Mestrado em Ciências Farmacêuticas
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- ItemA anestesia com ketamina/xilazina altera a atividade colinesterásica em ratos: uma abordagem in vivo, in vitro e in silico(Universidade Federal do Espírito Santo, 2025-10-03) Corrêa, Larissa de Jesus; Gonçalves, Juliana Barbosa Coitinho ; https://orcid.org/0000-0002-5892-050X; http://lattes.cnpq.br/3448669742301744; Sampaio, Karla Nívea; https://orcid.org/0000-0003-0293-0482; http://lattes.cnpq.br/5951704470576361; https://orcid.org0009-0007-2152-3432; http://lattes.cnpq.br/3563882736771738; Dias Júnior, Carlos Alan Candido ; https://orcid.org/0000-0002-0348-6144; http://lattes.cnpq.br/6296664642422599; Gonçalves, Rita de Cássia Ribeiro; https://orcid.org/0000-0001-9352-2454; http://lattes.cnpq.br/6525693905417002Introduction: Ketamine and Xylazine (K/X) are veterinary anesthetics commonly used in various procedures, but they can interfere with the body's homeostasis and autonomic balance. Previous studies have observed a reduction in plasma butyrylcholinesterase (BChE) activity with K/X, suggesting possible systemic cholinergic modulation. However, it remains unknown whether this effect extends to other cholinesterases or to different tissues, as well as the possible mechanisms involved in these effects. To elucidate these questions, we integrated in silico, in vitro, and in vivo approaches to investigate the interaction of K/X with cholinesterases and evaluate its effects on enzyme activity and the expression of inflammatory markers. Methods: The in silico study investigated the binding potential of K/X to cholinesterases and predicted interactions with catalytic, anionic, and peripheral residues. In vitro assays evaluated the dose-dependent effect of each drug on erythrocyte acetylcholinesterase (AChE) activity and plasma BChE in control rats. In the in vivo study, Wistar rats (10–12 weeks) were assigned to four protocols: (1) serial blood collection before, during, and after K/X anesthesia and surgery; (2) isolated anesthesia followed by euthanasia 48 h later; (3) control group without prior exposure to anesthesia/surgery; and (4) caudal puncture without anesthesia and euthanasia three days later. Finally, blood, cortex, hippocampus, brainstem, and heart samples were collected for analysis of AChE and BChE, as well as Western blot analysis of TNF-α and NF-κB expression in the left ventricle (LV). Results: Docking indicated weak interactions between cholinesterase residues and ligands. In vitro testing showed dose-dependent inhibition of AChE and BChE in control rats by both anesthetics. Surgery and anesthesia combined reduced plasma BChE activity 24 and 48 hours after anesthesia, and erythrocyte AChE activity 48 hours after anesthesia. Anesthesia alone decreased BChE activity but increased AChE activity. The caudal puncture did not change enzyme activity. Anesthesia with or without surgery increased atrial cholinesterase activity and decreased ChE activity in the left ventricle, brainstem, and prefrontal cortex. TNF-α expression in the LV was decreased by K/X. Conclusions: Anesthetic induction with K/X changes cholinesterase activity in a tissue- and time dependent manner, suggesting a systemic effect on cholinergic tone. The concomitant reduction of TNF-α in the LV indicates a possible link between cholinergic modulation and the inflammatory response.
