Chronic Alcohol Intoxication and Cortical Ischemia: Study of Their Comorbidity and the Protective Effects of Minocycline

Chronic alcohol intoxication (CAI) increases both morbidity and mortality of stroke patients. Despite the high prevalence of CAI and ischemic stroke, studies addressing their comorbidity and/or protective alternatives remain scarce. Thus, the influence of CAI on both stroke outcome and minocycline treatment (recognized for its neuroprotective effect) was investigated. Female Wistar rats (35 days old) were treated with water or ethanol (6.5 g/kg/day, 22.5% w/v) for 55 days. Then, focal ischemia was induced by endothelin-1 in the motor cortex. Two hours later, four doses of 50 mg/kg of minocycline every 12 hours followed by five doses of 25 mg/kg every 24 hours were administered. Behavioral performance (open field and rotarod tests) and immunohistochemical (cellular density, neuronal death, and astrocytic activation) and biochemical (lipid peroxidation and nitrite levels) analyses were performed. CAI increased motor disruption, nitrite and lipid peroxidation levels, and neuronal loss caused by ischemia, whereas it reduced the astrogliosis. Minocycline was effective in preventing the motor and tissue damage caused by stroke. However, these effects were attenuated when CAI preceded stroke. Our data suggest that CAI beginning in adolescence contributes to a worse outcome in ischemic stroke survivors and reduces the benefits of minocycline, possibly requiring adjustments in therapy

Neurobehavioral and Antioxidant Effects of Ethanolic Extract of Yellow Propolis

Propolis is a resin produced by bees from raw material collected from plants, salivary secretions, and beeswax. New therapeutic properties for the Central Nervous System have emerged. We explored the neurobehavioral and antioxidant effects of an ethanolic extract of yellow propolis (EEYP) rich in triterpenoids, primarily lupeol and β-amyrin. Male Wistar rats, 3 months old, were intraperitoneally treated with Tween 5% (control), EEYP (1, 3, 10, and 30 mg/kg), or diazepam, fluoxetine, and caffeine (positive controls) 30 min before the assays. Animals were submitted to open field, elevated plus maze, forced swimming, and inhibitory avoidance tests. After behavioral tasks, blood samples were collected through intracardiac pathway, to evaluate the oxidative balance. The results obtained in the open field and in the elevated plus maze assay showed spontaneous locomotion preserved and anxiolytic-like activity. In the forced swimming test, EEYP demonstrated antidepressant-like activity. In the inhibitory avoidance test, EEYP showed mnemonic activity at 30 mg/kg. In the evaluation of oxidative biochemistry, the extract reduced the production of nitric oxide and malondialdehyde without changing level of total antioxidant, catalase, and superoxide dismutase, induced by behavioral stress. Our results highlight that EEYP emerges as a promising anxiolytic, antidepressant, mnemonic, and antioxidant natural product

Acidente vascular encefálico isquêmico na exposição crônica ao etanol: estudo pré-clínico da comorbidade e da resposta a minociclina

