Teste de marcha fisiológica: um método eficaz para analisar distúrbios de equilíbrio, locomoção e neuromusculares em roedores

A avaliação da atividade locomotora animal é uma ferramenta comportamental bastante utilizada para mensurar os mecanismos subjacentes a uma determinada doença, distúrbio ou lesão e efeitos da exposição a um xenobiótico. Um dos testes mais utilizados em roedores para avaliar o equilíbrio e coordenação motora é o teste da trave elevada que, apesar de ser um teste barato e que exige um aparato simples, é necessário um longo período de treino e habituação dos animais. O desenvolvimento e caracterização de um teste alternativo, chamado de teste da marcha, tem o potencial de contornar o tempo e o esforço necessários ao treino dos animais, considerando-o um método eficaz, barato e rápido para a análise de comportamentos avaliados comparativamente pelo alto teste de feixe. Portanto, o presente estudo concentrou-se em determinar a eficácia e viabilidade do teste de marcha para avaliação da locomoção e equilíbrio de roedores em substituição ao teste da trave elevada. Para isso, ratos machos foram divididos em 3 grupos, sendo 1 grupo controle exposto à solução salina (NaCl 0,9%) e 2 grupos experimentais expostos à dose única de 0,2 e 1,0 mg/kg de ivermectina por via intraperitoneal para indução da alteração locomotora. Os testes de trave elevada e marcha foram realizados 15 min e 24h após a administração da droga. Os resultados mostram que os grupos experimentais tiveram dificuldade em realizar as tarefas de qualquer teste em ambos os momentos analisados em comparação com os grupos de controle. Na trave elevada, os animais experimentais tiveram dificuldade em manter o equilíbrio e andar. No teste de marcha, os animais experimentais apresentaram alterações na marcha, que foram quantificadas por: (a) encurtamento do comprimento da passada, (b) diminuição da passada, (c) alteração da simetria da passada e (d) alteração da área da passada. Tais resultados são indicativos de esforços compensatórios e foram comparáveis entre os dois testes. Em conjunto, os dados indicam que o teste de marcha atende a todos os requisitos para avaliação da coordenação motora em roedores. O teste de marcha é, portanto, validado como um complementar para o teste da trave elevada e para o estudo e análise de comprometimento neurodegenerativo e outros distúrbios envolvendo distúrbios neuromusculares. The evaluation of animal locomotor activity is a behavioral tool widely used to measure the mechanisms underlying a particular disease, disorder, or injury, as well as the effects of exposure to a xenobiotic. The elevated beam test is one of the most used tests in rodents to assess balance and motor coordination. Despite being inexpensive and utilizing a simple apparatus, the high beam test requires a long period of animal training and habituation. The development and characterization of an alternative test, namely the gait test, has the potential to circumvent the time and effort required for animal training, deeming it an effective, inexpensive, and fast method for the analysis of behaviors that are comparably assessed by the high beam test. Therefore, the present study focused on determining the effectiveness and feasibility of the gait test for assessing rodent locomotion and balance as a replacement for the elevated beam test. For this purpose, male rats were divided into three groups: one control group exposed to a saline solution (NaCl 0.9%) and two experimental groups exposed to a single dose of either 0.2 or 1.0 mg/kg of ivermectin intraperitoneally for induction of locomotor disturbance. The high beam and gait tests were performed 15 min and 24 h after drug administration. Results show that the experimental groups had difficulty performing the tasks of either test at both time points analyzed compared to the control groups. At the high beam, experimental animals had trouble maintaining balance and walking. At the gait test, experimental animals showed alterations in gait, which were quantitated by: (a) shortening of step length, (b) decrease of stride, (c) altered step symmetry, and (d) altered stride area. Such results are indicative of compensatory efforts and were comparable between both tests. Altogether, the data indicate that the gait test meets all requirements for assessing motor coordination in rodents. The gait test is therefore validated as a complement to the elevated beam test for the study and analysis of neurodegenerative impairment and other disorders involving neuromuscular disturbances.

