Adiponectin secretion by perivascular adipose tissue supports impaired vasodilation in a mouse model of accelerated vascular smooth muscle cell and adipose tissue aging.

OBJECTIVE: Perivascular adipose tissue (PVAT) function during aging has not been investigated in detail so far and its effect on vasodilation remains to be fully elucidated. The aim of this study was to investigate endothelium-dependent vasodilation of thoracic aorta in a mouse model of accelerated, selective vascular smooth muscle and PVAT aging, induced by SM22α-Cre-driven genetic deletion of the endonuclease ERCC1 (SMC-KO mice) versus healthy littermates (LM). We hypothesized that PVAT enhances vasodilation in LM, possibly through adiponectin secretion, which might be compromised in SMC-KO animals. METHODS: Thoracic aorta was isolated from SMC-KO animals and LM and segments with and without PVAT were mounted in wire myography setups. The endothelium-dependent vasodilation was assessed via acetylcholine dose-response curves and pathway contribution was studied. Moreover, adiponectin secretion was measured after stimulating the aortic segments with PVAT with acetylcholine. RESULTS: Adiponectin, secreted by PVAT, led to increased NO-contribution to endothelium-dependent vasodilation in healthy LM, although this did not increase maximum relaxation due to loss of EDH. Endothelium-dependent vasodilation was decreased in SMC-KO animals due to reduced NO-contribution and complete EDH loss. Despite strong lipodystrophy the PVAT partially compensated for lost vasodilation in SMC-KO. LM PVAT contained acetylcholinesterase that attenuated acetylcholine responses. This was lost in SMC-KO. CONCLUSIONS: PVAT-derived adiponectin is able to partially compensate for age-related decline in NO-mediated vasodilation, even during strong lipodystrophy, in conditions of absence of compensating EDH. In aorta with healthy PVAT acetylcholinesterase modulates vascular tone, but this is lost during aging, further compensating for decreased acetylcholine responsiveness. Thus, preservation of adiponectin levels, through relatively increased production in lipodystrophic PVAT, and reduction of cholinesterase might be regulatory mechanisms of the PVAT to preserve cholinergic vasodilation during aging.

Neuroprotective effects of cordycepin inhibit glutamate-induced apoptosis in hippocampal neurons.

Glutamate is a neurotransmitter that can cause excitatory neurotoxicity when its extracellular concentration is too high, leading to disrupted calcium balance and increased production of reactive oxygen species (ROS). Cordycepin, a nucleoside adenosine derivative, has been shown to protect against excitatory neurotoxicity induced by glutamate. To investigate its potential neuroprotective effects, the present study employed fluorescence detection and spectrophotometry techniques to analyze primary hippocampal-cultured neurons. The results showed that glutamate toxicity reduced hippocampal neuron viability, increased ROS production, and increased intracellular calcium levels. Additionally, glutamate-induced cytotoxicity activated acetylcholinesterase and decreased glutathione levels. However, cordycepin inhibited glutamate-induced cell death, improved cell viability, reduced ROS production, and lowered Ca2+ levels. It also inhibited acetylcholinesterase activation and increased glutathione levels. This study suggests that cordycepin can protect against glutamate-induced neuronal injury in cell models, and this effect was inhibited by adenosine A1 receptor blockers, indicating that its neuroprotective effect is achieved through activation of the adenosine A1 receptor.

Entomopathogenic fungi based microbial insecticides and their physiological and biochemical effects on Spodoptera frugiperda (J.E. Smith).

Background: 'The fall armyworm, Spodoptera frugiperda', represents a significant threat to maize production, a major staple crop in Asian countries. Methods: In pursuit of more effective control of this insect pest, our study assessed the physiological and biochemical effects of the entomopathogenic fungus Metarhizium anisopliae against the larvae of S. frugiperda. Results: Results revealed that, following nine days of treatment, a high concentration of conidia (1.5x107 conidia/mL-1) was toxic to all stages of larvae (second to fifth instar), resulting in 97% mortality of the second instar, 89% mortality of the third instar, 77% mortality of the fourth instar, and 72% mortality of fifth instar. All larval instars were found to have dose-dependent mortality effects. Treated S. frugiperda larvae further displayed significant physiological, morphological, and behavioral changes. Here, treated larvae displayed significantly lower levels of acetylcholinesterase, α-carboxylesterase, and ß-carboxylesterase enzyme activity when compared to control groups. Treated larvae underwent an outward morphological change as the result of a decrease in the exterior cuticle of the anal papillae and a demelanization of the interior cuticle. Treated larvae also exhibited abnormal feeding behaviors as a consequence of the negative impact of conidia treatment on the neuromuscular system. Investigation into the effect of M. anisopliae on the non-target organism, the earthworm Eudrilus eugeniae, revealed that M. anisopliae conidia did not produce significant pathogenicity following three days of treatment. Furthermore, histological analysis revealed no significant effect of the entomopathogenic fungi on the gut tissue of the non-target organism. Conclusion: This study highlights the potential of M. anisopliae in the control of S. frugiperda.