Stroke is the second largest cause of death in the world and the leading in Brazil, with 87% of strokes due to ischemic processes. Chronic ethanol consumption, usually beginning in adolescence, is recognized as an independent risk factor for increased morbidity and mortality by stroke. Although cases combining the two diseases are relatively common, there is no data in animals or clinical models demonstrating the quality or mechanisms of interaction between the two morbidities, nor its impact on therapeutic intervention. Considering the recent studies proposing minocycline as a new therapeutic tool for the treatment of stroke, this study aimed to investigate the interaction between the Chronic Alcoholic Intoxication (CAI) started in adolescence and the stroke in motor cortex of adult rats, and the effects of treatment with minocycline on this interaction, using behavioral, cellular and molecular parameters. Female Wistar rats (35 days-old) were chronically exposed to ethanol (6.5 g/kg/day, 22.5% w/v) or water for 55 days. One day after the end of the CAI focal ischemia was induced in motor cortex with the endothelin-1 (ET-1), followed by seven-day treatment with minocycline or saline. After this period, the animals were assayed with open field and rota rod tests. Immediately, animals were sacrificed and cortex was dissected for evaluation of nitrite and lipid peroxidation levels. In all groups, some animals were perfused and the motor cortex subjected to histological analysis to assess the damage, and immunohistochemical labeling to neuronal death (anti-NeuN), microglial/macrophage (anti-ED1) and astrocytes (anti-GFAP) activation. The ethanol intoxication from puberty to adulthood potentiated the damage caused by stroke, causing major losses in capacity to start and running movements as well as the strength and motor coordination compared to ischemic animals pretreated with water. These manifestations were accompanied by increased neuronal loss, reduced ED-1+ and GFAP+ cells and higher levels of nitrite and lipid peroxidation. Treatment with minocycline was effective in preventing/reverse motor deficits and tissue damage induced by focal ischemia, also inhibiting the increase in oxidative stress markers. The CAI either alone with succeeded by focal ischemia, harmed the outcome of treatment with minocycline. Our results indicate that heavy alcohol intoxication during adolescence exacerbates the motor deficit and tissue damage in animals subjected to focal ischemia. This process appears to be associated with microglia/astroglial activation, but mainly with oxidative stress. It also shows that the previous history of CAI started adolescence interferes significantly in the treatment of cerebral ischemia with minocycline.O acidente vascular encefálico (AVE) é a segunda maior causa de morte no mundo e a principal no Brasil, sendo que 87% dos AVE ocorrem por processos isquêmicos (AVEI). O consumo crônico de etanol, que se inicia geralmente na adolescência, é reconhecido como um fator de risco independente para a elevação da morbidade e mortalidade por AVEI. Apesar de que casos que combinem as duas patologias sejam relativamente frequentes, não há dados disponíveis em modelos animais ou clínicos que demonstrem a qualidade ou mecanismos de interação entre as duas morbidades, e tão pouco suas consequências sobre a intervenção terapêutica. Considerando então os recentes estudos que propõem a minociclina como nova ferramenta terapêutica para o tratamento do AVEI, este estudo teve por objetivo investigar a interação entre a Intoxicação Alcoólica Crônica (IAC) iniciada na adolescência e o AVEI em córtex motor na fase adulta em ratos, e os efeitos do tratamento com minociclina sobre esta interação, usando parâmetros comportamentais, celulares e moleculares. Ratas Wistar (de 35 dias de idade) foram expostas cronicamente a etanol (6,5 g/kg/dia, 22,5% m/v) ou água por 55 dias. Um dia após o fim da IAC, foi induzida isquemia focal no córtex motor com endotelina- 1 (ET-1), seguindo-se sete dias de tratamento com minociclina ou salina. Ao final deste período os animais foram testados em modelos de campo aberto e rota rod. A seguir, os animais foram sacrificados e o córtex dissecado para avaliação dos níveis de nitritos e de peroxidação lipídica. De cada grupo, alguns animais foram perfundidos e o córtex motor submetidos à analise histológica, para avaliação do dano, e histopatológica, para a morte neuronal (anti-NeuN), ativação microglial/macrófagica (Anti-ED1) e astrocitária (anti-GFAP). A intoxicação por etanol a partir da puberdade até a idade adulta potencializou os danos causados pela isquemia, causando grandes perdas na capacidade de iniciar e gerir os movimentos, bem como na coordenação e força motora em comparação aos animais isquêmicos pré-tratados com água. Estas manifestações foram acompanhadas de aumento da perda neuronal, redução do número de células ED1+ e GFAP+ e maiores níveis de nitritos e peroxidação lipídica. O tratamento com minociclina foi eficiente em prevenir/reverter as perdas motoras e danos teciduais induzidos pela isquemia focal, inibindo também a elevação dos marcadores de estresse oxidativo. A IAC tanto isoladamente como sucedida pela isquemia focal, modificaram o desfecho do tratamento com minociclina. Os nossos resultados indicam que a intoxicação com álcool durante a adolescência agrava o déficit motor e danos no tecido em animais sujeitos a isquemia focal no córtex motor. Este processo parece estar associado com a ativação microglial/ astrocitária, mas principalmente com o estresse oxidativo. Mostra ainda que o histórico prévio de IAC iniciado na adolescência interfere significativamente no tratamento da isquemia cerebral com minociclina