Determinação de Paraquate em maçãs do tipo Gala, Fuji, Argentina e Verde

Introduction: Brazil is one of the world leaders in consumption of agrochemicals, generating billions of reais/year and this fact is increasing. The herbicide Paraquat has a highly hydrophilic molecule and it is used in the control of weeds in many types of plantations. Objective: To determine the presence and content of Paraquat in apples using the following types from the national market: Gala, Fuji, Argentina and Verde, with or without peel and to analyze the residual Paraquat content according to the Brazilian and European legislation. Method: An adapted methodology (UV/VIS spectrophotometer) was used in apples’ types with or without peel. Results: Considering the studied types of apples, although Paraquat was present in all of them, only the Verde and Gala types would be approved by European regulations. Argentina type would be approved only by the limits stated by Anvisa and Fuji type would not be approved by none of the regulations. Conclusions: There is a need for constant inspection by analyzing the content of pesticides in food as well as legislation that establishes mandatory information about the type of agrochemical used and the corresponding residual limit allowed.Introdução: O Brasil é um dos líderes mundiais no consumo de agrotóxicos, gerando bilhões de reais por ano em valores crescentes. O herbicida Paraquate é uma molécula considerada altamente hidrofílica utilizada em plantações de milho, soja, batata, maçã, entre outros. Objetivo: Determinar a presença e o teor de Paraquate em maçãs do tipo Gala, Fuji, Argentina e Verde, em presença e ausência de casca e discutir seus limites aceitáveis nas legislações vigentes brasileira e europeia. Método: Determinar a quantidade de Paraquate por meio de metodologia adaptada utilizando um espectrofotômetro UV/VIS em amostras de maçãs já mencionadas com e sem casca. Resultados: Dos tipos estudados de maçãs considerando a fruta com a casca, embora o Paraquate estivesse presente em todas, somente os tipos Verde e Gala estariam aprovados pela regulamentação da Comissão Europeia, a Argentina seria aprovada pelos limites da Anvisa e a Fuji seria reprovada pelas duas regulamentações. Conclusões: Há a necessidade de constante fiscalização por meio da análise de teor de agrotóxicos em alimentos, bem como de novas legislações que estabeleçam a obrigatoriedade da informação do tipo de agrotóxico utilizado no produto e o limite residual permitido

Cannabinoids as a yield modifier in physical activit: A systematic review characters

Marijuana is considered illicit in much of the world, and is classified as a drug for recreational use, in recent decades the medicinal use of Cannabis sativa L. has grown and diversified, being considered the only therapeutic alternative in the control of serious and incurable diseases such as syndrome of Dravet. The world panorama has shown a more liberal position, since in several countries such as the United States of America, Holland, Australia, Italy, and more recently in Canada, the use of medications, or even the recreational use of this plant, have been regulated. In this context, the investigation of the modifying effects on physical activity of empirically used cannabinoids is fundamental nowadays, mainly due to the regulatory recognition of Cannabis sativa L. as a medicinal plant in a large part of the world. Therefore, the objective of this review was to verify the evidence related to the effect of cannabis on physical performance and to identify and highlight the challenges in the interpretation of information regarding the performance of practitioners of physical activity, as well as athletes, presenting new trends in this area of research to be addressed. To carry out the systematic review, a bibliographic survey of case reports was obtained through Pubmed, Science Direct and Google Academic databases. The following keywords were used to perform the research: cannabis, performance, pain, competition. The following filters were used as inclusion criteria a languages used english; species: humans; types of articles: original articles and reviews and period of publication of articles: 1981 to 2021

Development, Optimization, and Validation of Forensic Analytical Method for Quantification of Anticholinesterase Pesticides in Biological Matrices from Suspected Cases of Animal Poisoning

