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1.
J Environ Sci Health B ; 55(4): 355-360, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31868080

RESUMO

The aim of this study was to evaluate the DNA damage in soybean growers during two agricultural periods of a crop season (high and low exposure) and a control group, as well as butyrylcholinesterase (BChE) activity during these exposure periods in order to estimate the degree of BChE inhibition for the exposed group. DNA damage in peripheral whole blood was evaluated by the comet assay and plasma BChE activity was accessed as a measure of exposure to cholinesterase inhibitors. None of the soybean growers reported using full Personal Protective Equipment (PPE). BChE was lower in high exposure period than in low exposure period and DNA damage index was significantly increased in the high exposure period than in the low exposure period. In addition, DNA damage in both exposure periods was higher than control group. No correlation was found between exposure time and DNA damage and BChE activity. However, negative correlation was observed between DNA damage in high and low exposure periods. The results indicate that soybean growers are exposed to cholinesterase inhibitors and to pesticides mixtures with genotoxic potential.


Assuntos
Butirilcolinesterase/sangue , Dano ao DNA , Fazendeiros , Exposição Ocupacional/análise , Praguicidas/toxicidade , Soja , Adulto , Agricultura , Brasil , Estudos de Casos e Controles , Inibidores da Colinesterase/toxicidade , Ensaio Cometa , Humanos , Masculino , Pessoa de Meia-Idade , Fatores de Tempo
2.
Chem Commun (Camb) ; 55(97): 14574-14577, 2019 Dec 18.
Artigo em Inglês | MEDLINE | ID: mdl-31663530

RESUMO

Non-specific binding of a fluorescent probe to human serum albumin is problematic because it induces signal interference when the probe detects the target biomarker in human serum. To eliminate this problem, we used intrinsically problematic non-specific fluorescence in designing a fluorescent probe for butyrylcholinesterase activity in serum. The probe containing a fluorophore with specific binding affinity for albumin could sensitively detect butyrylcholinesterase activity in serum with high selectivity to acetylcholinesterase and screen the efficiency of butyrylcholinesterase inhibitors.


Assuntos
Butirilcolinesterase/sangue , Fluorescência , Corantes Fluorescentes/análise , Corantes Fluorescentes/química , Albumina Sérica Humana/química , Sítios de Ligação , Butirilcolinesterase/metabolismo , Inibidores da Colinesterase/química , Inibidores da Colinesterase/farmacologia , Corantes Fluorescentes/metabolismo , Galantamina/química , Galantamina/farmacologia , Humanos , Estrutura Molecular , Albumina Sérica Humana/metabolismo , Espectrometria de Fluorescência , Tacrina/química , Tacrina/farmacologia
3.
Biomarkers ; 24(8): 771-775, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31642715

RESUMO

Background: Farmers and their workers are exposed to a wide variety of pesticides. The use of pesticides has been documented to lead to several adverse health effects. Inhibition of cholinesterase, primarily butyrylcholinesterase is a good indicator of occupational exposure to organophosphates and carbamates.Objective: This case-control study aims to study the risks associated with pesticide exposure among farmers and agricultural workers in the Souss Massa region by analyzing variations in the response of a pesticides exposure biomarker: Serum Cholinesterase Activity (butyrylcholinesterase (BChE)).Materials and methods: This was a prospective study conducted on 133 participants (71 farmers and 62 non-farmers). A structured questionnaire was applied collecting socio-demographic information and determining knowledge and work practices in relation to pesticide use. The activity of Serum cholinesterase was measured by the butyrulthiocholine method a spectrophotometric assay.Results: The mean age of the participants was 42.5 ± 10.66 years. The study demonstrated significantly lower BChE activity, respectively, in the plasma of farmers exposed to pesticides, compared to the control group (p < 0.05). The measured mean level of BChE activity was (7304.80 ± 1939.99 U/L) and (9746.42 ± 1699.85 U/L) in the farmers and the control group (non-farmers), respectively. In addition, a high proportion of farmers reported that empty containers are burned in the open (74.6%) for waste disposal. A proportion (11.3%) of farmers also reported that empty container waste is spilled on the farm.Conclusions: The decrease in BChE indicates a serious public health problem among farmers who use organophosphate pesticides. This study suggests that regular monitoring for blood cholinesterase and effective interventions to reduce pesticide exposure to prevent health effects should be provided to farmers.


Assuntos
Biomarcadores/sangue , Butirilcolinesterase/sangue , Fazendeiros , Exposição Ocupacional/efeitos adversos , Adulto , Estudos de Casos e Controles , Inibidores da Colinesterase/efeitos adversos , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Marrocos , Organofosfatos/efeitos adversos , Praguicidas/efeitos adversos , Estudos Prospectivos
4.
Int J Biol Macromol ; 140: 1147-1157, 2019 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-31442505

RESUMO

In Alzheimer's disease (AD) and diabetes-associated cognitive decline, the acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) activity is increased. AChE exists as different globular molecular forms: tetramer (G4), dimer (G2) and monomer (G1). In adult brain, G4 form is abundant however in AD, the ratio of lower molecular forms (G1) to G4 form increased. Hence, the present study delineated the inhibition of novel astaxanthin-s-allyl cysteine (AST-SAC) against BChE and various molecular forms of AChE. Cobra venom, human erythrocyte and Electrophorus electricus was used as source of G1, G2 and G4 form of AChE. AST-SAC showed inhibition against G1 (IC50 = 0.72 µM, competitive, Ki = 0.66 µM), G2 (IC50 = 0.65 µM, mixed, Ki = 0.50 µM) and G4 (IC50 = 0.67 µM, competitive, Ki = 0.67 µM) form of AChE. AST-SAC inhibited human brain AChE (IC50 = 0.84 µM, competitive, Ki = 0.53 µM) and human serum BChE (IC50 = 0.80 µM, competitive, Ki = 0.58 µM). In silico analysis revealed the interaction of AST-SAC with the amino acids present in peripheral anionic and catalytic site of human AChE and BChE. Molecular dynamics simulation confirmed the stable interaction of AST-SAC in the active site gorge of AChE and BChE.


