Bone impairment in adolescent female rats chronically exposed to ethanol

258-262Ethanol consumption has increased among teenagers worldwide considerably,including females. Long-term ethanol consumption in women has been reported to cause bone metabolism imbalance. However, only few studies are available on the impact of long-term ethanol consumptionon bone morphology during adolescence. Here, we report the effects of chronic ethanol consumption on bone structure in adolescent female rats. Twenty female Wistar rats (35 days old) received, by gavage, distilled water (control) or ethanol (6.5 g/kg/day, 22.5% w/v) once daily for 55 days. After ethanol administration, animals were perfused, and the femora were collected. Morphometric evaluations were performed by electron microscopy scanning. Femora length, cortical bone thickness and medullar bone diameter was measured. The results demonstrated that ethanol exposure during adolescence reduced the length of femurs, with a decrease of the anterior thickness, posterior thickness, and mid-lateral diameter (P<0.05). Thus, long-term ethanol intake may lead to alterations on bone morphometry, reducing the thickness of compact bone and femur length in adolescent females

Neurotoxicology of alcohol: a bibliometric and science mapping analysis

Alcohol consumption is common in many societies and has increased considerably, resulting in many socioeconomic and public health problems. In this sense, studies have been carried out in order to understand the mechanisms involved in alcohol consumption and related harmful effects. This study aimed to identify and map the knowledge and to perform bibliometric analysis of the neurotoxicology of alcohol based on the 100 most cited articles. A search was carried out in the Web of Science Core Collection database and information was extracted regarding the journal, authors, keywords, year of publication, number of citations, country and continent of the corresponding author. For each selected manuscript, the study design, alcohol exposure model, dose, period of exposure, and effect on the central nervous system and research hotspots were mapped. The journal with the highest number of publications was Alcoholism: Clinical and Experimental Research (n = 11 papers), the author who contributed the most was Crews FT (n = 8 papers), the studies had a total of 288 keywords and 75% of the publications were from the United States of America. The experimental studies evaluated the effects of prenatal and postnatal exposure and were conducted in rats and mice using doses ranging from 2.5 to 14 g/kg/day, with administration by subcutaneous, intraperitoneal, intragastric, or inhalation route or with free access through drinking bottles. Among the studies mapped, the oldest one (1989) aimed to understand the systemic damage and mechanisms of action involved, while the most recent focused on understanding the receptors and mechanisms involved in addiction, as well as genetic factors. Our results show the panorama of the most widespread scientific production in the scientific community on the neurotoxicology of ethanol, a high prevalence was observed in studies that addressed fetal alcohol syndrome and/or the effects of ethanol on neurodevelopment

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

Caracterização dos efeitos comportamentais, teciduais e bioquímicos da administração intermitente e episódica de EtOH em ratas da adolescência à fase adulta