Anticholinesterase pesticides are a main cause of the intentional or accidental poisoning of animals. Anticholinesterases include several substances that cause the overstimulation of both central and peripheral acetylcholine-dependent neurotransmission. Forensic analyses of poisoning cases require high levels of expertise, are costly, and often do not provide reliable quantitative information for unambiguous conclusions. The purpose of the present study was to develop and validate a method of high-performance liquid chromatography with diode array detector (HPLC–DAD) for the identification and quantitation of n-methyl carbamates, organophosphates and respective metabolites from biological samples of animals that were suspected of poisoning. HPLC–DAD is reliable, fast, simplistic and cost-effective. The method was validated for biological samples obtained from stomach contents, liver, vitreous humor and blood from four different animal species. The validation of the method was achieved using the following analytical parameters: linearity, precision, accuracy, selectivity, recovery, and matrix effect. The method showed linearity at the range of 25–500 μg/mL, and the correlation coefficient (r2) values were >0.99 for all matrices. Precision and accuracy were determined by the (a) coefficient of variation (CV), (b) relative standard deviation low-quality control (LQC), (c) medium-quality control (QCM), and (d) high-quality control (QCA). The indicated parameters were all less than 15%. The recovery of analytes ranged from 31 to 71%. The analysis of results showed no significant interfering peaks due to common xenobiotics or matrix effects. The abovementioned method was used to positively identify pesticide analytes in 44 of the 51 animal samples that were suspected of poisoning, demonstrating its usefulness as a forensic tool

Post-Partum Depression Lactating Rat Model for Evaluating Ketamine’s Safety as a Pharmacotherapeutic Treatment: Roles in Cardiac and Urinary Function

Depression is one of the world’s most common and mentally disabling illnesses. Post-partum depression is a subtype of depression that affects one in seven women worldwide. Successful pharmacological treatment must consider the consequences for both, since the mother–child bond is fundamental for the well-being of both mother and infant as well as the general development of the newborn. Changes in maternal physiology and/or behavior can significantly influence the development of breastfed infants. Ketamine has been extensively studied for use as an antidepressant due to its mixed mechanisms of action. Safety and efficacy studies in the cardiovascular and urinary systems of a lactating postpartum depression animal model are essential for contributing toward ketamine’s clinical use in the respective patient population. Thus, this project aimed to study the implications of postpartum maternal exposure to ketamine during lactation on the cardiovascular system of female rats submitted to the depression induction model by maternal separation. This model promotes depressive effects through stress caused by the interruption of mother–infant bond early in the offspring’s life. To achieve depression, each dam was separated from her offspring for 3 h per day, from post-natal day 2 (PND2) to PND12. Experimental groups received daily treatment with either 5, 10, or 20 mg/kg of ketamine intraperitoneally during the lactation period, from PND2 to PND21. Behavioral tests consisted of the maternal and aggressive maternal behavior tests, the olfactory preference test, and the forced swim test. A technique for the detection of catecholamines and indoleamines in the heart muscle was developed for the experimental model groups. The histopathological evaluation was performed on these animals’ cardiac muscles and urinary bladders. Our findings suggest that ketamine is safe for use in postpartum depression and does not induce cardiovascular and/or urinary systems toxicity

Clinical and genetic characteristics of late-onset Huntington's disease

Background: The frequency of late-onset Huntington's disease (>59 years) is assumed to be low and the clinical course milder. However, previous literature on late-onset disease is scarce and inconclusive. Objective: Our aim is to study clinical characteristics of late-onset compared to common-onset HD patients in a large cohort of HD patients from the Registry database. Methods: Participants with late- and common-onset (30–50 years)were compared for first clinical symptoms, disease progression, CAG repeat size and family history. Participants with a missing CAG repeat size, a repeat size of ≤35 or a UHDRS motor score of ≤5 were excluded. Results: Of 6007 eligible participants, 687 had late-onset (11.4%) and 3216 (53.5%) common-onset HD. Late-onset (n = 577) had significantly more gait and balance problems as first symptom compared to common-onset (n = 2408) (P <.001). Overall motor and cognitive performance (P <.001) were worse, however only disease motor progression was slower (coefficient, −0.58; SE 0.16; P <.001) compared to the common-onset group. Repeat size was significantly lower in the late-onset (n = 40.8; SD 1.6) compared to common-onset (n = 44.4; SD 2.8) (P <.001). Fewer late-onset patients (n = 451) had a positive family history compared to common-onset (n = 2940) (P <.001). Conclusions: Late-onset patients present more frequently with gait and balance problems as first symptom, and disease progression is not milder compared to common-onset HD patients apart from motor progression. The family history is likely to be negative, which might make diagnosing HD more difficult in this population. However, the balance and gait problems might be helpful in diagnosing HD in elderly patients