Synthesis and Biological Activity of Trolox Amide Derivatives

Abstract A series of Trolox amide derivatives were synthesized by modifying the carboxyl groups of Trolox. Thirty target compounds were obtained and characterized through nuclear magnetic resonance and mass spectrometry. Trolox derivatives were employed to explore the potential structure-antioxidant activity relationships. The antioxidant activities of these compounds were evaluated using 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), ferric reducing antioxidant power (FRAP) and hydroxyl radical assays. DPPH scavenging activity test results illustrated that compounds exhibited scavenging activities similar to L-ascorbic acid and Trolox, with compounds 14a, 18a, 24a and 26a in particular exhibiting higher scavenging activities than L-ascorbic acid. The results demonstrated that compounds displayed ABTS scavenging activities similar to L-ascorbic acid and Trolox, with compounds 26a and 29a in particular having potency twofold higher. FRAP assay results indicated that compounds 11a, 19a, 25a, 29a and 30a had activity similar to Trolox. The results revealed that compounds 6a and 19a had similarly high hydroxyl radical-scavenging activities as Trolox. The results of α-glucosidase experiments uncovered that compounds 10a, 25a, 28a and 29a had excellent inhibitory activity, which was similar to that of acarbose and different from Trolox. The results of acetylcholinesterase and butyrylcholinesterase experiments demonstrated that some compounds had weak anticholinesterase activities. 26a and 29a are important Trolox derivatives with better biological activity profiles and deserve further study

Chemical profile, acetylcholinesterase, butyrylcholinesterase, and prolyl oligopeptidase inhibitory activity of Glaucium corniculatum subsp. refractum

Abstract Papaveraceae is one of the prominent alkaloid-containing families, and plants of the genus Glaucium (Papaveraceae) are known for their bioactive alkaloids. Glaucium species have been used in traditional medicine in Turkey as an analgesic, narcotic, sedative, and antitussive. In this study, it was planned to evaluate the inhibitory activity of an alkaloidal extract of Glaucium corniculatum subsp. refractum on acetylcholinesterase (AChE), butyrylcholinesterase (BuChE) and prolyl oligopeptidase (POP), as well as exploring the chemical profile of the plant by using Gas Chromatography-Mass Spectrometry (GC-MS). The AChE, BuChE and POP inhibition activities of the alkaloidal extract of G. corniculatum subsp. refractum were determined spectrophotometrically. A rapid GC-MS method was used to identify alkaloids that could be responsible for these inhibition activities. In total, eleven alkaloids were identified in the alkaloid extract of the plant by GC-MS. Allocyptopine (52.92%) and protopine (25.38%) were found as the major constituents. The alkaloidal extract of G. corniculatum subsp. refractum showed potent AChE inhibitory activity (IC50:1.25 µg/mL) and BuChE inhibitory activity (IC50: 7.02 µg/mL). The extract also showed a remarkable inhibitory effect on POP with an IC50 value of 123.69 µg/mL. This study presents the first GC-MS investigation and POP inhibitory activity of G. corniculatum subsp. refractum.

Crude extract of the tropical tree Gallesia integrifolia (Phytolaccaceae) for the control of Aedes aegypti (Diptera: Culicidae) larvae / Extracto crudo del árbol tropical Gallesia integrifolia (Phytolaccaceae) para el control de larvas de Aedes aegypti (Diptera: Culicidae)

Introduction: Phytoinsecticides are alternatives to control insects in different stages, Gallesia integrifolia (Spreng.) Harms, Phytolacaceae family, popularly known as pau d'alho, garlic tree, and guararema in Brazil, is known due to its strong alliaceous odor because of the presence of sulfur molecules in the plant. This species presents biological activity and potential insecticide effect that is still unexploited. Objective: This study aimed to evaluate the biological activity of the ethanolic crude extract from G. integrifolia leaves, flowers, and fruits on the control of Aedes aegypti third-stage larvae and pupae. Methods: The botanical material was collected in city Umuarama, Paraná, Brazil at the coordinates (23º46'16" S & 53º19'38" WO), and altitude of 442 m, the fruits of G. integrifolia were collected in May and the leaves and flowers in December 2017. The crude extracts of G. integrifolia leaves, flowers, and fruits were prepared by dynamic maceration technique. The chemical composition of the extracts was determined by gas chromatography coupled to a mass spectrometry. The insecticidal activity of the crude extracts of G. integrifolia were carried out on larvae and pupae of A. aegypti in concentrations between 0.001 to 25 000 mg/mL, and afterwards the lethal concentrations that kill 50 % (LC50) and 99.9 % (LC99.9) were determined by probit analysis. Anticholinesterase activity was determined by bioautographic method at concentrations from 0.000095 to 50 mg/mL. Results: The yield of G. integrifolia crude extracts were 8.2, 9.1, and 17.3 % for flowers, fruits, and leaves, respectively. The chemical composition of G. integrifolia extracts was characterized by presence of fatty acid esters, phytosterols, vitamins, oxygenated diterpenes and organosulfur compounds. The flower extract presented the high amount of sulfur compounds (20.2 %) such as disulfide, bis (2-sulfhydryl ethyl) (11.9 %), 2,3,5-trithiahexane (6.2 %), 1,2,4-trithiolane (1.1 %), and 2,4-dithiapentane (1.1 %). Regarding the insecticidal activity, flower extract showed highly active with LC99.9 of 0.032 mg/mL and LC99.9 of 0.969 mg/mL on A. aegypti larvae and pupae, respectively, and the highest inhibition of acetylcholinesterase enzyme (0.00019 mg/mL) ex situ. The flower extract presented anticholinesterase and larvicide activity, respectively, 12.8 % and 35.6 % greater than the control temephos. Conclusions: This study opens new perspectives on the use of extracts from G. integrifolia as a bioinsecticide alternative for the control of A. aegypti larvae and pupae.