Cajanus cajan (L) Millsp. seed extract ameliorates scopolamine-induced amnesia through increase in antioxidant defense mechanisms and cholinergic neurotransmission.

Decline in cholinergic function and oxidative/nitrosative stress play a central role in Alzheimer's disease (AD). Previous quantitative HPLC profiling analysis has revealed the presence of Pinostrobin, formononetin, vitexin and other neuroprotective flavonoids in Cajanus cajan seed extract. This study was designed to investigate the protective action of Cajanus cajan ethanol seed extract (CC) on learning and memory functions using scopolamine mouse model of amnesia. Materials and methods: Adult mice were pretreated with CC (50, 100, or 200mg/kg, p.o) or vehicle (10ml/kg, p.o) for 16 days consecutively. Scopolamine, a competitive muscarinic cholinergic receptor antagonist (1mg/kg, i.p.) was given an hour after CC pretreatment from days 3 to 16.  The mice were subjected to behavioural tests from day 11 (open field test (OFT)/ Y-maze test (YMT) and Morris water maze task (MWM) from days 12-16. Animals were euthanized 1h after behavioral test on day 16 and discrete brain regions isolated for markers of oxidative stress and cholinergic signaling. Molecular docking analysis was undertaken to predict the possible mechanism(s) of CC-induced anti-amnesic action.  pre-administration of CC significantly reversed working memory and learning deficits caused by scopolamine in YMT and MWM tests, respectively. Moreover, CC prevented scopolamine-induced oxidative and nitrosative stress radicals in the hippocampus evidenced in significant increase in glutathione (GSH) level, superoxide dismutase (SOD) and catalase (CAT) activities with a marked decrease in malondialdehyde (MDA) production, as well as significant inhibition of hippocampal scopolamine-induced increase in acetylcholinesterase activity by CC. The molecular docking analysis showed that out of the 19 compounds, the following had the highest binding affinity; Pinostrobin (-8.7 Kcal/mol), friedeline (-7.5kCal/mol), and lupeol (-8.2 Kcal/mol), respectively, to neuronal muscarinic M1 acetylcholine receptor, α7 nicotinic acetylcholine receptor and amyloid beta peptide binding pockets, which further supports the ability of CC to enhance neuronal cholinergic signaling and possible inhibition of amyloid beta aggregation. This study showed that Cajanus cajan seeds extract improved working memory and learning through enhancement of cholinergic signaling, antioxidant capacity and reduction in amyloidogenesis.

Effect of standardized Eucalyptus globulus leaf extract on brain oxidative stress and aberrant neurochemistry of fructose-streptozotocin-induced diabetic rats.

The neuro-pharmacological effect of Eucalyptus globulus ethanol leaf extract in fructose-streptozotocin-induced diabetic rats was evaluated in this study. The phytochemical analysis of the extract was carried out using HPLC-DAD. Diabetes was induced in rats with 10% fructose in drinking water and a single intraperitoneal injection of 40 mg/kg streptozotocin (STZ). Diabetic animals were orally treated with 100-400 mg/kg of the extract for 21 days with glibenclamide as the reference drug. Blood and brain tissue were processed for the determination of serum electrolyte levels, hematological indices, and biochemical estimations. Ergosterol, pinitol, catechin, quercetin, robinetinidol, and other polyphenols were identified in the extract. Diabetic animals showed decreased serum potassium and sodium ion levels and decreased hematocrit, hemoglobin, red blood cells, white blood cells and lymphocytes but increased neutrophils. The brains of animals in the untreated diabetic group with increased blood glucose level showed oxidative stress (increased level of MDA and myeloperoxidase but decreased level of reduced glutathione and superoxide dismutase) and disturbed neurochemistry (increased level of acetylcholinesterase and monoamine oxidase but decreased level of Na+K+ATPase, tyrosine hydroxylase and dopamine). Administration of the Eucalyptus globulus leaf extract remarkably ameliorated the observed hyperglycemia, electrolyte, and hematological imbalances in animals. In addition, the administration of the extract attenuated the brain redox imbalance, and neurochemical disturbances in the rats. These results show that Eucalyptus globulus leaves contain antioxidant and neurotransmitter modulating phytochemicals with the potential to be developed as therapeutic agents for the management of diabetic cerebrovascular problems and related complications.