Assuntos
Acetilcolinesterase/metabolismo , Butirilcolinesterase/metabolismo , Inibidores da Colinesterase/farmacologia , Cisteína/análogos & derivados , Acetilcolinesterase/química , Animais , Encéfalo/enzimologia , Butirilcolinesterase/sangue , Butirilcolinesterase/química , Inibidores da Colinesterase/química , Simulação por Computador , Cisteína/química , Cisteína/farmacologia , Humanos , Simulação de Dinâmica Molecular , Xantofilas/química , Xantofilas/farmacologia
5.
Exp Parasitol ; 205: 107735, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31381870

RESUMO

Two experiments were performed to determine whether oral administration of copper oxide capsules controlled helminthic infections in Lacaune sheep without acute collateral effects on animal health. In experiment 1, 48 multiparous lactating sheep (60.1 ±â€¯8.5 kg) were stratified according to initial number of eggs (Haemonchus contortus) per gram of feces (EPG) and were assigned randomly to 1 of two treatments (24 sheep/treatment): no oral administration (control) or oral administration of two copper capsules (treated; approximately 58 mg copper/kg body weight). Blood and fecal samples were collected on days 0, 15 and 45. Animals treated with copper capsules showed lower of EPG, eosinophils, acetylcholinesterase (AChE) in whole blood, and lower butyrylcholinesterase (BChE) activity in serum. Treated sheep had higher erythrocyte numbers, hemoglobin concentrations, hematocrit, and lymphocyte numbers. In experiment 2, 12 male lambs negative for helminths and coccidia were assigned randomly to one of two treatments (six lambs/treatment): control or treated (one copper capsule; approximately 58 mg copper/kg body weight); the experiment was designed to determine whether the results of experiment 1 were due to treatment or parasitism. Blood samples were collected on days 0, 5, 10 and 15 and fecal samples were collected on days 0, 7 and 15. Treated animals showed greater concentrations of lymphocytes; however, treatment had no effect on other hemogram variables, AChE and BChE activities, or levels of alanine aminotransferase, aspartate aminotransferase, gamma-glutamyl transferase, creatinine, urea, albumin, total protein, and reactive oxygen species. These data suggest that copper capsules in dairy sheep efficiently controlled H. contortus infections. Treatment was not harmful to lambs during the first 15 days, i.e. there were no signs of acute toxicity.


Assuntos
Cobre/administração & dosagem , Hemoncose/veterinária , Helmintíase Animal/tratamento farmacológico , Lactação , Doenças dos Ovinos/tratamento farmacológico , Acetilcolinesterase/sangue , Administração Oral , Animais , Butirilcolinesterase/sangue , Cápsulas , Cobre/uso terapêutico , Indústria de Laticínios , Resíduos de Drogas , Eosinófilos/efeitos dos fármacos , Contagem de Eritrócitos/veterinária , Fezes/parasitologia , Feminino , Hemoncose/tratamento farmacológico , Hemoncose/prevenção & controle , Helmintíase Animal/sangue , Helmintíase Animal/prevenção & controle , Hematócrito/veterinária , Hemoglobinas/análise , Contagem de Linfócitos/veterinária , Masculino , Leite/química , Contagem de Ovos de Parasitas/veterinária , Paridade , Distribuição Aleatória , Ovinos , Doenças dos Ovinos/sangue , Doenças dos Ovinos/parasitologia , Doenças dos Ovinos/prevenção & controle
6.
Res Vet Sci ; 125: 290-297, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31349186

RESUMO

Endotoxemia treatment options are still of interest due to high mortality and choline treatment is one of them because of its role in the cholinergic anti-inflammatory pathway. This study investigated serum choline and butyrylcholinesterase (BChE) responses, and their correlations with inflammatory, oxidative stress and tissue damage biomarkers, including paraoxanase-1 (PON1), and clinical signs in calves with endotoxemia and the effect of choline treatment in these responses. Healthy calves (n = 20) were divided equally into 4 groups: Control (0.9% NaCl, iv), Choline (C; 1 mg/kg/iv,once), Lipopolysaccharide (LPS; 2 µg/kg/iv,once) and LPS + C. Clinical and laboratory examinations were performed before and 0.5-48 h (hrs) after treatments. Following LPS administration, serum choline level increased at 0.5-24 h (P < .01), whereas serum BChE and PON1 level decreased at 48 h (P < .01) compared to their baselines. In LPS + C group, the increase in serum choline level was significantly higher (P < .01) than that of C and LPS groups. LPS did not decrease serum BChE levels significantly in calves treated with choline. Serum choline and BChE results correlated negatively with white blood cell count and positively (P < .001) with PON1 levels, oxidative stress index, inflammation and hepato-muscular injury markers. In conclusion serum choline and BChE may have a role in the pathophysiology of endotoxemia in calves. High serum choline concentration is associated with an improvement in response to LPS administration in calves treated with choline, probably by preventing the imbalances between oxidative stress and anti-oxidant capacity, preventing the serum BChE and PON1 decreases, and inhibition/attenuation of acute phase reaction and hepato-muscular injury in calves with endotoxemia.


