantify. Results The limits of detection(LODs) in human plasma ranged from 0.1 to 40ng/mL, and the limits of quantitation(LOQs) ranged from 0.5 to 50ng/mL. An excellent linearity was observed for these LOQs up to 50ng/mL. The average extraction recoveries were with in 64.3%~111.9%, relative standard deviation(RSD) is 3.9%~10.3%. Conclusion This method is specific, sensitive and accurate, and can be used to detect pesticides in forensic.
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Ethyl carbamate as a potential carcinogen commonly exists in traditional fermented foods and beverages. Enzymatic removal of ethyl carbamate from fermented foods and beverages is an efficient and safe method. In this study, we mutated urethanase from Lysinibacillus fusiformis SC02 on the Q328 site through computer aided design approaches. The half-life of resulting mutants Q328C and Q328V was detected to be 7.46 and 1.96 folds higher than that of the original enzyme, and Q328R presented better thermal-tolerance than the original urethanase when incubated at high temperature. The tolerance of Q328C to ethanol and acid also increased when compared with that of the original enzyme. The stability and tolerance to acid and ethanol of urethanase could be improved by modification on its Q328 site.
Subject(s)
Amidohydrolases , Genetics , Bacillaceae , Genetics , Bacterial Proteins , Genetics , Computer-Aided Design , Enzyme Stability , Ethanol , Mutagenesis, Site-Directed , Protein EngineeringABSTRACT
To find novel functional beverages from folk teas, 33 species of frequently used non-Camellia tea (plants other than Camellia) were collected and compared with Camellia tea (green tea, pu-erh tea and black tea) for the first time. Data are reported here on the quantities of 20 free amino acids (FAAs) and three purine alkaloids (measured by UHPLC), total polyphenols (measured by Folin-Ciocalteu assay), and antioxidant activity (DPPH). The total amounts of FAAs in non-Camellia tea (0.62-18.99 mg/g) are generally less than that of Camellia tea (16.55-24.99 mg/g). However, for certain FAAs, the quantities were much higher in some non-Camellia teas, such as γ-aminobutyric acid in teas from Ampelopsis grossedentata, Isodon serra and Hibiscus sabdariffa. Interestingly, theanine was detected in tea from Potentilla fruticosa (1.16±0.81 mg/g). Furthermore, the content of polyphenols in teas from A. grossedentata, Acer tataricum subsp. ginnala are significantly higher than those from Camellia tea; teas from I. serra, Pistacia chinensis and A. tataricum subsp. ginnala have remarkable antioxidant activities similar to the activities from green tea (44.23 μg/mL). Purine alkaloids (caffeine, theobromine and theophylline) were not detected in non-Camellia teas. The investigation suggest some non-Camellia teas may be great functional natural products with potential for prevention of chronic diseases and aging, by providing with abundant polyphenols, antioxidants and specific FAAs.
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Objective: To study the chemical constituents of Cremastra appendiculata and their neuroprotection. Methods: The chemical constituents were separated and purified by silica gel, Sephadex LH-20 column chromatography, ODS column chromatography, and HPLC column chromatography. Their structures were determined by NMR spectrum. Results: Applying various chromatographic separation methods, 23 compounds were obtained from the aceticether extraction of 55% ethanol extract of C. appendiculata. Based on physicochemical properties and spectroscopic analysis, 20 structures were identified as miltarine (1), 1-(4-β-D-glucopyranosyloxybenzyl)-2-isobutylmalate-4-methyl (2), 1-[4-(β-D-glucopyranosyloxy)benzyl]-4-methyl-2-benzylmalate (3), batatasin III-3-O-glucoside (4), eucomic acid (5), hesperidin (6), (4-methyl-1,3-phenylene) dicarbamic acid methyl ester (obtucarbamate A) (7), (3-meth-oxycarbonylamino-2-methyl-phenyl)-cabamic acid metylester (8), 4,4'-diphenylmethanebis(methyl) carbamate (9), 3,3'-dihydroxy-2',6'-bis(p-hydroxybenzyl)-5-methoxybibenzyl (10), 3',5-dihydroxy-2-(4-hydroxybenzyl)-3-methoxybibenzyl (11), 3,3'-dihydroxy-2-(4-hydroxybenzyl)-5-methoxybibenzyl (12), corylin (13), batatasin III (14), neobavaisoflavone (15), isobavachalcone (16), 2,6,2',6'-tetramethoxy-4,4-bis(2,3-epoxy-1-hydroxypropyl)biphenyl (17), coelonin (18), gigantol (19), and β-sitosterol (20). Conclusion: Compounds 7-9, 13, and 15-17 are obtained from the plants of Cremastra Lindl. for the first time. Compounds 5 and 6 are obtained from the plant for the first time. Compounds 1 and 17 have the good performance of neuroprotection in hydrogen peroxide model. Neuroprotective mechanism is likely to be achieved by weakening the oxidative damage.