Introducción: Los fitoinsecticidas son alternativas para el control de insectos en diferentes etapas, Gallesia integrifolia (Spreng.) Harms, familia Phytolacaceae, conocida popularmente como pau d'alho, árbol de ajo y guararema en Brasil, es conocida por su fuerte olor aliáceo debido a la presencia de moléculas de azufre en la planta. Esta especie presenta actividad biológica y potencial efecto insecticida que aún no está explotado. Objetivo: El objetivo de este estudio fue evaluar la actividad biológica del extracto crudo etanólico de las hojas, flores y frutos de G. integrifolia en el control de las larvas y pupas de la tercera etapa de Aedes aegypti. Métodos: El material botánico fue recolectado en la ciudad de Umuarama, Paraná, Brasil (23º46'16" S & 53º19'38" W), a una altitud de 442 m y los frutos de G. integrifolia fueron recolectados en mayo de 2017 y las hojas y flores en diciembre del mismo año. Los extractos crudos de hojas, flores y frutos de G. integrifolia se prepararon por técnica de maceración dinámica. La composición química de los extractos se determinó por cromatografía de gases acoplada a espectrometría de masas. La actividad insecticida de los extractos crudos de G. integrifolia fue evaluada en larvas y pupas de A. aegypti en concentraciones entre 0.001 a 25 000 mg/mL, y las concentraciones letales que matan 50 % (CL50) y 99.9 % (CL99.9) de larvas fueron determinados por análisis de probit. La actividad anticolinesterasa fue evaluada por método bioautográfico en concentraciones de 0.000095 a 50 mg/mL. Resultados: El rendimiento de los extractos crudos de G. integrifolia fue de 8.2, 9.1 y 17.3 % para flores, frutos y hojas, respectivamente. La composición química de los extractos de G. integrifolia se caracterizó por la presencia de ésteres de ácidos grasos, fitosteroles, vitaminas, diterpenos oxigenados y compuestos organosulfurados. El extracto de las flores presentó alta cantidad de compuestos de azufre (20.2 %) como disulfuro, bis(2-sulfhidril etilo) (11.9 %), 2,3,5-tritiahexano (6.2 %), 1,2,4-tritiolano (1.1 %) y 2,4-ditiapentano (1.1 %). En relación con la actividad insecticida, el extracto de las flores mostró una gran actividad con CL99.9 de 0.032 mg/mL y CL99.9 de 0.969 mg/mL en larvas y pupas de A. aegypti, respectivamente, y la inhibición más alta de la enzima acetilcolinesterasa (0.00019 mg/mL) ex situ. El extracto de las flores presentó actividad anticolinesterasa y larvicida, 12.8 y 35.6 %, respectivamente, mayor que el control temephos. Conclusiones: Este estudio abre nuevas perspectivas sobre el uso de extractos de G. integrifolia como alternativa bioinsecticida para el control de larvas y pupas de A. aegypti.

Insecticidal activity of Gallesia integrifolia (Phytolaccaceae) essential oil / Actividad insecticida del aceite esencial de Gallesia integrifolia (Phytolaccaceae)

This study evaluated the insecticidal activity of Gallesia integrifolia essential oil from fruits, leaves, and flowers against Aedes aegyptilarvae and pupae. The essential oil was obtained by hydrodistillation and characterized by gas chromatography-mass spectrometry. Sulfur compounds represented 95 to 99% of the essential oil from fruits, leaves, and flowers. Essential oil major compounds were 2,8-dithianonane (52.6%) in fruits, 3,5-dithiahexanol-5,5-dioxide (38.9%) in leaves, and methionine ethyl ester (45.3%) in flowers. The essential oils showed high activity against larvae, and low for pupae withLC99.9of 5.87 and 1476.67µg/mL from fruits; 0.0096 and 348.33 µg/mL from leaves and 0.021and 342.84 µg/mL from flowers, respectively. The main compound with insecticide activity is probably n-ethyl-1,3-dithioisoindole, from isoindole organothiophosphate class, found in greater amount in flower and leaf essential oil. The great insecticide activity of G. integrifolia essential oil suggests that this product is a natural insecticide.

Este estudio evaluó la actividad insecticida del aceite esencial de frutos, hojas y flores de Gallesia integrifolia contra larvas y pupas de Aedes aegypti. El aceite esencial se obtuvo por hidrodestilación y se caracterizó por cromatografía de gases acoplada a espectrometría de masas. Los compuestos de azufre representaron del 95 al 99% del aceite esencial de frutas, hojas y flores. Los compuestos principales del aceite esencial fueron 2,8-ditianonano (52,6%) en frutas, 3,5-ditiahexanol-5,5-dióxido (38,9%) en hojas y éster etílico de metionina (45,3%) en flores. Los aceites esenciales mostraron alta actividad contra larvas y baja para pupas con LC99.9de 5.87 y 1476.67µg/mL de frutos; 0,0096 y 348,33 µg/mL de hojas y 0,021 y342,84 µg/mL de flores, respectivamente. El principal compuesto con actividad insecticida es probablemente el n-etil-1,3-ditioisoindol, de la clase de los organotiofosfatos de isoindol, que se encuentra en mayor cantidad en el aceite esencial de flores y hojas. La gran actividad insecticida del aceite esencial de G. integrifolia sugiere que este producto es un insecticida natural.