Assuntos
Butirilcolinesterase/sangue , Doenças dos Bovinos/induzido quimicamente , Colina/sangue , Endotoxemia/veterinária , Lipopolissacarídeos/toxicidade , Reação de Fase Aguda/tratamento farmacológico , Administração Intravenosa , Animais , Biomarcadores/sangue , Butirilcolinesterase/metabolismo , Bovinos , Doenças dos Bovinos/sangue , Doenças dos Bovinos/tratamento farmacológico , Endotoxemia/tratamento farmacológico , Endotoxemia/fisiopatologia , Inflamação/tratamento farmacológico , Contagem de Leucócitos , Lipopolissacarídeos/administração & dosagem , Masculino , Estresse Oxidativo , Distribuição Aleatória
7.
Exp Parasitol ; 204: 107726, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31299264

RESUMO

The aims of this study were to evaluate if the use of copper oxide wire particles, isolated or in association with closantel, in lambs infected with Haemonchus contortus enhances the anthelmintic efficacy of closantel, as well as to evaluate the effects of treatment in hepatic energy metabolism, inflammatory markers and hematological and biochemical tests. The lambs were randomly divided into five groups (6 animals each), as follows: uninfected animals (Control); animals infected with H. contortus (HC); infected and treated with closantel (HC + CL); infected and treated with copper oxide wire particles (HC + Cu); and infected and treated with closantel plus copper oxide wire particles (HC + CL + Cu). The animals of infected groups were infected orally with H. contortus (5,000 L3 -larvae) and on day 14 post infection (p.i) the treatments were initiated. The egg per gram of feces (EPG), butyrylcholinesterase (BuChE), myeloperoxidase (MPO), adenylate kinase (AK) and pyruvate kinase (PK) activities and hematological and biochemical tests were evaluated. Treatments with copper oxide (isolated and associated) were able to reduce the EPG count on days 28, 35, 42 and 49 p.i when compared to HC group, while closantel was able to reduce EPG only from day 35 p.i. Moreover, treatment with closantel (isolated or associated) was able to prevent the inhibition of hepatic AK and PK activities caused by H. contortus infection, which may contribute to efficient intracellular energetic communication in order to maintain the balance between cellular ATP consumption and production. Butyrylcholinesterase and MPO activities were higher in infected lambs compared to uninfected, while treated groups showed lower enzymatic activity compared to the group HC. The use of all therapeutic protocols was able to reduce the EPG count. Based on these evidences, the use of copper oxide plus closantel may be considered an alternative to treat lambs infected by H. contortus.


Assuntos
Anti-Helmínticos/administração & dosagem , Cobre/administração & dosagem , Hemoncose/veterinária , Inflamação/veterinária , Salicilanilidas/administração & dosagem , Doenças dos Ovinos/tratamento farmacológico , Abomaso/metabolismo , Adenilato Quinase/metabolismo , Administração Oral , Animais , Anti-Helmínticos/farmacologia , Anti-Helmínticos/uso terapêutico , Análise Química do Sangue/veterinária , Butirilcolinesterase/sangue , Cápsulas , Metabolismo Energético/efeitos dos fármacos , Contagem de Eritrócitos/veterinária , Fezes/parasitologia , Hemoncose/complicações , Hemoncose/tratamento farmacológico , Hemoncose/metabolismo , Hematócrito/veterinária , Hemoglobinas/análise , Inflamação/tratamento farmacológico , Inflamação/prevenção & controle , Fígado/efeitos dos fármacos , Fígado/enzimologia , Fígado/metabolismo , Masculino , Contagem de Ovos de Parasitas/veterinária , Peroxidase/sangue , Piruvato Quinase/metabolismo , Distribuição Aleatória , Salicilanilidas/farmacologia , Salicilanilidas/uso terapêutico , Ovinos , Doenças dos Ovinos/metabolismo , Doenças dos Ovinos/prevenção & controle
8.
Clin Lab ; 65(7)2019 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-31307170

RESUMO

BACKGROUND: Primary neonatal hypocholinesterase is rare; its genetic pattern and mutation still need to be further studied. METHODS: The patient and his parents are studied using next-generation sequencing technology. RESULTS: A boy one day after birth is admitted to the Neonatal Intensive Care Unit at our hospital after experiencing intermittent vomiting for 12 hours. The patient's serum cholinesterase level (113 - 283 U/L) is lower than normal value (4,000 - 12,600 U/L). Many factors of low serum cholinesterase are excluded. We highly suspect that it may be related to congenital factors. Molecular genetic test results show that the patient carried the BCHE gene (NM_000055.2) and has homozygous frameshift mutations at exon 2 c.401dupA (p.Asn134fs) of chromosome 3q26. It is a pathogenicity mutation. This locus mutation belongs to a novel pathogenic mutation. As a result of this mutation, the 134th amino acid Asn began to frameshift and the translation is terminated early. It can cause the Encoding of protein to truncate and lose its normal function. His parents' serum cholinesterase levels (father: 5,135 U/L; mother: 4,367 U/L) are in the normal value range, but his parents carried a heterozygous BCHE gene. CONCLUSIONS: This study suggests that gene sequence detection should be carried out early in hypocholinesterase of nknown cause in neonates. This study can not only improve understanding of the etiology and pathological mechanism of hypocholinesterase, but also it can enlarge the hypocholinesterase gene mutation spectrum.