Anticholinesterase activity of ß-carboline-1, 3, 5-triazine hybrids

Abstract The ß-carboline-1,3,5-triazine hydrochlorides 8-13 were evaluated in vitro against acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE). The analysed compounds were selective to BuChE, with IC50 values in the range from 1.0-18.8 µM being obtained. The N-{2-[(4,6-dihydrazinyl-1,3,5-triazin-2-yl)amino]ethyl}-1-phenyl-ß-carboline-3-carboxamide (12) was the most potent compound and kinetic studies indicate that it acts as a competitive inhibitor of BuChE. Molecular docking studies show that 12 strongly interacts with the residues of His438 (residue of the catalytic triad) and Trp82 (residue of catalytic anionic site), confirming that this compound competes with the same binding site of the butyrylthiocholine

Exposição ambiental/ocupacional aos agrotóxicos em gestantes residentes em um município rural / Environmental/occupational exposure to pesticides of pregnant women living in a countryside municipality / Exposición ambiental/ocupacional a los agrotóxicos en gestantes residentes en un municipio rural

Objective: The study's goal has been to analyze if environmental or occupational exposure to pesticides can produce changes in pregnant women living in a countryside municipality. Methods: The participants of this study were twenty-three pregnant women, who both answered a questionnaire and donated biological material in order to perform Micronucleus (MN) Tests in lymphocytes, oral epithelial cells, and also for measuring the enzyme activity of erythrocyte acetylcholinesterase. Results: Considering the total analyzed samples, the following was found: an average of 8 ± 2.92 MN/1000 oral epithelial cells from urban participants; an average of 6.82 ± 3.43 MN/1000 oral epithelial cells from rural participants; and 100% of the microscope slides contained cells with two MN, which shows high intensity lesions to the DNA. There was found a high frequency of spontaneous abortions (34.8%), greater than in Brazil. Conclusion: The exposure of pregnant women living in a countryside municipality to pesticides may increase the rate of spontaneous abortions, as well as the chances of mutagenic effects

Objetivo: Analisar se a exposição ambiental ou ocupacional aos agrotóxicos causa alterações em gestantes residentes em um município rural. Métodos: Compuseram a amostra 23 gestantes, que responderam a um questionário e doaram amostras biológicas para a realização dos testes de micronúcleos (MN) em linfócitos, em células do epitélio oral, e para a dosagem da atividade da enzima acetilcolinesterase eritrocitária. Resultados: Obteve-se uma média de 8 ± 2,92 MN/1000 células do epitélio oral analisadas em amostras de participantes da zona urbana, 6,82 ± 3,43 MN/1000 de participantes da zona rural, e 100% das lâminas continham células com dois MN, o que demonstra lesões ao DNA de maior intensidade. Encontrou-se uma frequência elevada de casos de abortos espontâneos (34,8%), superior à encontrada no Brasil. Conclusão: A exposição de gestantes residentes em um município rural aos agrotóxicos eleva a taxa de abortos espontâneos, bem como as chances de ocorrência de efeitos mutagênicos

Objetivo: Analizar si la exposición ambiental o ocupacional a los agrotóxicos causa cambios en gestantes residentes en un municipio rural. Métodos: Compusieron la muestra 23 gestantes, que respondieron a un cuestionario y donaron muestras biológicas para la realización de las pruebas de micronúcleos (MN) en linfocitos, en células del epitelio oral, y para la dosificación de la actividad de la enzima acetilcolinesterasa eritrocitaria. Resultados: Se obtuvieron una media de 8 ± 2,92 MN / 1000 células del epitelio oral analizadas en muestras de participantes de la zona urbana, 6,82 ± 3,43 MN / 1000 de participantes de la zona rural, y el 100% de las láminas contenían células con dos MN, lo que demuestra lesiones al ADN de mayor intensidad. Se encontró una frecuencia elevada de casos de abortos espontáneos (34,8%), superior a la encontrada en Brasil. Conclusión: La exposición de gestantes residentes en un municipio rural a los agrotóxicos eleva la tasa de abortos espontáneos, así como las posibilidades de ocurrencia de efectos mutagênicos