Anti-tick effect and cholinesterase inhibition caused by Prosopis juliflora alkaloids: in vitro and in silico studies / Efeito carrapaticida e inibição da acetilcolinesterase por alcaloides de Prosopis juliflora: estudos in vitro e in silico

Abstract We investigated the in vitro acaricide activity of the methanolic extract (ME) and alkaloid-rich fraction (AF) of Prosopis juliflora on Rhipicephalus microplus and correlated this effect with acetylcholinesterase (AChE) inhibition. The acaricide activity was evaluated using adult and larval immersion tests. Also, we studied the possible interaction mechanism of the major alkaloids present in this fraction via molecular docking at the active site of R. microplus AChE1 (RmAChE1). Higher reproductive inhibitory activity of the AF was recorded, with effective concentration (EC50) four times lower than that of the ME (31.6 versus 121 mg/mL). The AF caused mortality of tick larvae, with lethal concentration 50% (LC50) of 13.8 mg/mL. Both ME and AF were seen to have anticholinesterase activity on AChE of R. microplus larvae, while AF was more active with half-maximal inhibitory concentration (IC50) of 0.041 mg/mL. The LC-MS/MS analyses on the AF led to identification of three alkaloids: prosopine (1), juliprosinine (2) and juliprosopine (3). The molecular docking studies revealed that these alkaloids had interactions at the active site of the RmAChE1, mainly relating to hydrogen bonds and cation-pi interactions. We concluded that the alkaloids of P. juliflora showed acaricide activity on R. microplus and acted through an anticholinesterase mechanism.

Resumo A atividade carrapaticida in vitro do extrato metanólico (EM) e da fração de alcaloides (FA) de Prosopis juliflora foi investigada, frente ao Rhipicephalus microplus, e relacionada com a inibição da enzima acetilcolinesterase (AChE). A predição in silico das interações de alcaloides dessa fração com a AChE1 de R. microplus (RmAChE1) foi realizada por acoplamento molecular. A atividade carrapaticida foi avaliada, utilizando-se os ensaios de imersão de adultos e larvas. Maior efeito sobre parâmetros reprodutivos de teleóginas foi verificado para a FA, com valor de Concentração Efetiva 50% (CE50) (31.6 mg/mL), quatro vezes menor do que o valor do EM (121 mg/mL). A FA induziu mortalidade de larvas (Concentração Letal de 50% - CL50 = 13,8 mg/mL). A inibição da atividade da AChE de larvas do carrapato foi observada para EM e FA, sendo a FA mais ativa (Concentração Inibitória 50%- CI50 de 0,041mg/mL). As análises químicas da FA permitiram a identificação dos alcaloides prosopina (1), juliprosinina (2) e juliprosopina (3). No ensaio in silico, observou-se que esses alcaloides podem interagir com o sítio ativo da RmAChE1, principalmente por ligações de hidrogênio e interações cátion-pi. Os alcaloides de P. juliflora têm atividade carrapaticida contra R. microplus, atuando através do mecanismo anticolinesterásico.

Phenolic compounds profile, neuroprotective effect and antioxidant potential of a commercial Turkish coffee / Perfil do componente fenólico, efeito neuroprotector e potencial antioxidante de um café comercial turco

ABSTRACT Objective The purpose of this study is to determine the phenolic and flavonoid contents, and antioxidant activities and neuroprotective effects of powdered coffee sample of a commercial coffee brand originated from Sivas, Turkey. Methods Total phenolic, flavonoid and antioxidant contents, enzymatic and non-enzymatic antioxidative activities based on 2,2-diphenyl-1-picrylhydrazyl free radical scavenging activity, metal chelating potential, reducing power, superoxide dismutase and catalase activity tests and lipid peroxidation inhibition potentials of the ethanolic and aqueous extracts of the coffee sample were assayed using the commonly preferred spectrophotometric methods. Furthermore the extracts' cholinesterase and tyrosinase inhibition potentials were evaluated. Phenolic profiles of the coffee sample were investigated using high performance liquid chromatography. Results Catechin was the most frequently detected phenolic acid. In addition, it was demonstrated that the water extract has a significant impact when compared with standard antioxidants. While the SC50 (sufficient concentration to obtain 50% of a maximum scavenging capacity) value for the scavenging activity of 2,2-diphenyl-1-picrylhydrazyl free radical was calculated as being 0.08mg/mL for water extract, the amount of chelating agents with half Fe2+ ions in the medium was found to be 0.271mg/mL. Additionally, it was shown that 0.1mg/mL concentration of both extracts prevents lipid peroxidation by 8%. Compared with standard drugs, inhibition potentials of cholinesterase and tyrosinase enzymes were considered as moderately acceptable in these samples. Conclusion Besides the extracts' enzymatic antioxidant activity, their inhibition potential on cholinesterase and tyrosinase enzymes - which are important clinical enzymes - reveal that this natural source can be used as a valuable resource in different fields, especially in medicine.