The consumption of ethanol (EtOH) is enhanced particularly in adolescent female pubic. The EtOH intake and intermittent episodic own consumption rate around 3 times per week. The toxic effects of this kind of consumption is especially dangerous over the continuous consumption of EtOH followed due to the high dietary intakes of abstinence, causing major changes in the central nervous system (CNS) maturing in a short time consumption. Considering the epidemiological relevance and the harmful effects of EtOH on the oxidative balance, hormone production and neurotrophin CNS maturing, the aim of this study was to investigate the behavioural, tissue and biochemical responses derived from intermittent and episodic consumption of EtOH in rats in phase from adolescence to adulthood. Wistar female adolescents (n = 80) received by gavage, distilled water or EtOH (3 g/kg/day) for 3 consecutive days per week. The animals were assessed seven and a half hours after the last administration day 1, 4 and 8 weeks of episodes of binge drinking (37, 58 and 86 DPN, respectively), besides, a period of 14 days of abstinence was added after BD 8 (100 DPN) to evaluate the ability to reverse the CNS damage generated on it. The battery of behavioural tests consisted of spontaneous locomotor activity, object recognition, elevated plus maze, test pole, walking beam and rotarod. The animals were sacrificed and blood samples collected for evaluation of corticosterone levels of malondialdehyde, catalase activity, the activity of superoxide dismutase and glutathione content. Therefore, the hippocampus was dissected to quantify the immunocontent BDNF. The administration of EtOH reached average peak blood concentration of 197.4 mg / dL and the period of 7.5 hours after the last administration EtOH in acute binge blood concentration was 0.7 mg / dL. Thus, the animals underwent behavioural tests post-consumer EtOH, not under the drug effect. Consumption of EtOH in binge did not affect weight gain of adolescent animals into adulthood, however, reduced the exploratory locomotor activity, impaired motor coordination, balance and motor learning associated with bradykinesia, as well as loss in the mnemonic process and increased anxiety-like behaviour. These losses were accompanied by hormonal elevation of corticosterone, reduced hippocampal BDNF levels and systemic imbalance in the oxidative balance. Thus, it was possible to identify that the damage found on the similar behaviour to anxiety, short-term memory, bradykinesia and spontaneous locomotor activity appeared from EtOH post-consumption for three consecutive days, however, they showed no recovery or worsening of damage after repeated episodes. In contrast, there was recovery of short-term memory in object recognition task associated with the return of normal levels of BDNF in adulthood. Moreover, it showed worsening in motor learning in young adult phase followed by gradual and partial recovery after prolonged period of drug withdrawal, yet the loss of motor coordination and balance remained in adulthood.CNPq - Conselho Nacional de Desenvolvimento Científico e TecnológicoO consumo do etanol (EtOH) tem aumentado principalmente no púbico feminino adolescente. A ingestão de EtOH na forma intermitente e episódica possui frequência de consumo em torno de 3 vezes por semana. Os efeitos tóxicos dessa forma de consumo são especialmente perigosos em detrimento ao consumo contínuo de EtOH devido às altas doses ingeridas seguidas de abstinência, acarretando maiores alterações no sistema nervoso central (SNC) em maturação em pouco tempo de consumo. Considerando a relevância epidemiológica e as consequências nocivas do EtOH sobre o balanço oxidativo, produção hormonal e de neurotrofina no SNC em maturação, o objetivo deste estudo foi investigar as respostas comportamentais, teciduais e bioquímicas derivadas do consumo intermitente e episódico de EtOH em ratas na fase de adolescência à idade adulta. Ratos Wistar fêmeas adolescentes (n=80) receberam, por gavagem, água destilada ou EtOH (3 g/kg/dia) durante 3 dias consecutivos por semana. Os animais foram avaliados sete horas e meia após o último dia de administração em 1, 4 e 8 semanas de episódios de binge drinking (37, 58 e 86 DPN, respectivamente), além disso, foi adicionado um período de 14 dias de abstinência após 8 BD (100 DPN) a fim de avaliar a habilidade do SNC em reverter danos gerados sobre ele. A bateria de testes comportamentais foi constituída de atividade locomotora espontânea, reconhecimento de objetos, labirinto em cruz elevado, pole test, beam walking e rotarod. Os animais foram sacrificados e as amostras de sangue coletadas para avaliação dos níveis de corticosterona, de malondialdeído, atividade da catalase, atividade da superóxido-dismutase e conteúdo de glutationa. Por conseguinte, o hipocampo foi dissecado para quantificação do imunoconteúdo de BDNF. A administração de EtOH alcançou concentração sanguínea média de 197,4 mg/dL e no período de 7,5 horas após a última administração de EtOH em binge agudo, a concentração sanguínea foi de 0,7 mg/dL. Dessa forma, os animais realizaram os ensaios comportamentais pós-consumo de EtOH, não sob efeito da droga. O consumo de EtOH em binge não alterou o ganho de peso dos animais da adolescência à vida adulta, no entanto, reduziu a atividade locomotora exploratória, prejuízo na coordenação motora, equilíbrio e aprendizado motor associado à bradicinesia, assim como, prejuízo no processo mnemônico e aumento do comportamento relacionado à ansiedade. Estes prejuízos foram acompanhados de elevação hormonal de corticosterona, redução dos níveis de BDNF hipocampal e desequilíbrio no balanço oxidativo sistêmico. Dessa forma, foi possível identificar que os prejuízos encontrados sobre o comportamento semelhante à ansiedade, memória de curta duração, bradicinesia e atividade locomotora espontânea apareceram desde o pós-consumo de EtOH por três dias consecutivos, no entanto, não apresentaram recuperação nem piora do dano após repetidos episódios. Em contrapartida, houve recuperação da memória de curta duração na tarefa de reconhecimento do objeto associado ao retorno dos níveis normais de BDNF na idade adulta. Além disso, demonstrou piora no aprendizado motor na fase adulto jovem seguido de recuperação gradual e parcial após período prolongado de retirada da droga, mesmo assim, o prejuízo da coordenação motora e equilíbrio permaneceram na fase adulta

Immunohistochemical changes and atrophy after chronic ethanol intoxication in rat salivary glands