Assuntos
Butirilcolinesterase/genética , Mutação da Fase de Leitura , Predisposição Genética para Doença/genética , Erros Inatos do Metabolismo/genética , Butirilcolinesterase/sangue , Butirilcolinesterase/deficiência , Saúde da Família , Sequenciamento de Nucleotídeos em Larga Escala , Homozigoto , Humanos , Recém-Nascido , Masculino , Erros Inatos do Metabolismo/sangue , Erros Inatos do Metabolismo/enzimologia
9.
Intern Med ; 58(20): 3057-3059, 2019 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-31243240

RESUMO

An 86-year-old woman intentionally drank approximately 300 mL of a glyphosate-surfactant. She was found with consciousness disturbance and experienced several vomiting episodes. On arrival, serum biochemistry revealed a decreased level of butyrylcholinesterase (B-CHE) [11 (normal range: 180-450) IU/L]. Later, her B-CHE level further decreased to single-digit values, and she became comatose with involuntary movement and an increase in muscle tone. Her consciousness level and muscle tone improved with the recovery of her B-CHE level. Physicians should be alert for the occurrence of intermediate syndrome when the B-CHE levels of patients who have consumed a massive amount of glyphosate-surfactant show a prolonged decrease.


Assuntos
Butirilcolinesterase/sangue , Coma/induzido quimicamente , Glicina/análogos & derivados , Tensoativos/efeitos adversos , Idoso de 80 Anos ou mais , Biomarcadores/sangue , Coma/sangue , Ingestão de Alimentos , Feminino , Glicina/envenenamento , Humanos , Síndrome
10.
Pestic Biochem Physiol ; 157: 138-142, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31153461

RESUMO

The evidence shows that organophosphate compounds (OPCs), as toxic agents that stimulate the cholinergic system, can increase the incidence of metabolic disorders such as dyslipidemia. In the present study, we focused on the role of tumor necrosis factor alpha (TNF-α) and serum leptin and ghrelin in Diazinon (DZN)-induced dyslipidemia. The rats were randomly divided into five groups comprising eight animals, and all were treated via oral gavage for 28 consecutive days as follows: group one received only corn oil daily, while groups two through five received different doses of DZN dissolved in corn oil equal to 1/40, 1/20, 1/10 and 1/5 of the LD50 daily, respectively. The alteration of the serum lipid profile, such as triglycerides, high-density lipoprotein (HDL) and very-low-density lipoprotein (VLDL), was confirmed the occurrence of dyslipidemia in the range of doses 1/20-1/5 LD50 of DZN. Although no changes were found in the serum leptin levels, a significant increase was observed in the size of adipocytes, as well as in the TNF-α and ghrelin serum levels, and in the accumulation of epididymal fat, especially at a dose of 1/5 LD50 of DZN. It seems that interactions among the inflammatory reaction, cholinergic pathways and ghrelin secretion may be effective causes of DZN-induced dyslipidemia.


Assuntos
Diazinon/farmacologia , Dislipidemias/sangue , Dislipidemias/induzido quimicamente , Grelina/sangue , Fator de Necrose Tumoral alfa/sangue , Animais , Antioxidantes , Butirilcolinesterase/sangue , Leptina/sangue , Peroxidação de Lipídeos/efeitos dos fármacos , Peroxidação de Lipídeos/fisiologia , Masculino , Compostos Organofosforados/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/fisiologia , Ratos
11.
Gac. sanit. (Barc., Ed. impr.) ; 33(3): 216-221, mayo-jun. 2019. tab
Artigo em Espanhol | IBECS | ID: ibc-183739

RESUMO

Objetivo: Evaluar el nivel de exposición a plaguicidas y su correlación con indicadores de salud percibida y biomarcadores de daño (alteraciones genotóxicas y de actividad enzimática de la butirilcolinesterasa), en la población de agroaplicadores de cultivos extensivos (ACE) de la Provincia de Córdoba, Argentina. Método: Estudio transversal, en ACE (n = 47) seleccionados aleatoriamente de una muestra de 2000, y sujetos no expuestos controles (n = 52). Se relevaron variables sociodemográficas, condicionantes de exposición y de salud percibida, mediante cuestionario autoadministrado; indicadores biológicos de genotoxicidad: micronúcleos, aberraciones cromosómicas y ensayo cometa, y actividad de butirilcolinesterasa. Resultados: El 40% de los ACE tiene una antigüedad mayor de 10 años y casi el 50% reside a menos de 500 m de campos asperjados; reportan bajas tasas de uso de equipo de protección personal durante la mezcla, aplicación o reparación de equipos. Los síntomas generales, cardiorrespiratorios y dermatológicos fueron mayores entre los ACE (p <0,05), así como los indicadores de daño genotóxico (p <0,001). La actividad butirilcolinesterasa se asoció negativamente a niveles de exposición a plaguicidas. Conclusiones: Los ACE presentan un importante impacto negativo en la salud vinculado a la exposición a plaguicidas. Las escalas de exposición asociadas al uso de biomarcadores resultaron una herramienta útil para la vigilancia de la salud de los agroaplicadores