- ItemEfeito da exposição aguda ao clorpirifós nas respostas autonômicas e comportamentais em ratos no medo condicionado ao contexto: a influência de diferentes condições experimentais no desfecho(Universidade Federal do Espírito Santo, 2025-08-28) Noé, Gabriel Gavazza; Harres, Vanessa Beijamini; https://orcid.org/0000-0003-4533-0495; http://lattes.cnpq.br/5077271160260796; Sampaio, Karla Nívea; https://orcid.org/0000-0003-0293-0482; http://lattes.cnpq.br/5951704470576361; https://orcid.org/0009-0005-6857-6983; http://lattes.cnpq.br/8529914893110844 ; Emerick, Guilherme Luz; https://orcid.org/0000-0002-6206-3985; http://lattes.cnpq.br/5067407796070598; Carlétti, Isis Moraes Ornelas; https://orcid.org/0000-0002-8765-3324; http://lattes.cnpq.br/4871859538334419; 3º membro da banca; https://orcid.org; https://lattes.cnpq.brOrganophosphates (OPs) are a class of compounds widely used worldwide. Evidence suggests that different exposure patterns to OP compounds induce cardiovascular, respiratory, behavioral, cognitive, motor, oxidative/nitrosative, and inflammatory impairments. In previous studies, we observed that a single-dose intoxication with chlorpyrifos (CPF), an OP, caused behavioral impairments in contextual fear conditioning (CFC). However, we did not evaluate whether acute intoxication would impair the autonomic response in CFC. Furthermore, studies show that the use of anesthetics, such as ketamine and xylazine, can interfere with behavioral and cardiovascular responses. Therefore, we investigated the behavioral, autonomic, and biochemical effects of acute exposure to CPF in rats subjected to CFC, exploring how variations in experimental conditions, such as protocol order, anesthesia, surgery, and time, can influence the results obtained. To this end, adult Wistar rats were subjected to the CFC test. The CFC was divided into three sessions: conditioning, extinction, and recall extinction. Exposure to CPF (20 mg/kg, i.p.) always occurred after conditioning. Autonomic (mean arterial pressure, MAP, and heart rate, HR) and behavioral (freezing) parameters were recorded during the extinction and recall extinction sessions. Four different protocols were performed: Protocol 1 – femoral artery cannulation surgery under ketamine/xylazine (80/10 mg/kg) anesthesia 24 h before conditioning and the effect of CPF on the CFC 24 h after intoxication; Protocol 2 – cannulation surgery 24 h after conditioning and the effect of CPF on the CFC 48 h after intoxication; Protocol 3 – extinction session 48 h after conditioning and CPF administration, in the absence of surgery and anesthesia; Protocol 4 – anesthesia with the ketamine/xylazine mixture, 24 h after conditioning, in the absence of surgery. At the end of the experiments, the animals were euthanized for sample collection to assess acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) enzyme activity, and to quantify nitrosative stress markers (NO2-, NO3-, and total NO) in the hippocampus and trunk. In protocol 1, acute CPF intoxication impaired extinction and increased fear expression 24 h after intoxication without affecting MAP or HR. Conversely, in protocol 2, acute CPF exposure altered the pressor and tachycardic responses of intoxicated animals that underwent surgery 24 h after conditioning, without affecting behavioral responses. In protocols 3 and 4, acute CPF poisoning impaired fear extinction, while the anesthetic combination of ketamine and xylazine and the anesthetic combination with CPF also impaired the extinction of conditioned fear. In all four protocols, it was observed that poisoning led to cholinesterase inhibition. Furthermore, acute CPF poisoning altered the levels of NO2-, NO3-, and total NO in animals 72 h after poisoning. Thus, it is concluded that acute CPF poisoning of rats impairs behavioral and cardiovascular responses in the CFC, and that the anesthetic combination and surgery may interfere with these results.
- ItemEfeito da exposição aguda ao clorpirifós nas respostas autonômicas e comportamentais em ratos no medo condicionado ao contexto: a influência de diferentes condições experimentais no desfecho(Universidade Federal do Espírito Santo, 2025-08-28) Noé, Gabriel Gavazza; Harres, Vanessa Beijamini; https://orcid.org/0000-0003-4533-0495; http://lattes.cnpq.br/5077271160260796; Sampaio, Karla Nívea; https://orcid.org/0000-0003-0293-0482; http://lattes.cnpq.br/5951704470576361; https://orcid.org/0009-0005-6857-6983; http://lattes.cnpq.br/8529914893110844 ; Emerick, Guilherme Luz; https://orcid.org/0000-0002-6206-3985; http://lattes.cnpq.br/5067407796070598; Carletti, Isis Moraes