ABSTRACT
Objective To evaluate the effects of low dose Ethyl Carbamate (EC)on the immune function of ICR mice and to provide evidences for developing food safety standard.Methods The ICR mice were divided into four groups,and three groups were treated with 0.1 7,0.83,1 .67 mg/kg·bw EC respectively and the control group was treated with distilled water only.The immune function of ICR mice was determined by five aspects,including cellular immunity,humoral immunity,mononuclear macrophages's phagocytosis,natural killer cell activity and the organ coefficients of immune organs. Results Compared with the control group,the 1 .67 mg/kg·bw EC significantly inhibited the proliferation of spleen lymphocyte,natural killer cell activity and the hemolysis plaque -forming ability induced by ConA (P <0.05 ). Conclusion EC can cause the inhibition of normal mouse's immune function.
ABSTRACT
Arapidscreeningmethodforthedeterminationofcarbamatepesticides(CBPs)residuesin vegetables by measuring acetylcholinesterase ( AChE ) inhibition rate using electrospray ionization mass spectrometry ( ESI-MS ) has been established. After pretreatment by QuEChERS method, sample solution reacts with AChE using acetylthiocholine as substrate. AChE inhibition rate was calculated by determination of the conversion of substrate to product ( thiocholine) using ESI-MS. The temperature, time and concentration conditions of enzymatic reactions have been optimized. The relationship between the concentration of 10 kinds of common CBPs and AChE inhibition rate was researched. Matrix effects of real vegetables were studied. The limit of detection ( LOD) , which was measured by 3 times of enzyme inhibition rate of pesticide-free vegetable samples, was 0. 01-0. 05 mg/kg. The results showed that the method was better than the current national standard method of china for rapid screening of pesticide residues and fully meet the requirements of maximum residue limits( MRL) for pesticides in food of national food safety standard. False positive results were avoided effectively due to its good ability of resistance matrix interference. The reliability was proved by analyzing vegetables with liquid chromatography-tandem mass spectrometry. The method is simple, rapid, sensitive, reliable, and can be used for the rapid, high-throughput screening of CBPs in vegetables.
ABSTRACT
Se realizó un estudio comparativo y experimental para determinar el nivel plasmático de colinesterasa (con la técnica de Ensayo del reactivo de Ellman) en 51 mujeres embarazadas y 52 no embarazadas, expuestas en forma crónica a insecticidas organofosforados y carbamatos en la zona del páramo merideño y que acudieron a control y/o consulta en el Hospital tipo I de Mucuchíes (Mérida) con la finalidad de establecer la prevalencia de sus efectos en la zona mencionada y la ulterior consecuencia en el producto de la gestación. Los datos se analizaron mediante el software estadístico SPSS versión 15 para Windows. Los resultados mostraron que las pacientes embarazadas con o sin antecedentes patológicos, sus niveles de colinesterasa tienden a disminuir en cierta magnitud, se encontró diferencia estadísticamente significativa entre los niveles de colinesterasa promedio de mujeres embarazadas con antecedentes con respecto a las que no tenían antecedentes, en las mujeres no embarazadas y sin antecedentes que vivían cerca de zonas de cultivo con exposición a plaguicidas el nivel promedio de colinesterasa fue menor al nivel promedio de las mujeres en edad reproductiva no expuestas y una disminución de los valores de colinesterasa promedio en las gestantes menores de 39 años de edad. Se concluyo que las mujeres embarazadas con antecedentes patológicos presentan inhibición de niveles de colinesterasa plasmática con respeto a las no embarazadas y sin antecedentes.
A study was conducted to determine comparative and experimental plasma cholinesterase level (with the assay technique of Ellman's reagent) in 51 pregnant women and 52 non-pregnant, chronically exposed to organophosphate and carbamate wilderness area in Merida and who came to control and / or reference in the type I Mucuchíes Hospital Merida state in order to establish the prevalence of its effects on the area mentioned and further result in the product of gestation. The data were analyzed using SPSS statistical software version 15 for Windows. The results showed that pregnant patients with or without pathological history cholinesterase levels tend to decrease to some extent, statistically significant difference was found between the average cholinesterase levels of pregnant women with a history over which they had no background in nonpregnant women with no history of living near farming areas with pesticide exposure cholinesterase level was lower average to average level of reproductive age women unexposed and decreased average cholinesterase values in pregnant women under 39 years of age. It was concluded that pregnant women with medical history presented inhibition of plasma cholinesterase levels with respect to the non-pregnant and no history.
Subject(s)
Humans , Female , Pregnancy , Adolescent , Adult , Pesticides/toxicity , Plasma Volume , Pregnancy , Insecticides, Organophosphate/adverse effects , Fetus/abnormalities , Carbamates/adverse effects , Public Health , Cholinesterases/analysisABSTRACT
Use of pesticides in agriculture to protect crops against destructive pests often leads to residues of these chemicals in foods. The levels of pesticide residues in food are often determined as a means of assessing appropriate use as well as the potential for human exposure to these chemicals and hence their potential damage to human health. The aim of this study was to determine the concentration of organophosphate and carbamate pesticides in maize samples collected from various markets in Lagos State and compare these values with established safety values. Maize (Zea mays L.) samples purchased from different markets in LagosState were analyzed for residues of organophosphate and carbamate pesticides. Analysis was done using gas chromatograph with mass spectrometric detector (GC-MS) after careful extraction and cleanup. It was found that all the maize samples contained residues of one or more organophosphate or carbamate pesticides. Mean concentrations ranged from 12.0 to 1565.4 μg/kg. Maximum residue limits (MRL) of some of the pesticides were exceeded in up to 10% of samples.