Objective: To assess the level of exposure to pesticides and its correlation with perceived health indicators and injury biomarkers (genotoxic alterations and those caused by butyrylcholinesterase enzyme activity) in the population of pesticide applicators in extensive crops (PAEC) in Córdoba, Argentina. Methods: Transversal study, in PAEC (n = 47) randomly selected from a sample of 2000, and non-exposed subject controls (n = 52). The sociodemographic variables, exposure conditioning, and perceived health were surveyed by means of a self-administered questionnaire; biological indicators of genotoxicity: micronuclei, chromosomal aberrations and kite assay, and butyrylcholinesterase activity. Results: 40% of PAEC have over 10 years' length of service and almost 50% of them reside less than 500 m from the sprinkled fields; they report low rates of personal protective equipment use while mixing, applying, or repairing the equipment. General, cardio-respiratory, and dermatological symptoms were greater among PAEC (p <0.05) as well as indicators of genotoxic injury (p <0.001). The butyrylcholinesterase activity was negatively associated with levels of exposure to pesticides. Conclusions: The PAEC show an important negative impact on health linked to exposure to pesticides. The exposure scales associated to the use of biomarkers were a useful tool for monitoring pesticide applicators' health


Assuntos
Humanos , Masculino , Adulto , Pessoa de Meia-Idade , Monitoramento Ambiental/métodos , Exposição Ambiental/análise , Praguicidas/isolamento & purificação , Genotoxicidade/análise , Butirilcolinesterase/sangue , Monitoramento Epidemiológico , Argentina/epidemiologia , Estudos Transversais , Biomarcadores/análise , Estudos de Casos e Controles
12.
Chem Biol Interact ; 308: 170-178, 2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-31129133

RESUMO

Organophosphorus compounds (OP) pose a significant threat. Administration of human butyrylcholinesterase (HuBChE) may reduce or prevent OP toxicity. Thus, we evaluated the safety and efficacy of HuBChE in monkeys using sensitive neurobehavioral tests while concurrently characterizing absorption and elimination in the presence and absence of high-dose soman exposure to predict time course and degree of protection. Eight young adult male cynomolgus macaques were trained on two distinct automated tests of neurobehavioral functioning. HuBChE purified under current Good Manufacturing Practices (CGMP) was injected intramuscularly at 13.1 mg/kg, producing an average peak plasma value (Cmax) of over 27 Units/ml. The apparent time to maximum concentration (Tmax) approximated 7 h, the elimination half-life approximated 102 h, and plasma levels returned to pre-administration (baseline) levels by 14 days. No behavioral disruptions following HuBChE administration were observed on either neurobehavioral test, even in monkeys injected 24 h later with an otherwise lethal dose of soman. Thus, HuBChE provided complete neurobehavioral protection from soman challenge. The present data replicate and extend previous results from our laboratory that had used a different route of administration (intravenous), a different species (rhesus macaque), and a different BChE product (non-CGMP material). The addition of two sensitive neurobehavioral tests coupled with the PK/PD results convincingly demonstrates the neurobehavioral safety of plasma-derived HuBChE at therapeutic levels. Protection against an otherwise-lethal dose of soman by a pre-exposure treatment dose that is devoid of side effects establishes a foundation for additional testing using other exposure routes and treatment times, other challenge agents/routes, or other classes of organophosphate scavengers.


Assuntos
Comportamento Animal/efeitos dos fármacos , Butirilcolinesterase/administração & dosagem , Substâncias para a Guerra Química/metabolismo , Soman/metabolismo , Animais , Butirilcolinesterase/sangue , Butirilcolinesterase/farmacocinética , Substâncias para a Guerra Química/toxicidade , Meia-Vida , Humanos , Injeções Intramusculares , Macaca fascicularis , Masculino , Soman/toxicidade
13.
Life Sci ; 228: 98-111, 2019 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-31051153

RESUMO

AIMS: Protective efficacy of N­acetylcysteine (NAC) was assessed against sub-acute diisopropyl phosphorofluoridate (DFP) poisoning in mice. MAIN METHODS: Mice were allocated into nine groups of six each: vehicle control; DFP (0.125 LD50 ≈ 0.483 mg/kg bwt, s.c.); DFP + Atropine (ATR, 10 mg/kg bwt, i.p., 0 min); DFP + Pralidoxime (2-PAM, 30 mg/kg bwt, i.m., 0 min); DFP + NAC (150 mg/kg bwt, i.p., -60 min); DFP + ATR + NAC; DFP + 2-PAM + NAC; DFP + ATR + 2-PAM; and DFP + ATR + 2-PAM + NAC. Animals received various treatments for 21 d daily. Plasma butyrylcholinesterase (BChE) was measured after 7, 14 and 21 d of exposure. Brain acetylcholinesterase (AChE) and reduced glutathione (GSH), malondialdehyde (MDA), glutathione peroxidase (GPx), glutathione reductase (GR), catalase (CAT), and superoxide dismutase (SOD) were measured (brain, liver and kidney) after 21 d of exposure. Histopathology, immunohistochemistry, and Western blot for inducible nitric oxide synthase (iNOS) and c-fos were also performed. KEY FINDINGS: DFP significantly reduced BChE and AChE levels. Diminished GSH, CAT, SOD (brain and liver), GPx, GR, and elevated MDA (Brain) levels were also observed. DFP caused notable histopathology (brain, liver and kidney) and over expression of iNOS, and c-fos proteins (brain). NAC enhanced the protective efficacy of ATR and 2-PAM in most parameters, without any appreciable protection in iNOS and c-fos expression. SIGNIFICANCE: NAC as an adjunct with ATR and 2-PAM, exhibited marked beneficial effects against sub-acute DFP poisoning, indicating its possible implications in the management of OP poisoning.