Ornelas; https://orcid.org/0000-0002-8765-3324; http://lattes.cnpq.br/4871859538334419Organophosphates (OPs) are a class of compounds widely used worldwide. Evidence suggests that different exposure patterns to OP compounds induce cardiovascular, respiratory, behavioral, cognitive, motor, oxidative/nitrosative, and inflammatory impairments. In previous studies, we observed that a single-dose intoxication with chlorpyrifos (CPF), an OP, caused behavioral impairments in contextual fear conditioning (CFC). However, we did not evaluate whether acute intoxication would impair the autonomic response in CFC. Furthermore, studies show that the use of anesthetics, such as ketamine and xylazine, can interfere with behavioral and cardiovascular responses. Therefore, we investigated the behavioral, autonomic, and biochemical effects of acute exposure to CPF in rats subjected to CFC, exploring how variations in experimental conditions, such as protocol order, anesthesia, surgery, and time, can influence the results obtained. To this end, adult Wistar rats were subjected to the CFC test. The CFC was divided into three sessions: conditioning, extinction, and recall extinction. Exposure to CPF (20 mg/kg, i.p.) always occurred after conditioning. Autonomic (mean arterial pressure, MAP, and heart rate, HR) and behavioral (freezing) parameters were recorded during the extinction and recall extinction sessions. Four different protocols were performed: Protocol 1 – femoral artery cannulation surgery under ketamine/xylazine (80/10 mg/kg) anesthesia 24 h before conditioning and the effect of CPF on the CFC 24 h after intoxication; Protocol 2 – cannulation surgery 24 h after conditioning and the effect of CPF on the CFC 48 h after intoxication; Protocol 3 – extinction session 48 h after conditioning and CPF administration, in the absence of surgery and anesthesia; Protocol 4 – anesthesia with the ketamine/xylazine mixture, 24 h after conditioning, in the absence of surgery. At the end of the experiments, the animals were euthanized for sample collection to assess acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) enzyme activity, and to quantify nitrosative stress markers (NO2-, NO3-, and total NO) in the hippocampus and trunk. In protocol 1, acute CPF intoxication impaired extinction and increased fear expression 24 h after intoxication without affecting MAP or HR. Conversely, in protocol 2, acute CPF exposure altered the pressor and tachycardic responses of intoxicated animals that underwent surgery 24 h after conditioning, without affecting behavioral responses. In protocols 3 and 4, acute CPF poisoning impaired fear extinction, while the anesthetic combination of ketamine and xylazine and the anesthetic combination with CPF also impaired the extinction of conditioned fear. In all four protocols, it was observed that poisoning led to cholinesterase inhibition. Furthermore, acute CPF poisoning altered the levels of NO2-, NO3-, and total NO in animals 72 h after poisoning. Thus, it is concluded that acute CPF poisoning of rats impairs behavioral and cardiovascular responses in the CFC, and that the anesthetic combination and surgery may interfere with these results.
- ItemAvaliação do conhecimento de farmacêuticos e estudantes de farmácia sobre o transtorno do espectro autista no Brasil(Universidade Federal do Espírito Santo, 2024-04-05) Rosa, Larissa Couto; Araújo, Dyego Carlos Souza Anacleto de; https://orcid.org/0000-0001-6631-465X; http://lattes.cnpq.br/5120426619544250; Santos Júnior, Genival Araújo dos; https://orcid.org/0000-0002-5618-1846; http://lattes.cnpq.br/5917097187533724; http://lattes.cnpq.br/2475269525071153; Rocha, Kérilin Stancine Santos; https://orcid.org/0000-0002-2313-2140; http://lattes.cnpq.br/4216594263272412; Silva, Rafaella de Oliveira Santos; http://lattes.cnpq.br/4008596465243842Pharmacy students’ and pharmacists’ limited knowledge about Autism Spectrum Disorder (ASD) can have negative impacts on health care for these individuals. In Brazil, there are no studies that assess the knowledge of Pharmacy students and pharmacists about ASD. Therefore, the objective of this study is to assess the knowledge of Pharmacy students and pharmacists about ASD in Brazil. A cross-sectional survey study was conducted between September 2021 and August 2022. Pharmacy students and pharmacists from all five regions of Brazil participated in the survey. Data were collected through an online questionnaire that contained two sections: the first addressed sociodemographic data and contact with people with ASD and the second section contained the items that make up the Autism Stigma Knowledge – Questionnaire (ASK-Q) Brazil. This questionnaire assesses