Aniba canelilla (Kunth) Mez (Lauraceae) Essential Oil: Effects on Oxidative Stress and Vascular Permeability

The present study aimed to investigate the antioxidant activity of Aniba canelilla (kunth) Mez (Lauraceae) essential oil (AcEO), exploring its potential for prevention and/or treatment of oxidative stress and associated inflammatory process. With this aim, Wistar rats (n = 6/group) were pre-treated intraperitoneally with saline (0.9%) or AcEO (2 or 5 mg/kg) for 5 days. One hour after the last dose, inflammation and oxidative stress were induced by carrageenan (0.3 mg/kg; ip.) administration. Total antioxidant capacity, reduced glutathione (GSH) and lipid peroxidation levels, protein concentration, and leukocyte migration were evaluated in peritoneal fluid. Lipid peroxidation was also evaluated in plasma. Carrageenan strongly reduced the peritoneal antioxidant capacity and GSH concentration, increasing peritoneal and plasma lipid peroxidation. It also promoted increased plasma leakage and leukocyte migration. Treatment with AcEO (2 and 5 mg/kg), whose major constituent was 1-nitro-2-phenylethane (77.5%), increased the peritoneal antioxidant capacity and GSH concentrations, and reduced lipid peroxidation, both peritoneal and plasma, thus inhibiting the carrageenan-induced oxidative imbalance. AcEO also reduced the carrageenan-induced plasma leakage and leukocyte migration. These data demonstrate the AcEO antioxidant activity and its ability to modulate plasma leakage and leukocyte migration, confirming its potential for treating diseases associated with inflammation and oxidative stress

Alveolar bone loss induced by chronic ethanol consumption from adolescence to adulthood in Wistar rats

93-97Though there are literature indicating the bone loss due to alcohol consumption, studies on the association between ethanol consumption and periodontal breakdown in animals are either scarce or have provided conflicting results. Here, we investigated the effects of chronic alcohol exposure from adolescence to adulthood on the alveolar bone in rats. Wistar rats were exposed to ethanol (6.5 g/kg/day) in a solution of 22.5% (w/v) or distilled water (control) by gavage from 35 days of age (adolescent) until 90 days (adulthood). Evaluation of the bone loss was performed using scanning electronic microscopy, in which the distances between the cement-enamel junction and the alveolar bone crest from the palatal side of the first molar mandibular were measured. The measurements obtained were tabulated and analyzed using Student’s t-test. Alcohol-treated group revealed greater bone loss in comparison to the control group. These findings indicate that heavy chronic alcohol exposure from adolescent to adulthood can induce alveolar bone loss in rats associated to absence of periodontitis. </span

Amazonian Plants: A Global Bibliometric Approach to <i>Petiveria alliacea</i> L. Pharmacological and Toxicological Properties

Petiveria alliacea L. (Phytolaccaceae) holds significant importance in the Amazon region, where it has been traditionally utilized in folk medicine. In this study, we conducted a comprehensive bibliometric analysis using conventional metrics, combined with a critical content review of its pharmacological and toxicological properties, to identify gaps in the existing literature that require further investigation. Our investigation identified a total of 55 articles that met the inclusion criteria for this study. Remarkably, Brazil emerged as the primary contributor within the scope of this review, indicating a strong presence of research from this country. Furthermore, professional scientific societies have played a pivotal role in facilitating the dissemination of scientific findings through specialist journals, fostering the sharing of research work within the community. Analysis of keyword co-occurrence revealed that “Petiveria alliacea”, “plant extract”, and “guatemala” were the most frequently encountered terms, indicating their significance within the literature. In terms of study designs, in vivo and in vitro were the predominant types observed, highlighting their prevalence in this field of study. Our study also identified a lack in knowledge yet to be investigated

Methylmercury plus Ethanol Exposure: How Much Does This Combination Affect Emotionality?