RESUMO Objetivo O objetivo deste estudo é determinar o conteúdo fenólico e flavonoide, bem como as atividades antioxidantes e os efeitos neuroprotetores de uma amostra de café em pó de uma promissora marca comercial proveniente de Sivas, Turquia. Métodos A partir dos métodos espectrofotométricos comumente utilizados, foram analisados os seguintes aspectos da amostra de café: teores de fenólicos totais, flavonoides e antioxidantes; atividades antioxidantes enzimáticas e não enzimáticas, baseadas na atividade de eliminação de radicais livres de 2,2-difenil-1-picrilhidrazila potencial quelante de metais; poder redutor; testes de atividade de superóxido dismutase e catalase; e potenciais de inibição da peroxidação lipídica dos extratos etanólicos e aquosos. Além disso, foram avaliados os potenciais de inibição da colinesterase e da tirosinase dos extratos. Os perfis fenólicos da amostra de café foram investigados por cromatografia líquida de alta eficiência. Resultados Entre os ácidos fenólicos estudados, o mais detectado foi a catequina. Especialmente, foi demonstrado que o extrato de água tem um impacto significativo quando comparado com os antioxidantes padrão. Determinou--se que o valor de SC50 (a concentração suficiente para obter 50% da capacidade máxima de eliminação) da atividade de eliminação do radical 2,2-difenil-1-picrilhidrazilab/para extrato de água era de 0,08mg/mL, enquanto a quantidade de agentes quelantes com metade de Fe2+ íons na média foi encontrada como 0,271mg/mL. Também foi demonstrado que a concentração de 0,1mg/mL de ambos os extratos inibe a peroxidação lipídica em cerca de 8%. Comparado com drogas padrão, os potenciais de inibição das amostras nas enzimas e tirosinase foram aceitáveis como moderados. Conclusão Os resultados mostram que, além de terem atividade antioxidante enzimática, os extratos apresentam potencial de inibição das enzimas colinesterase e tirosinase, que são importantes enzimas clínicas, o que revela que essa fonte natural pode ser usada como um recurso valioso em vários campos, principalmente na medicina.

The in vitro and in vivo investigation of biological activities and phenolic analysis of Helichrysum plicatum subsp. plicatum

In Turkey, Helichrysum genus is represented by 26 taxa belonging to 20 species in Turkish flora of which 14 ones are endemic to Turkey. The aerial parts of Helichrysum plicatum subsp. plicatum are used kidney stones, kidney and stomach ailments. The extraction procedures and solvents are important step in processing of bioactive constituents from the plant materials. Therefore, the aim of this study was to evaluate in vitro antioxidant, antimicrobial, anti-urease, anticholinesterase and in vivo anti-inflammatory activities of Helichrysum plicatum subsp. plicatum different extracts. In addition, the phenolic characterization of the Soxhlet and maceration methanol extracts which showed significant antioxidant, anti-urease, antimicrobial, anti-inflammatory and anticholinesterase activities were performed by HPLC-DAD and LC-MS/MS. In the present study, the Soxhlet methanol extract exhibited strong antioxidant, antimicrobial and anticholinesterase activities than other extracts. The maceration methanol extract showed the strongest anti-urease activity. The Soxhlet methanol and maceration methanol extracts showed in vivo anti-inflammatory activities very close to each other. As a result of this study, chlorogenic acid, dicaffeoylquinic acid, luteolin, luteolin-7-O-glucoside, naringenin-O-hexoside and isoquercitrin compounds were analysed in plant. Therefore, it is thought that methanol extracts can be used as a natural source in the future for food and pharmaceutical industries.

Bioinsecticide potential of Curcuma zedoaria rhizome essential oil / Potencial bioinseticida do óleo essencial dos rizom as de curcuma zedoaria

In this study the potential bioinseticide of the essential oil (OE) extracted from the rhizomes of the species Curcuma zedoaria (Zingiberaceae) was evaluated. The rhizomes were collected during dormancy (winter) and budding (summer). The EO was obtained by hydrodistillation (2h) and identified by GC/MS. In addition, a multivariate exploratory analysis was done to determine the analysis of the major compounds (PCA). The EO yield in dormancy was 0.61± 0.07 (%) and in budding 0.55 ± 0.08 (%). The bioassays on Aedes aegypti larvae and pupae were done by immersion test at different EO concentrations which ranged from 500.00 to 0.003 mg mL-1 (v/v). The results on the larvae and pupae indicated LC99.9 of (0.01 and 1.38 mg mL-1) for EO in dormancy, and (0.08 and 2.63 mg mL-1) for EO during budding, respectively. The action mechanism of EOs in both periods was determined by autobiographic method evaluating the inhibitory potential on the acetylcholinesterase enzyme, indicating greater inhibition of the EO enzyme during dormancy (0.039 mg mL-1) when compared to the EO during budding (0.156 mg mL-1). The projection representation of the EO chemical classes in both evaluated periods indicated that oxygenated sesquiterpenes are the major compound class (46.99% in dormancy) and (43.59% in budding). The projection of major chemical compounds of EOs presented three compounds with greater mass flow distancing: epicurzerenone (18.20% and 12.10%); 1.8 cineole (15.76% and 12.10%) and ß-elemene (4.43 and 0.01%) that are found in greater amounts in the dormancy EO when compared to budding, respectively. These results corroborate with the greater potential on Ae. aegypti larvae and pupae found for the dormancy EO. The results are promising because they show in which vegetative cycle phase C. zedoaria EO presents greater bioinsecticidepotential.

Neste trabalho foi avaliado o potencial bioinseticida do óleo essencial (OE) extraído dos rizomas da espécie Curcuma zedoaria (Zingiberaceae), coletados no período de dormência (inverno) e brotação das gemas (verão). O OE foi obtido por hidrodestilação (2h) e identificado por CG/EM foi observado rendimento 0,61 ± 0,07 (%) no óleo da dormência, quando comparado no período de brotação 0,55 ± 0,08 (%). Os bioensaios sobre as larvas e pupas de Aedes aegypti foram realizados pelo teste de imersão em diferentes concentrações dos OEs, que variaram de 500,00 a 0,003 mg mL-1 (v/v). Os resultados sobre as larvas e pupas indicaram uma CL99,9 de (0,01 e 1,38 mg mL-1) para o OE da dormência, e (0,08 e 2,63 mg mL-1) para o OE do período de brotação, respectivamente. Indicando maior atividade do OE da dormência. O mecanismo de ação dos OEs nos dois períodos foi determinado pelo método autobiográfico avaliando o potencial inibitório sobre a enzima acetilcolinesterase. Os resultados indicaram maior inibição da enzima do OE no período de dormência (0,039 mg mL-1), quando comparado ao OE de brotação (0,156 mg mL-1). A análise química destacou três compostos: epicurzerenone (18,20% e 12,10%) e 1,8 cineol (15,76% e 14,05%) e ß- elemeno (4,43 e 0,01%) em maior quantidade no período de dormência quando comparado ao período de brotação, respectivamente. Esta diferença pode explicar a maior ação inseticida do OE de dormência sobre as larvas e pupas do Ae. aegypti. Os resultados são promissores, pois estabelece em qual período do ciclo vegetativo o OE da C. zedoaria apresenta maior potencial bioinseticida.