knowledge and stigma about ASD in 49 questions divided into 4 subscales. The instrument considers the following scores for adequate knowledge: diagnosis (11-18 points); etiology (11-16 points); treatment (10-14 points); non-endorsement of stigma (3-7 points). Descriptive statistics, t test to compare mean scores, and Mann-Whitney test to compare unpaired samples were used for statistical analyses. This study was approved by the Research Ethics Committee (Opinion No. 4,464,411). A total of 397 pharmacy students and 413 pharmacists participated in the study. Most students had adequate knowledge in the following domains: etiology (99.0%), diagnosis (97.0%), and treatment (98.7%), and no student demonstrated endorsement of stigma (100.0%). The highest rates of correct answers in the total score were observed among students from public institutions (p=0.001; d=0.5). In the treatment domain, the highest scores were observed among students who had a family member with ASD (p=0.001; d=0.45) and who studied in public institutions (p=0.002; d=0.43). In the diagnosis domain, the highest scores were observed among those who had a family member with ASD (p=0.015; d=0.24) and lived with a person with ASD (p=0.007; d=0.3). Most pharmacists presented adequate knowledge in the domains: etiology (99.0%), diagnosis (99.0%), treatment (98.8%), and no pharmacist demonstrated to endorse stigma (100.0%). In the diagnosis domain, the highest scores were observed among pharmacists who reported having a family member with ASD (p<0.001) and living with a person with ASD (p=0.002). In the etiology domain, the highest scores were among people who lived with a person with ASD (p=0.004). This study revealed that the pharmacists and pharmacy students analyzed had adequate knowledge about ASD. The results may contribute to the development of strategies that improve the continuing education of Brazilian pharmacists.
- ItemPesquisa de bactérias resistentes aos antimicrobianos em duas localidades do Estado do Espírito Santo(Universidade Federal do Espírito Santo, 2025-02-27) Mothé, Davi Abreu Carvalho; Santos, Kênia Valéria dos; https://orcid.org/0000-0001-6871-3128; http://lattes.cnpq.br/9074173162086323; http://lattes.cnpq.br/8506942685475849; Apolônio, Ana Carolina Morais; https://orcid.org/0000-0001-9049-7660; http://lattes.cnpq.br/7570808562726658; Resende, Juliana Alves; https://orcid.org/0000-0002-5476-3754; http://lattes.cnpq.br/8223821041049149Antimicrobial resistance (AMR) is a growing concern, especially due to the spread of resistant bacteria in the environment, which can act as a reservoir for resistance genes. Vitória Bay and the Caramuru district in Espírito Santo, Brazil, stand out as hotspots for the study of AMR. The former because of the intense port activity and high flow of people and goods, and the latter because of agricultural and poultry production with intensive use of antimicrobials. Selective antimicrobial media were used to screen for the presence of antimicrobial-resistant bacteria in water samples from Vitória Bay and in environmental samples from the Caramuru district: water, soil, poultry litter, wet bathroom surfaces, animal feces and flies. In Caramuru, 138 isolates were identified with a predominance of Escherichia coli and Enterobacter, while in Vitória Bay, 50 isolates were identified with a predominance of pathogens such as Klebsiella pneumoniae and Acinetobacter pittii. Among the Gram-positive cocci, only Enterococcus spp. were found, one of which was resistant to vancomycin in Vitória Bay. Phenotypic tests confirmed the presence of multidrug-resistant isolates in different environmental sources, with extended-spectrum beta-lactamase (ESBL) production more frequent in Vitória Bay. The production of metallo-beta-lactamases was observed in Enterobacter spp. in Caramuru and in K. pneumoniae in Vitória Bay, while resistance to polymyxin B was observed in E. asburiae, E. cloacae, E. coli, A. baumannii and P. aeruginosa isolated from soil, toilets and water. Vitória Bay showed higher frequencies of MDR bacteria and resistance genes, including blaKPC, qnrS, rmtA and rmtG, while in Caramuru only blaTEM and blaCTX-M-1/2 were detected. These results show the diversity and prevalence of resistant bacteria in both rural and urban areas. These results demonstrate the diversity and prevalence of resistant bacteria in rural and urban areas. The detection of MDR bacteria in environments close to human interaction, including water used for irrigation and coastal areas, highlights a significant risk for the spread of antimicrobial resistance. These findings emphasize the urgent need for surveillance and control.