Alcoholism in humans is a chronic and progressive disease, characterized by loss of ethanol consumption control. Previous studies have reported that prolonged exposure to ethanol was responsible for alterations in glandular tissues of human and rodents. However, the interrelationship between ethanol and the glandular system is still the subject of numerous investigations, including the possible resistance of the submandibular gland (SG). In the present study, we investigated whether chronic ethanol exposure during adolescence may affect the parotid gland (PG) and SG in female rats. Female rats (n=16) were treated with distilled water or ethanol (dose of 6.5 g/kg/day, 22.5% w/v) through gavage for 55 days. Glands were collected, weighed and submitted to histological processing. Morphometric analysis was assessed by parenchymal and stromal area measurements. Smooth muscle actin (α-SMA), cytokeratin-19 (CK19) and apoptotic caspase3 (CAS) were measured using ImageJ® software. Chronic ethanol administration did not alter the body weight of rats after treatment, although it increased glandular weight (p<0.001), reduced the parenchyma area (p<0.001) and decreased CK19 and α-SMA immunostainning in the PG. Besides, ethanol induced CK19 and CAS overexpression, and the occurrence of duct-like structures in SG. These results suggest that ethanol induces histological and morphometric changes in salivary glands of female rats intoxicated with ethanol during adolescence. Furthermore, the mechanism underlying these alterations needs to be investigated but may be not related to the inflammatory process

Morfologia radicular da dentição permanente de Sapajus apella: morfometria, anatomia macroscópica, ultraestrutura e propriedades físicas

O objetivo desse trabalho foi contribuir com o estudo anatômico, morfométrico, ultraestrutural e propriedades físicas dos dentes permanentes do primata Sapajus apella. Para tal, foram utilizados 10 animais adultos e machos. Os dentes foram avaliados quanto ao seu comprimento e quanto à anatomia radicular externa e interna considerando número de raízes e canais, forma e direção radicular e forma do canal, assim como análise da densidade e diâmetro tubular do canal radicular, composição e microdureza dentinária. A anatomia radicular desse primata apresentou especificidades, como o número de raízes do segundo pré-molar superior e a presença do terceiro pré-molar. Quanto à densidade e o diâmetro dos túbulos dentinários, observou-se uma diminuição do número e diâmetro de túbulos ao longo do canal radicular, havendo diferença estatisticamente significante ao se comparar o terço apical com as regiões cervical e média, padrão de densidade e tamanho semelhantes a dentes humanos. Semelhanças também foram encontradas com dentes humanos quando comparados os valores de microdureza e proporção de elementos químicos encontrados na dentina radicular

Contributing to Understand the Crosstalk between Brain and Periphery in Methylmercury Intoxication: Neurotoxicity and Extracellular Vesicles

Human exposure to methylmercury (MeHg) is currently high in regions such as the Amazon. Understanding the molecular changes associated with MeHg-induced neurotoxicity and the crosstalk with the periphery is essential to support early diagnoses. This work aimed to evaluate cellular and molecular changes associated with behavioral alterations in MeHg acute exposure and the possible changes in extracellular vesicles (EVs) number and S100β content. Adults male Wistar rats were orally treated with 5 mg/kg for four days. Behavioral performance, molecular and histological changes in the cerebellum, and plasma EVs were assessed. MeHg-intoxicated animals performed significantly worse in behavioral tests. MeHg increased the number of GFAP+ cells and GFAP and S100β mRNA expression in the cerebellum but no change in NeuN+ or IBA-1+ cells number was detected. The number of exosomes isolated from plasma were decreased by the metal. S100B mRNA was detected in circulating plasma EVs cargo in MeHg exposure. Though preliminary, our results suggest astrocytic reactivity is displaying a protective role once there was no neuronal death. Interestingly, the reduction in exosomes number could be a new mechanism associated with MeHg-induced neurotoxicity and plasma EVs could represent a source of future biomarkers in MeHg intoxication

Binge Drinking of Ethanol during Adolescence Induces Oxidative Damage and Morphological Changes in Salivary Glands of Female Rats

This study investigates morphological and biochemistry effects of binge ethanol consumption in parotid (PG) and submandibular (SG) salivary glands of rats from adolescence to adulthood. Female Wistar rats (n=26) received ethanol at 3 g/kg/day (20% w/v) for 3 consecutive days/week from the 35th until the 62nd day of life. Animals were treated in two periods: 1 week (G1) and 4 weeks (G2), with a control (treated with distilled water) and an ethanol group to each period. In morphological analysis, morphometric and immunohistochemistry evaluation for smooth muscle actin (αSMA), cytokeratin-18 (CK-18), and vimentin (VIM) were made. Biochemical changes were analyzed by concentration of nitrites and levels of malondialdehyde (MDA). The difference between groups in each analysis was evaluated by Mann-Whitney U test or Student’s t-test (p≤0.05). PG showed, at one week of ethanol exposure, lower CK-18 and α-SMA expression, as well as MDA levels. After four weeks, lower CK-18 and higher MDA levels were observed in PG exposed to ethanol, in comparison to control group. SG showed lower α-SMA expression after 1 and 4 weeks of ethanol exposure as well as higher MDA levels after 1 week. Ethanol binge consumption during adolescence promotes tissue and biochemical changes with only one-week binge in acinar and myoepithelial PG cells