Avaliação "in vitro" do efeito da infusão de Cunila microcephala Benth sobre a atividade da enzima acetilcolinesterase e biomarcadores de estresse oxidativo em eritrócitos de agricultores / Effect of the Cunila microcephala Benth infusion on the activity of Acetilcolinesterase enzyme and on the biomarkers of oxidative stress in farmers erythrocytes

RESUMO Tivemos como objetivo avaliar o efeito da infusão de Cunila microcephala Benth sobre a atividade da enzima acetilcolinesterase (AChE) e marcadores de estresse oxidativo em eritrócitos de agricultores. Foram utilizadas amostras provenientes de 16 trabalhadores rurais expostos a pesticidas agrícolas pelo período mínimo de 5 anos e um grupo controle constituído de 16 indivíduos não expostos a agrotóxicos. As hemácias dos agricultores e o grupo A foram expostos “in vitro” à solução salina (NaCl 0,9%). Os demais grupos foram expostos à infusão de poejo nas concentrações de 0; 5; 10; 25 e 50 g/L (Grupos B; C; D e E, respectivamente). Em seguida, foram realizadas as determinações da atividade da AChE e dos níveis de substâncias reativas ao ácido tiobarbitúrico (TBARS), proteínas carboniladas (PCs) e glutationa reduzida (GSH). Os resultados mostram que a infusão de poejo 50g/L, aumenta a atividade da enzima AChE e os níveis de GSH. Contudo, os níveis de TBARS e PCs diminuíram após o tratamento com a infusão de poejo 25 e 50 g/L. A infusão de poejo, na concentração de 50 g/L, é capaz de reverter, “in vitro” a inibição da atividade da AChE que ocorre pela exposição a pesticidas, e ainda demonstra um importante potencial antioxidante, tendo em vista que diminuiu danos lipídicos e proteicos e ainda, estimulou a produção do principal antioxidante não enzimático endógeno.

ABSTRACT Evaluating the effect of infusion Cunila microcephala Benth on acetylcholinesterase activity (AChE) enzyme and on biomarkers of oxidative stress in farmers erythrocytes. We used samples from 16 rural workers exposed to pesticides for a minimum of five years, and a control group composed of 16 individuals not exposed to pesticides. The erythrocytes of farmers and from group A were exposed “in vitro” the saline solution (NaCl 0,9%). The other groups were exposed to the infusion of “poejo” at concentrations of 0; 5; 10; 25 and 50 g/L (Groups B, C, D and E, respectively). Then, it was realized the analitical determinations of AChE activity and TBARS, PCs and GSH levels. The results showed that “poejo” infusion 50g/L, increased the AChE activity and GSH levels. However, the TBARS e PCs levels decreased after the treatment with “poejo” infusion 25 e 50 g/L. The “poejo” infusion 50 g/L is able to revert “in vitro” the inhibition of AChE activity that occurs by exposure to pesticides and also demonstrates an important antioxidant potential, given that decreased lipid and protein damage and also it stimulated the production of the main non-enzymatic antioxidant endogenous.

Correlação entre as atividades antiradical, antiacetilcolinesterase e teor de fenóis totais de extratos de plantas medicinais de farmácias vivas / Correlation between the anti-radical, anti-acetylcholinesterase activities and phenolic content of some extracts of live herbal pharmacies in the Northeast of Brazil

O objetivo deste trabalho foi avaliar a correlação entre o conteúdo de fenóis de extratos de 18 plantas medicinais comuns nas Farmácias Vivas com a atividade sequestradora do radical livre DPPH e com a inibição da enzima acetilcolinesterase. Maiores conteúdos de fenóis totais foram encontrados nas plantas: Eugenia uniflora, Lippia alba, Lippia microphylla, Mentha x villosa, Ocimum gratissimum e Ocimum selloi. As espécies que apresentaram maior atividade antioxidante, semelhante a quercetina, foram: Spondias mombim, Malphigia glabra, L. alba e Croton zenhtneri,. As plantas Cecropia pachystachia, L. alba, L. microphylla, M. glabra, O. gratissimum e Plectranthus ornatus mostraram maiores halos de inibição da enzima acetilcolinesterase. Combinando as duas ações, as plantas mais ativas foram a M. glabra e a L. alba e somente a segunda correlaciona-se diretamente com o maior teor de fenóis totais. Através deste estudo, M. glabra e L. alba foram consideradas as plantas mais promissoras para estudos subsequentes a fim de encontrar novos compostos com ação potencial contra a doença de Alzheimer.