Biological screening of extracts from leaf and stem bark of Croton floribundus Spreng. (Euphorbiaceae) / Triagem biológica de extratos das folhas e caules de Croton floribundus Spreng. (Euphorbiaceae)

Abstract This work describes the preliminary evaluation of cytotoxic, antimicrobial, molluscicidal, antioxidant and anticholinesterase activities from leaf (LECF) and stem bark alcoholic extracts (BECF) of the species Croton floribundus Spreng. (Euphorbiaceae), popularly known as capixingui or tapixingui. BECF presented significant toxicity (LC50 = 89.6 μg/ml) in the Artemia salina Leach, 1819 (Crustacea: Branchiopoda) bioassay, whereas LECF did not show activity (LC50 > 1000 μg/ml). From DPPH method, the values of IC50 for the LECF and BECF were 61.2 μg/ml and 62.2 μg/ml, respectively, showing that C. floribundus has an expressive antioxidant activity. Antimicrobial susceptibility was evaluated by microdilution technique and only BECF was active against Staphylococcus aureus (MIC = 39.6 μg/ml). The extracts did not present molluscicidal activity against snail Biomphalaria glabrata Say, 1818 (Gastropoda: Planorbidae). Both extracts revealed the presence of several components with an inhibiting capacity of acetylcholinesterase enzyme on the bioautographic assay. C. floribundus showed to be a promising species considering that it exhibited good biological activity in the most assays performed.

Resumo Este trabalho descreve a avaliação preliminar das atividades citotóxica, antimicrobiana, moluscicida, antioxidante e anticolinesterásica de extratos alcoólicos das folhas (LECF) e das cascas do caule (BECF) da espécie Croton floribundus Spreng. (Euphorbiaceae), popularmente conhecida como capixingui ou tapixingui. No bioensaio com Artemia salina Leach, 1819 (Crustacea: Branchiopoda), BECF apresentou toxicidade significante (LC50 = 89,6 µg/ml), enquanto que LECF não apresentou atividade (LC50 > 1000 µg/ml). A partir do método de DPPH, os valores de IC50 para o LECF e BECF foram 61,2 µg/ml e 62,2 µg/ml, respectivamente, evidenciando que C. floribundus tem uma atividade antioxidante expressiva. A susceptibilidade antimicrobiana foi avaliada pela técnica de microdiluição e apenas BECF foi ativo contra Staphylococcus aureus (MIC = 39,6 mg/ml). Os extratos não apresentaram atividade moluscicida contra o caramujo Biomphalaria glabrata Say, 1818 (Gastropoda: Planorbidae). Ambos os extratos revelaram a presença de componentes com capacidade inibidora da enzima acetilcolinesterase no ensaio bioautográfico. C. floribundus mostrou ser uma espécie promissora considerando que exibiu boa atividade biológica na maioria dos ensaios testados.

Carnosol from Lepechinia mutica and tiliroside from Vallea stipularis: Two promising inhibitors of BuChE

Abstract Lepechinia mutica (Benth.) Epling, Lamiaceae, and Vallea stipularis L.f., Elaeocarpaceae, are the object of the present study. These plants are endemic to the Andean region and have attracted our attention on the basis of interesting results obtained in a preliminary anticholinesterase screening. Actually, carnosol and tiliroside, isolated from L. mutica and V. stipularis, respectively, have shown a promising selective inhibitory activity against butyrylcholinesterase. Specifically, the anti-butyrylcholinesterase activity of carnosol was 5.15 µM and that of tiliroside was 52.9 µM, compared to 8.568 ± 0.570 µM of the positive control Donepezil. Carnosol and tiliroside were purified chromatographically from the ethyl acetate extract of L. mutica and V. stipularis, respectively. Spectrophotometric methods were used for enzymatic studies.

Biological screening of extracts from leaf and stem bark of Croton floribundus Spreng. (Euphorbiaceae)

Abstract This work describes the preliminary evaluation of cytotoxic, antimicrobial, molluscicidal, antioxidant and anticholinesterase activities from leaf (LECF) and stem bark alcoholic extracts (BECF) of the species Croton floribundus Spreng. (Euphorbiaceae), popularly known as capixingui or tapixingui. BECF presented significant toxicity (LC50 = 89.6 g/ml) in the Artemia salina Leach, 1819 (Crustacea: Branchiopoda) bioassay, whereas LECF did not show activity (LC50 > 1000 g/ml). From DPPH method, the values of IC50 for the LECF and BECF were 61.2 g/ml and 62.2 g/ml, respectively, showing that C. floribundus has an expressive antioxidant activity. Antimicrobial susceptibility was evaluated by microdilution technique and only BECF was active against Staphylococcus aureus (MIC = 39.6 g/ml). The extracts did not present molluscicidal activity against snail Biomphalaria glabrata Say, 1818 (Gastropoda: Planorbidae). Both extracts revealed the presence of several components with an inhibiting capacity of acetylcholinesterase enzyme on the bioautographic assay. C. floribundus showed to be a promising species considering that it exhibited good biological activity in the most assays performed.