Mercury is a heavy metal found in organic and inorganic forms that represents an important toxicant with impact on human health. Mercury can be released in the environment by natural phenoms (i.e., volcanic eruptions), industrial products, waste, or anthropogenic actions (i.e., mining activity). Evidence has pointed to mercury exposure inducing neurological damages related to emotional disturbance, such as anxiety, depression, and insomnia. The mechanisms that underlie these emotional disorders remain poorly understood, although an important role of glutamatergic pathways, alterations in HPA axis, and disturbance in activity of monoamines have been suggested. Ethanol (EtOH) is a psychoactive substance consumed worldwide that induces emotional alterations that have been strongly investigated, and shares common pathophysiological mechanisms with mercury. Concomitant mercury and EtOH intoxication occur in several regions of the world, specially by communities that consume seafood and fish as the principal product of nutrition (i.e., Amazon region). Such affront appears to be more deleterious in critical periods of life, such as the prenatal and adolescence period. Thus, this review aimed to discuss the cellular and behavioral changes displayed by the mercury plus EtOH exposure during adolescence, focused on emotional disorders, to answer the question of whether mercury plus EtOH exposure intensifies depression, anxiety, and insomnia observed by the toxicants in isolation

Unravelling motor behaviour hallmarks in intoxicated adolescents: methylmercury subtoxic-dose exposure and binge ethanol intake paradigm in rats

Fundação Amazônia de Amparo a Estudos e Pesquisa do Pará (FAPESPA, grant number 160/2014 Edital 006/2014). Belém-Filho IJA and Nascimento AS received scholarships from Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Universidade Federal do Pará. Faculdade de Ciências Farmacêuticas. Laboratório de Farmacologia da Inflamação e do Comportamento. Belém, PA, Brazil.Universidade Federal do Pará. Faculdade de Ciências Farmacêuticas. Laboratório de Farmacologia da Inflamação e do Comportamento. Belém, PA, Brazil.Universidade Federal do Pará. Faculdade de Ciências Farmacêuticas. Laboratório de Farmacologia da Inflamação e do Comportamento. Belém, PA, Brazil.Universidade Federal do Pará. Faculdade de Ciências Farmacêuticas. Laboratório de Farmacologia da Inflamação e do Comportamento. Belém, PA, Brazil.Universidade Federal do Pará. Faculdade de Ciências Farmacêuticas. Laboratório de Farmacologia da Inflamação e do Comportamento. Belém, PA, Brazil.Universidade Federal do Pará. Faculdade de Ciências Farmacêuticas. Laboratório de Farmacologia da Inflamação e do Comportamento. Belém, PA, Brazil.Universidade Federal do Pará. Faculdade de Ciências Farmacêuticas. Laboratório de Farmacologia da Inflamação e do Comportamento. Belém, PA, Brazil.Universidade Federal do Pará. Instituto de Ciências Biológicas. Laboratório de Farmacologia Molecular. Belém, PA, Brazil.Universidade Federal do Pará. Faculdade de Ciências Farmacêuticas. Laboratório de Microbiologia e Imunologia Clinica. Belém, PA, Brazil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Toxicologia. Ananindeua, PA, Brasil.Universidade Federal do Pará. Faculdade de Ciências Farmacêuticas. Laboratório de Farmacologia da Inflamação e do Comportamento. Belém, PA, Brazil.Methylmercury (MeHg) is a hazardous environmental pollutant, affecting Amazon basin communities by anthropogenic activities. The exact safe level of MeHg exposure is unclear, despite the efforts of health international societies to avoid mercury (Hg) poisoning. Central nervous system is severely impacted by Hg intoxication, reflecting on motor impairment. In addition, alcohol has been associated to an overall brain damage. According to lifestyle of Amazon riverside communities, alcohol intake occurs frequently. Thus, we investigated if continuous MeHg exposure at low doses during adolescence displays motor deficits (experiment 1). In the experiment 2, we examine if the co-intoxication (i.e. MeHg plus ethanol exposure) during adolescence intensify motor damage. In the experiment 1, Wistar adolescent rats (31 days old) received chronic exposure to low dose (CELD) of MeHg (40 μg/kg/day) for 35 days. For the experiment 2, five sessions of alcohol binge drinking paradigm (3ON-4OFF; 3.0 g/kg/day) were employed associated to MeHg intoxication. Motor behaviour was evaluated by the open field, pole test, beam walking and rotarod paradigms. CELDS of MeHg display motor function damage, related to hypoactivity, bradykinesia-like behaviour, coordination deficits and motor learning impairment. Co intoxication of MeHg plus ethanol reduced cerebellar Hg content, however also resulted in motor behavioural impairment, as well as additive effects on bradykinesia and fine motor evaluation