Assuntos
Acetilcisteína/farmacologia , Antioxidantes/farmacologia , Encéfalo/efeitos dos fármacos , Inibidores da Colinesterase/toxicidade , Isoflurofato/toxicidade , Rim/efeitos dos fármacos , Fígado/efeitos dos fármacos , Acetilcolinesterase/análise , Animais , Encéfalo/patologia , Butirilcolinesterase/sangue , Catalase/análise , Glutationa/análise , Glutationa Peroxidase/análise , Glutationa Redutase/análise , Rim/patologia , Fígado/patologia , Masculino , Malondialdeído/análise , Camundongos , Estresse Oxidativo/efeitos dos fármacos , Superóxido Dismutase/análise
14.
Cardiovasc Toxicol ; 19(6): 548-564, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31098944

RESUMO

Previous studies showed that chlorpyrifos (CPF) acute exposure impaired cardiorespiratory reflexes. Evidence also indicates that continuous exposure to organophosphorus compounds impairs cardiovascular function. However, the effect of intermittent exposure to CPF, as may be experienced in the real world, on tonic and reflex cardiorespiratory function remains unexplored. Wistar rats were injected with saline or CPF for 4 weeks (3 times/week) or 12 weeks (once/week) at the doses of 7 mg/kg and 10 mg/kg. After exposure, blood pressure (BP), heart rate (HR), respiratory rate (fR), tidal volume (VT), and minute volume (VE) were recorded. Systolic BP and pulse interval (PI) variability, HR spectrum, spontaneous baroreflex and chemoreflex function were also evaluated. Plasma butyrylcholinesterase and brainstem acetylcholinesterase activities were quantified. Enzymatic activity of the CPF animals was reduced after both treatment periods. Baseline BP, HR, and fR, as well as systolic BP and PI variability indices, did not change, after CPF treatment. VT and VE were elevated in CPF animals. CPF exposure increased the very low-frequency component of the HR spectrum. Baroreflex gain was reduced after CPF 4-week exposure. Chemoreflex bradycardia was reduced in the CPF-treated rats. These data show that intermittent exposure to CPF impairs cardiorespiratory function in rats. These results may have important clinical implications for workers seasonally exposed to these compounds.


Assuntos
Barorreflexo/efeitos dos fármacos , Tronco Encefálico/efeitos dos fármacos , Clorpirifos/toxicidade , Inibidores da Colinesterase/toxicidade , Coração/inervação , Inseticidas/toxicidade , Pulmão/inervação , Acetilcolinesterase/metabolismo , Animais , Pressão Sanguínea/efeitos dos fármacos , Tronco Encefálico/enzimologia , Tronco Encefálico/fisiopatologia , Butirilcolinesterase/sangue , Cardiotoxicidade , Células Quimiorreceptoras/efeitos dos fármacos , Células Quimiorreceptoras/metabolismo , Proteínas Ligadas por GPI/antagonistas & inibidores , Proteínas Ligadas por GPI/metabolismo , Frequência Cardíaca/efeitos dos fármacos , Masculino , Ratos Wistar , Taxa Respiratória/efeitos dos fármacos , Volume de Ventilação Pulmonar/efeitos dos fármacos , Fatores de Tempo
15.
Toxicol Appl Pharmacol ; 372: 40-46, 2019 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-30978400

RESUMO

Tabun represents the phosphoramidate class of organophosphates that are covalent inhibitors of acetylcholinesterase (AChE), an essential enzyme in neurotransmission. Currently used therapy in counteracting excessive cholinergic stimulation consists of a muscarinic antagonist (atropine) and an oxime reactivator of inhibited AChE, but the classical oximes are particularly ineffective in counteracting tabun exposure. In a recent publication (Kovarik et al., 2019), we showed that several oximes prepared by the Huisgen 1,3 dipolar cycloaddition and related precursors efficiently reactivate the tabun-AChE conjugate. Herein, we pursue the antidotal question further and examine a series of lead precursor molecules, along with triazole compounds, as reactivators of two AChE mutant enzymes. Such studies should reveal structural subtleties that reside within the architecture of the active center gorge of AChE and uncover intimate mechanisms of reactivation of alkylphosphate conjugates of AChE. The designated mutations appear to minimize steric constraints of the reactivating oximes within the impacted active center gorge. Indeed, after initial screening of the triazole oxime library and its precursors for the reactivation efficacy on Y337A and Y337A/F338A human AChE mutants, we found potentially active oxime-mutant enzyme pairs capable of degrading tabun in cycles of inhibition and reactivation. Surprisingly, the most sensitive ex vivo reactivation of mutant AChEs occurred with the alkylpyridinium aldoximes. Hence, although the use of mutant enzyme bio-scavengers in humans may be limited in practicality, bioscavenging and efficient neutralization of tabun itself or phosphoramidate mixtures of organophosphates might be achieved efficiently in vitro or ex vivo with these mutant AChE combinations.