Resumo Este trabalho descreve a avaliação preliminar das atividades citotóxica, antimicrobiana, moluscicida, antioxidante e anticolinesterásica de extratos alcoólicos das folhas (LECF) e das cascas do caule (BECF) da espécie Croton floribundus Spreng. (Euphorbiaceae), popularmente conhecida como capixingui ou tapixingui. No bioensaio com Artemia salina Leach, 1819 (Crustacea: Branchiopoda), BECF apresentou toxicidade significante (LC50 = 89,6 µg/ml), enquanto que LECF não apresentou atividade (LC50 > 1000 µg/ml). A partir do método de DPPH, os valores de IC50 para o LECF e BECF foram 61,2 µg/ml e 62,2 µg/ml, respectivamente, evidenciando que C. floribundus tem uma atividade antioxidante expressiva. A susceptibilidade antimicrobiana foi avaliada pela técnica de microdiluição e apenas BECF foi ativo contra Staphylococcus aureus (MIC = 39,6 mg/ml). Os extratos não apresentaram atividade moluscicida contra o caramujo Biomphalaria glabrata Say, 1818 (Gastropoda: Planorbidae). Ambos os extratos revelaram a presença de componentes com capacidade inibidora da enzima acetilcolinesterase no ensaio bioautográfico. C. floribundus mostrou ser uma espécie promissora considerando que exibiu boa atividade biológica na maioria dos ensaios testados.

Pneumatosis Cystoides Intestinales and Portomesenteric Venous Gas following Anticholinesterase Pesticide Poisoning

Pneumatosis cystoides intestinalis and portomesenteric venous gas are uncommon radiological findings, but are found commonly in cases of bowel ischemia, or as a result of various non-ischemic conditions. A 72-year-old man visited an emergency center with altered mental status 2 hours after ingestion of an unknown pesticide. On physical examination, he showed the characteristic hydrocarbon or garlic-like odor, miotic pupils with no response to light, rhinorrhea, shallow respiration, bronchorrhea, and sweating over his face, chest and abdomen. Laboratory results revealed decreased serum cholinesterase, as well as elevated amylase and lipase level. We made the clinical diagnosis of organophosphate poisoning in this patient based on the clinical features, duration of symptoms and signs, and level of serum cholinesterase. Activated charcoal, fluid, and antidotes were administered after gastric lavage. A computerized tomography scan of the abdomen with intravenous contrast showed acute pancreatitis, poor enhancement of the small bowel, pneumatosis cystoides intestinalis, portomesenteric venous gas and ascites. Emergent laparotomy could not be performed because of his poor physical condition and refusal of treatment by his family. The possible mechanisms were believed to be direct intestinal mucosal damage by pancreatic enzymes and secondary mucosal disruption due to bowel ischemia caused by shock and the use of inotropics. Physicians should be warned about the possibility of pneumatosis cystoides intestinalis and portomesenteric venous gas as a complication of pancreatitis following anticholinesterase poisoning.

Evaluation of Antioxidant, Anti-cholinesterase, and Anti-inflammatory Effects of Culinary Mushroom Pleurotus pulmonarius

Culinary mushroom Pleurotus pulmonarius has been popular in Asian countries. In this study, the anti-oxidant, cholinesterase, and inflammation inhibitory activities of methanol extract (ME) of fruiting bodies of P. pulmonarius were evaluted. The 1,1-diphenyl-2-picryl-hydrazy free radical scavenging activity of ME at 2.0 mg/mL was comparable to that of butylated hydroxytoluene, the standard reference. The ME exhibited significantly higher hydroxyl radical scavenging activity than butylated hydroxytoluene. ME showed slightly lower but moderate inhibitory activity against acetylcholinesterase (AChE) and butyrylcholinesterase than galantamine, a standard AChE inhibitor. It also exhibited protective effect against cytotoxicity to PC-12 cells induced by glutamate (10~100 µg/mL), inhibitory effect on nitric oxide (NO) production and inducible nitric oxide synthase protein expression in lipopolysaccharide-stimulated RAW 264.7 macrophages, and carrageenan-induced paw edema in a rat model. High-performance liquid chromatography analysis revealed the ME of P. pulmonarius contained at least 10 phenolic compounds and some of them were identified by the comparison with known standard phenolics. Taken together, our results demonstrate that fruiting bodies of P. pulmonarius possess antioxidant, anti-cholinesterase, and inflammation inhibitory activities.

Anticholinesterase and Anti-inflammatory Constituents from Beilschmiedia pulverulenta Kosterm

Phytochemical investigation from the stem bark of Beilschmiedia pulverulenta resulted in the isolation of five lignans, (+)-yangambin (1), (+)-sesartemin (2), (+)-excelsin (3), (+)-sesamin (4), and (+)-syringaresinol (5), together with lupeol (6), lupenone (7), β-sitosterol (8), and β-sitostenone (9). Their structures were established by the analysis of their spectroscopic (1D and 2D NMR) and spectrometric (MS) data, as well as by comparison with those reported in the literature. The isolated lignans were tested for their anticholinesterase (AChE: acetylcholine esterase and BChE: butyryl cholineesterase) and anti-inflammatory (COX-2: cyclooxygenase-2 and LOX: lipoxygenase) activities. All the isolated lignans (1 – 5) exhibited significant inhibition activities in AChE/BChE and COX-2/LOX assays with IC50 values ranging from 168.8 – 504.2 µM and 21.0 – 59.4 µM, respectively.