The purpose of this study was to evaluate the phenolic content of 18 medicinal plant extracts of "Live Pharmacies" and correlate them with their free radical scavenging activity and inhibition of the acetylcholinesterase enzyme. Higher amounts of phenolic compounds were found in the following plants: Eugenia uniflora, Lippia alba, Lippia microphylla, Mentha x villosa, Ocimum gatissimum, and Ocimum selloi. The species that showed higher anti-radical activity were Croton zenhtneri, Lippia alba, Malphigia glabra, and Spondias mombim, similar to quercetin. The plants Cecropia pachystachia, L. alba, L. microplylla, M. glabra, O. gratissimum, and Plectranthus ornatus showed larger inhibition zones of the acetylcholinesterase enzyme. By correlating the two actions L. alba and M. glabra are common as more active to both tests and only L. alba correlates directly with the phenolic content. Through this study, L. alba and M. glabra are considered the most promising plants for further studies to find new compounds with potential action against Alzheimer's disease.

Enfermedad de Alzheimer: Tratamientos actuales y futuros / Alzheimer's Disease: Current and Future Treatments

Unas 25 millones de personas padecen enfermedad de Alzheimer en el mundo, y probablemente en los práximos 20 años, se registrarán unos 70 millones de nuevos casos. Caracterizaada por una pérdida progresiva de la memoria, el déficit de la capacidad cognitiva es proporcional a la densidad de placas seniles, a la acumulación de la proteína beta amiloide, degeneraciones neuríticas y ovillos neurofibrilares, particularmente en el hipocampo y en la corteza cerebral. Este cuadro histopatológico se asocia a otro neuroquímico, caracterizado por una disminución de las enzimas colina acetiltransferasa y acetilcolinesterasa, y una menor densidad de los receptores colinérgicos muscarínicos y nicotínicos. Ello ha generado la teoría colinérgica del Alzheimer, que ha dado lugar a una aproximación racional al tratamiento de la enfermedad. Hoy disponemos de 3 anticolinesterásicos, galantamina, donepecilo y rivastigmina aprobados por FDA, que se recomiendan en EA leves y moderadas y de un antagonista no competitovo de los receptores NMDA memantina EA. El beneficio es modesto en relación a lo cogitivo y conductual. Se incluyen estudios sobre recomendaciones, farmacología, farmacognética, eficacia, tolerancia, características de los pacientes respondedores a los diferentes anticolinesterásicos, sus reemplazos el uso de comprimidos, parches, beneficiios, efectos adversos y de las nuevas terapéuticas que están en desarrollo como estanercept, NP12, resveratrol PBT2 y vacuna nasal, entre otras...

About 25 millions of individuals in the world suffer from Alzheimer's disease. The next 20 years shall probably register 70 millions of new cases. Characterized by a progressive loss of memory, cognitive deficit is proportional to the density of senile plaques, accumulation of beta amyloid, neuritic degeneration and neurofibrillary tangles, particularly in the hippocampus and cerebral cortex. This histopathological condition is associated with another neurochemical, chracterized by a decrease in choline acetyltransferase and acetylcholinesterase enzymes, and a lower density of muscrinic and nicotinic cholinergic receptors. This results in the cholinergic theory of Alzheimer's, which has led to a rational approach to treatment of disease. Today we have 3 anticholinesterase Galantamine, Donepzil and Rivastigmine approved by FDA recommended in mild to moderate AD and an uncompetitive antagonist of NMDA receptors Memantine is recommended in mild Alzheimer's disease for people who can not take ACE inhibitors, and severe AD The benefit is modest in realtion to the cognitive and behavioral. Studies are included on therapeutic recommendations, pharmacology, pharmacogenetic, efficiency, tolerance, characteristics of responders to different anticholinesterases, their relacements, the use of pills, patches, benefits, side effects and new therapeutic under development as Etanercept, NP12, PBT 2, Resveratrol, Nasal Vaccine, among others...