Assuntos
Antídotos/farmacologia , Butirilcolinesterase/metabolismo , Inibidores da Colinesterase/toxicidade , Reativadores da Colinesterase/farmacologia , Organofosfatos/toxicidade , Oximas/farmacologia , Acetilcolinesterase/química , Acetilcolinesterase/metabolismo , Antídotos/química , Butirilcolinesterase/sangue , Butirilcolinesterase/química , Domínio Catalítico , Reativadores da Colinesterase/química , Feminino , Proteínas Ligadas por GPI/antagonistas & inibidores , Proteínas Ligadas por GPI/química , Proteínas Ligadas por GPI/metabolismo , Humanos , Mutação , Oximas/química , Conformação Proteica , Proteínas Recombinantes/metabolismo , Relação Estrutura-Atividade
16.
Biosens Bioelectron ; 134: 97-102, 2019 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-30959394

RESUMO

In the last decades, there is a growing search for analytical strategies to ensure clinical analysis without the need of laboratory set-up and skilled personnel. Indeed, user-friendly and low-cost devices are highly valued in the era of sustainability for their capability to be applied in low-resource contexts, such as developing countries. To address this issue, herein we report a 96-well paper-based and laboratory setup-free optical platform for the detection of butyrylcholinesterase enzyme (BChE) activity in human serum. We used chromatographic paper to realize a novel analytical tool exploiting its porous structure for reagentless synthesize Prussian Blue Nanoparticles (the sensing element), as well to load all the reagents required for the measurement. The principle of BChE activity detection relies on the reaction between the enzymatic product thiocholine and Prussian Blue, giving the Prussian White with subsequently Prussian Blue's fading, detected by a common office scanner supported by ImageJ software. Using this novel paper-based optical platform, BChE activity was linearly detected in the 2-15 U/mL range with a detection limit down to 0.8 U/mL. The accuracy was successfully demonstrated by recovery study with spiked serum and by comparing the data with the gold standard method.


Assuntos
Técnicas Biossensoriais/instrumentação , Butirilcolinesterase/sangue , Colorimetria/instrumentação , Corantes/química , Ferrocianetos/química , Papel , Ceras/química , Desenho de Equipamento , Humanos , Limite de Detecção , Impressão Tridimensional
17.
Microb Pathog ; 132: 137-140, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31028864

RESUMO

The role of cholinesterase in inflammatory reactions has been described in several infectious diseases. However, in Brucella spp. this has not yet been studied. Therefore, the objective of this study was to evaluate whether experimental infection by Brucella ovis alters the cholinergic activity in pro- or anti-inflammatory responses to the disease. For the study 48 mice were used, 24 infected by B. ovis and 24 non-infected. We collected samples of whole blood on days 7, 15, 30 and 60 post-infection (PI) by B. ovis. Acetylcholinesterase (AChE) activity in the blood increased on days 15 and 60 PI (P < 0.05). Butyrylcholinesterase (BChE) activity in serum increased on days 7 and 60 PI (P < 0.05). An increase in serum free radical levels occurred on days 7, 15 and 60 PI (P < 0.05), and consequently superoxide dismutase activity increased on day 15 PI (P < 0.05). A reduction in catalase activity occurred when the infection became chronic (60 PI). The increase in AChE and BChE characterized a pro-inflammatory response, since these enzymes regulate levels of acetylcholine (ACh) and butyrylcholine (BuSCh), molecules with anti-inflammatory properties. Therefore, with the increase of cholinesterase activity, there was an extracellular reduction of ACh, an inhibitor of several inflammatory mediators. This proinflammatory response of B. ovis infection leads to oxidative stress, and consequently to cellular damage.


Assuntos
Acetilcolinesterase/metabolismo , Brucella ovis/patogenicidade , Butirilcolinesterase/metabolismo , Colinesterases/metabolismo , Acetilcolina/metabolismo , Acetilcolinesterase/sangue , Animais , Brucelose/sangue , Butirilcolinesterase/sangue , Catalase , Colina/análogos & derivados , Colina/metabolismo , Colinérgicos/farmacologia , Colinesterases/sangue , DNA Bacteriano/análise , Modelos Animais de Doenças , Inflamação , Masculino , Camundongos , Estresse Oxidativo , Espécies Reativas de Oxigênio/sangue , Espécies Reativas de Oxigênio/metabolismo , Soro/enzimologia , Superóxido Dismutase
18.
Parasitol Int ; 71: 11-17, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30849474