Antimicrobial (including antimollicutes), antioxidant and anticholinesterase activities of Brazilian and Spanish marine organisms – evaluation of extracts and pure compounds

Abstract This work describes the antimicrobial, antioxidant and anticholinesterase activities in vitro of organic extracts from fourteen seaweeds, eleven sponges, two ascidians, one bryozoan, and one sea anemone species collected along the Brazilian and Spanish coast, as well as the isolation of the diterpene (4R, 9S, 14S)-4α-acetoxy-9β,14α-dihydroxydolast-1(15),7-diene (1) and halogenated sesquiterpene elatol (2). The most promising antimicrobial results for cell wall bacteria were obtained by extracts from seaweeds Laurencia dendroidea and Sargassum vulgare var. nanun (MIC 250 μg/ml), and by the bryozoan Bugula neritina (MIC 62.5 μg/ml), both against Staphylococcus aureus. As for antimollicutes, extracts from seaweeds showed results better than the extracts from invertebrates. Almost all seaweeds assayed (92%) exhibited some antimicrobial activity against mollicutes strains (Mycoplasma hominis,Mycoplasma genitalium,Mycoplasma capricolum and Mycoplasma pneumoniae strain FH). From these seaweeds, A1 (Canistrocarpus cervicornis), A11 (Gracilaria sp.) and A4 (Lobophora variegata) showed the best results for M. pneumoniae strain FH (MIC 250 μg/ml). Furthermore, compounds 1 and 2 were also assayed against mollicutes strains M. hominis,M. genitalium,M. capricolum,M. pneumoniae strain 129 and M. pneumoniae strain FH, which showed MIC > 100 μg/ml. Antioxidant activities of extracts from these marine organisms were inactive, except for E7 (from sponge Ircinia sp.), which exhibited moderated antioxidant activities for two methods assayed (IC50 83.0 ± 0.1 μg/ml, and 52.0 ± 0.8 mg AA/g, respectively). Finally, for the anticholinesterase activity, all the 29 samples evaluated (100%) exhibited some level of activity, with IC50 < 1000 μg/ml. From these, seaweeds extracts were considered more promising than marine invertebrate extracts [A10 (IC50 14.4 ± 0.1 μg/ml), A16 (IC50 16.4 ± 0.4 μg/ml) and A8 (IC50 14.9 ± 0.5 μg/ml)]. The findings of this work are useful for further research aiming at isolation and characterization of active compounds.

Reduced expression of exocytotic proteins caused by anti-cholinesterase pesticides in Brachionus calyciflorus (Rotifera: Monogononta) / Redução da expressão de proteínas de exocitose causados por pesticidas anti-colinesterase em Brachionus calyciflorus (Rotifera: Monogononta)

AbstractThe organophosphate and carbamate pesticides methyl-parathion and carbaryl have a common action mechanism: they inhibit acetylcholinesterase enzyme by blocking the transmission of nerve impulses. However, they can alter the expression of exocytotic membrane proteins (SNARE), by modifying release of neurotransmitters and other substances. This study evaluated the adverse effects of the pesticides methyl-parathion and carbaryl on expression of SNARE proteins: Syntaxin-1, Syntaxin-4 and SNAP-23 in freshwater rotifer Brachionus calyciflorus. Protein expression of these three proteins was analyzed before and after exposure to these two pesticides by Western Blot. The expression of Syntaxin-1, Syntaxin-4 and SNAP-23 proteins in B. calyciflorussignificantly decreases with increasing concentration of either pesticides. This suggests that organophosphates and carbamates have adverse effects on expression of membrane proteins of exocytosis by altering the recognition, docking and fusion of presynaptic and vesicular membranes involved in exocytosis of neurotransmitters. Our results demonstrate that the neurotoxic effect of anticholinesterase pesticides influences the interaction of syntaxins and SNAP-25 and the proper assembly of the SNARE complex.

ResumoOs pesticidas organofosforados e carbamatos metil- paration e carbaril tem um mecanismo de ação comum: eles inibem a enzima acetilcolinesterase, bloqueando a transmissão dos impulsos nervosos. No entanto, eles podem alterar a expressão de proteínas de membrana de exocitose (SNARE), através da modificação da libertação de neurotransmissores e outras substâncias. Este estudo avaliou os efeitos adversos dos pesticidas metil- paration e carbaril sobre a expressão de proteínas SNARE: Sintaxina -1, Sintaxina-4 e SNAP-23 em rotíferos de água doce Brachionus calyciflorus. A expressão destas três proteínas foi analisada antes e depois da exposição a estes dois pesticidas por Western Blot. A expressão das proteínas Sintaxina-1, Sintaxina-4 e SNAP-23 em B. calyciflorus diminui significativamente com o aumento da concentração de ambos os pesticidas. Isto sugere que os organofosfatos e carbamatos têm efeitos adversos sobre a expressão de proteínas de membrana de exocitose, alterando o reconhecimento, de encaixe e fusão de membranas pré-sinápticas e vesiculares envolvidas na exocitose de neurotransmissores. Nossos resultados demonstram que o efeito neurotóxico de pesticidas anticolinesterásicos influencia a interação de sintaxinas e SNAP-25 e a montagem correta do complexo SNARE.

Tratamiento farmacológico de la enfermedad de Alzheimer y otras demencias / Pharmacological therapy of Alzheimer’s disease and other dementias

El aumento de la expectativa de vida trae a consecuencia el incremento de las enfermedades crónicas en el adulto mayor, prevaleciendo entre ellas, las demencias. El tratamiento de las mismas requiere de un enfoque multidimensional, pero en esta oportunidad nos centraremos en el tratamiento farmacológico. Una vez establecido el diagnóstico de demencia, es adecuado iniciar el tratamiento con fármacos anticolinesterásicos y/o memantina según corresponda. Si bien estos fármacos no revierten los síntomas cognitivos, enlentecen la progresión de la enfermedad. Debemos tener en cuenta en principio, las contraindicaciones, interacciones medicamentosas y monitorizar la aparición de efectos adversos. Durante el seguimiento del paciente se deberán aplicar tests funcionales y cognitivos para evaluar la eficacia de llos medicamentos prescriptos. En la actualidad, están en estudio fármacos pretendidamente curativos cuyo blanco es la sustancia amiloide y la proteína tau.