Margaritaria nobilis L.F. (Phyllanthaceae): Ethnopharmacology and Application of Computational Tools in the Annotation of Bioactive Molecules

Margaritaria nobilis is a shrubby species widely distributed in Brazil from the Amazon to the Atlantic Rainforest. Its bark and fruit are used in the Peruvian Amazon for disinfecting abscesses and as a tonic in pregnancy, respectively, and its leaves are used to treat cancer symptoms. From analyses via UHPLC-MS/MS, we sought to determine the chemical profile of the ethanolic extract of M. nobilis leaves by means of putative analyses supported by computational tools and spectral libraries. Thus, it was possible to annotate 44 compounds, of which 12 are phenolic acid derivatives, 16 are O-glycosylated flavonoids and 16 hydrolysable tannins. Among the flavonoids, although they are known, except for kaempferol, which has already been isolated from this species, the other flavonoids (10, 14, 15, 21, 24&ndash;26, 28&ndash;30, 33&ndash;35, 40 and 41) are being reported for the first time in the genus. Among the hydrolysable tannins, six ellagitannins present the HHDP group (6, 19, 22, 31, 38 and 43), one presents the DHHDP group (5), and four contain oxidatively modified congeners (12, 20, 37 and 39). Through the annotation of these compounds, we hope to contribute to the improved chemosystematics knowledge of the genus. Furthermore, supported by a metric review of the literature, we observed that many of the compounds reported here are congeners of authentically bioactive compounds. Thus, we believe that this work may help in understanding future pharmacological activities

The Role of the Adenosine System on Emotional and Cognitive Disturbances Induced by Ethanol Binge Drinking in the Immature Brain and the Beneficial Effects of Caffeine

Binge drinking intake is the most common pattern of ethanol consumption by adolescents, which elicits emotional disturbances, mainly anxiety and depressive symptoms, as well as cognitive alterations. Ethanol exposure may act on the adenosine neuromodulation system by increasing adenosine levels, consequently increasing the activation of adenosine receptors in the brain. The adenosine modulation system is involved in the control of mood and memory behavior. However, there is a gap in the knowledge about the exact mechanisms related to ethanol exposure&rsquo;s hazardous effects on the immature brain (i.e., during adolescence) and the role of the adenosine system thereupon. The present review attempts to provide a comprehensive picture of the role of the adenosinergic system on emotional and cognitive disturbances induced by ethanol during adolescence, exploring the potential benefits of caffeine administration in view of its action as a non-selective antagonist of adenosine receptors