RESUMO

Both oxidative stress and alterations in adenosinergic and cholinergic systems participate in initiation and progression of parasitic infectious diseases. Nevertheless, the involvement of these pathways during eimeriosis remains poorly understood. Therefore, the aim of this study was to evaluate the involvement of adenosinergic and cholinergic systems in regulation of inflammatory response and oxidative stress in chicken chicks experimentally infected with Eimeria spp. Two groups were formed for comparison at 3 time points (days 5, 10 and 15) of infection (PI): uninfected (control) and infected. Erythrocyte counts, hematocrit and hemoglobin levels were lower in infected chicks on day 15 post-infection (PI). Total leukocytes, heterophil and lymphocyte counts were higher in infected chicks on days 5 and 10 PI, while eosinophil counts were higher only on day 10 PI. Serum levels of total protein and globulins were higher in infected chicks on days 10 and 15 PI, while triglycerides and cholesterol levels were lower on day 15 PI. Acetylcholinesterase activity in total blood and butyrylcholinesterase activity in serum were higher in infected chicks on day 15 PI, while adenosine deaminase activity was higher on day 10 PI and lower on day 15 PI compared with the respective control. Finally, serum levels of reactive oxygen species and catalase activity in total blood were higher in infected chicks on day 15 PI, while superoxide dismutase activity in total blood was lower at the same time of infection. These data suggest that cholinergic and adenosinergic systems display a pro-inflammatory profile that contributes to impairment of immune and inflammatory responses in a mixed Eimeria infection. Furthermore, oxidative stress may contribute to clinical signs of disease as well as to pathogenesis. In summary, the impairment of immune response and alterations in blood antioxidant/oxidant status contributes to disease pathophysiology.


Assuntos
Acetilcolinesterase/sangue , Adenosina Desaminase/sangue , Butirilcolinesterase/sangue , Coccidiose/veterinária , Eimeria , Estresse Oxidativo , Animais , Catalase/sangue , Galinhas/parasitologia , Coccidiose/fisiopatologia , Inflamação , Oócitos/fisiologia , Doenças das Aves Domésticas/sangue , Doenças das Aves Domésticas/parasitologia , Espécies Reativas de Oxigênio/sangue
19.
Biosens Bioelectron ; 131: 232-236, 2019 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-30849722

RESUMO

A ratiometric fluorescence probe based on carbon dots (CDs) was developed for discriminative and highly sensitive detection of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) activity in human whole blood. When o-phenylenediamine (OPD) was oxidized by Cu2+, the product 2,3-diaminophenazine (oxOPD) could effectively quench the fluorescence of CDs at 460 nm due to the inner filter effect and gave rise to a new emission peak at 570 nm. The AChE or BChE catalyzed hydrolysis reaction of acetylthiocholine or butyrylthiocholine to generate thiocholine, whose sulfhydryl group strongly captured Cu2+ to inhibit the oxidization of OPD, thus effectively preserving the natural fluorescence emission of CDs. The resulting fluorescence intensity ratio served as the signal output of the probe for cholinesterases (ChEs) activity sensing. The activities of AChE and BChE were determined to range from 0.2 to 14.0 U L-1 and from 0.1 to 5.0 U L-1, with detection limits of 0.1 U L-1 and 0.04 U L-1, respectively. Additionally, the IC50 of tacrine and ethopropazine for the inhibition of AChE and BChE were estimated to be 29.8 nM and 132.6 nM, respectively. Moreover, the probe was successfully applied to the discriminative determination of AChE and BChE in human whole blood without any pretreatment. These results suggested that the proposed strategy provided a discriminative, sensitive and robust analytical platform for ChEs clinical diagnostics and drug screening.


Assuntos
Acetilcolinesterase/sangue , Técnicas Biossensoriais , Butirilcolinesterase/sangue , Carbono/química , Catálise , Inibidores da Colinesterase/química , Cobre/química , Corantes Fluorescentes/química , Humanos , Limite de Detecção , Fenazinas/química , Fenilenodiaminas/química
20.
Ecotoxicol Environ Saf ; 172: 246-254, 2019 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-30711859

RESUMO

Free-ranging animals are often used as bioindicators of both short- and long-term changes in ecosystem health, mainly to detect the presence and effects of contaminants. Birds, and gulls in particular, have been used as bioindicators over a broad range of marine and terrestrial ecosystems. In this study, we standardise the conditions for the use of a suite of biochemical markers in non-destructive matrices of Lesser Black-backed Gull (Larus fuscus) to facilitate future biomonitoring of marine and terrestrial contaminants. We characterized cholinesterase (ChE) in plasma and optimized assay conditions for ChE activity as a marker of neurotoxic damage. Moreover, we quantified variation in activity of ChE, lactate dehydrogenase (LDH), glutathione-S-transferase (GST) and catalase (CAT) as well as variation ranges of lipid peroxidation (LPO), in free-ranging adults and captive chicks. The main ChE form present in plasma of both adults and chicks was butyrylcholinesterase (BChE) followed by acetylcholinesterase (AChE), whose relative proportion in plasma tended to decrease with increased chick age. LPO levels and GST activity in blood cells (BCs) decreased significantly with increasing chick age, while BChE and LDH activity in plasma were not age-dependent. CAT in BCs tended to decline non-significantly in older chicks. Results of this study underscore the importance of standardising assay conditions and assessing intrinsic baseline variation in biochemical markers, before biochemical quantification. Data presented here provide a foundation for future use of BChE and LDH activity in plasma, as well as oxidative stress markers (LPO, CAT and GST) in BCs, to monitor environmental stress effects in Lesser Black-backed gulls.


Assuntos
Charadriiformes/metabolismo , Monitoramento Ambiental/métodos , Acetilcolinesterase/sangue , Fatores Etários , Animais , Bioensaio , Biomarcadores/metabolismo , Butirilcolinesterase/sangue , Catalase/metabolismo , Feminino , Glutationa Transferase/metabolismo , L-Lactato Desidrogenase/metabolismo , Peroxidação de Lipídeos , Masculino
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