Increased life expectancy brings a result, the increase of chronic diseases in the elderly, prevailing including dementia. Treating them requires a multidimensional approach. This time we will focus on pharmacological treatment of them. Once the diagnosis of dementia, it is appropriate to initiate treatment with acetylcholinesterase inhibitors and/or memantine accordingly. While these drugs do not reverse the cognitive symptoms, slow the progression of the disease. We should note in principle, contraindications, drug interactions and monitor the occurrence of adverse effects. During follow-up the patient must apply functional and cognitive tests to assess the effectiveness of prescription medicines. Currently under study are supposedly curative medicine that targets amyloid and tau protein.

Effect of ethanolic extract of Coriandrum sativum L. on tacrine induced orofacial dyskinesia.

The effect of ethanolic extract of Coriandrum sativum L. seeds (100, 200 mg/kg) was studied on tacrine induced orofacial dyskinesia. Tacrine (2.5 mg/kg, i.p.) treated animals were observed for vacuous chewing movements (VCM), tongue protrusions (TP) and orofacial bursts (OB) for 1 h followed by observations for locomotor changes and cognitive dysfunction. Sub-chronic administration of Coriandrum sativum L. seed extract (E-CS) (100, 200 mg/kg, p.o., for 15 days significantly (P <0.05) decreased the tacrine induced VCM, TP and OB; and also significantly (P <0.05), increased locomotion and cognition compared to the tacrine treated group. Biochemical analysis revealed that tacrine administration significantly (P <0.05) decreased the levels of superoxide dismutase (SOD), Catalase (CAT), glutathione reductase (GSH) levels and also significantly (P <0.05) increased lipid peroxidation (LPO) as an index of oxidative stress, whereas sub-chronic administration of E-CS significantly (P <0.05) improved the antioxidant enzyme (i.e. SOD, CAT, and GSH) levels and also significantly (P <0.05) decreased lipid peroxidation (LPO). The results have demonstrated the protective role of ethanolic extract of Coriandrum sativum. L against tacrine induced orofacial dyskinesia.

In vitro biological screening of the anticholinesterase and antiproliferative activities of medicinal plants belonging to Annonaceae

The aim of this research was to investigate the antiproliferative and anticholinesterase activities of 11 extracts from 5 Annonaceae species in vitro. Antiproliferative activity was assessed using 10 human cancer cell lines. Thin-layer chromatography and a microplate assay were used to screen the extracts for acetylcholinesterase (AchE) inhibitors using Ellman's reagent. The chemical compositions of the active extracts were investigated using high performance liquid chromatography. Eleven extracts obtained from five Annonaceae plant species were active and were particularly effective against the UA251, NCI-470 lung, HT-29, NCI/ADR, and K-562 cell lines with growth inhibition (GI50) values of 0.04-0.06, 0.02-0.50, 0.01-0.12, 0.10-0.27, and 0.02-0.04 µg/mL, respectively. In addition, the Annona crassiflora and A. coriacea seed extracts were the most active among the tested extracts and the most effective against the tumor cell lines, with GI50 values below 8.90 µg/mL. The A. cacans extract displayed the lowest activity. Based on the microplate assay, the percent AchE inhibition of the extracts ranged from 12 to 52%, and the A. coriacea seed extract resulted in the greatest inhibition (52%). Caffeic acid, sinapic acid, and rutin were present at higher concentrations in the A. crassiflora seed samples. The A. coriacea seeds contained ferulic and sinapic acid. Overall, the results indicated that A. crassiflora and A. coriacea extracts have antiproliferative and anticholinesterase properties, which opens up new possibilities for alternative pharmacotherapy drugs.

In-vitro Screening for acetylcholinesterase enzyme inhibition potential and antioxidant activity of extracts of Ipomoea aquatica Forsk: therapeutic lead for Alzheimer’s disease.

Alzheimer’s disease (AD) is a primary degenerative disease of the central nervous system. The progression of Alzheimer’s disease will ultimately lead to dementia, behavioral and cognitive impairments. Increased level of the enzyme acetylcholinesterase AChE plays a key role in hydrolysis of the neurotransmitter Acetylcholine (ACh) which worsens the condition of cognitive dysfunction. Several drug of natural origin are known to possess AChE inhibition and antioxidant activity. The main objective of the present study is to evaluate AChE inhibition and antioxidant activity of the plant Ipomoea aquatica Forsk. Leaves of Ipomoea aquatica Forsk was extracted with Chloroform, n-Hexane, Ethanol and mixture of Ethanol: water (6:4) (hydro-alcoholic extract) using soxhlet extraction. All the four extracts were examined for In-vitro anti-cholinesterase by Ellman’s method and antioxidant activity by DPPH and Hydrogen peroxide radical scavenging assay. Results obtained from the study clearly demonstrates that all four extract has shown promising acetylcholinesterase inhibition activity in hydro alcoholic extract reveals the best inhibition potential with IC50 49.03 μg /ml. Similarly all the extracts projects significant antioxidant activity in DPPH assay with IC50 value ranging from 19.64 to 88.63 μg /ml and in Hydrogen peroxide assay with IC50 value ranging from 56.79 to 137.3 μg /ml.