Mestrado em Ciências Farmacêuticas
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- ItemAdaptação transcultural e avaliação das evidências de validade do “health care provider HIV/AIDS stigma scale” para uso no Brasil(Universidade Federal do Espírito Santo, 2024-05-03) Ribeiro, Paula Gonçalves; Torres, Thiago Silva; https://orcid.org/0000-0002-2557-601X; http://lattes.cnpq.br/6023441612547324; Araújo, Dyego Carlos Souza Anacleto de; https://orcid.org/0000-0001-6631-465X; http://lattes.cnpq.br/5120426619544250; https://orcid.org/0009-0008-1426-6279; http://lattes.cnpq.br/6360303933334244; Luz, Paula Mendes; https://orcid.org/0000-0001-9746-719X; http://lattes.cnpq.br/1664190810210313; Santos, Sabrina Cerqueira; https://orcid.org/0000-0001-6748-727X; http://lattes.cnpq.br/8757541282986479Introduction. The stigma of healthcare students towards people living with HIV (PLHIV) is a potential barrier to effective care. In Brazil, the absence of instruments assessing the stigma of healthcare students towards PLHIV prevents understanding the extent of this issue. Objective. To validate the Health Care Provider Stigma Scale (HPASS) for use among healthcare students in Brazil. Method. The research was conducted in three phases: 1) Cross-cultural adaptation of the HPASS into Brazilian Portuguese; 2) Evaluation of validity evidence based on the content of the HPASS; 3) Evaluation of the psychometric properties of the HPASS. Phase 1: Cross-cultural adaptation was performed in six steps: translation into Brazilian Portuguese, synthesis of translations, evaluation of equivalence by an expert committee, evaluation by the target audience, and back-translation into English. Phase 2: Content validity evidence was established based on feedback from a committee of four specialists. Phase 3: Psychometric properties were evaluated using a sample of Brazilian healthcare students (Nursing, Pharmacy, Medicine, Dentistry) by convenience sampling. Evidence of validity based on internal structure was examined through exploratory factor analysis (EFA). Internal consistency was assessed by composite reliability. Evidence of validity based on relationship with other variables was evaluated through correlation between HPASS scores and indices of religiosity and knowledge about HIV/aids. The project was approved by the Ethics and Research Committee of the Federal University of Espírito Santo (CAAE: 59670422.8.0000.5060 n° 5.995.451). Results. In phase 1, the cross-cultural translation and adaptation performed were considered satisfactory for the target audience and the author of the original scale. Phase 2: The content validity coefficient for the Brazilian version of the HPASS was 0.90. Phase 3: The sample consisted of 553 healthcare students. Analysis of evidence of validity based on internal structure, conducted through parallel analysis, indicated retention of two factors, with explained variance of factor 1= 42.08% (discrimination/prejudice) and factor 2= 9.92% (stereotype). Composite reliability for the factors was 0.96 (discrimination/prejudice) and 0.85 (stereotype). Evaluation of evidence of validity based on relationship with other variables showed statistically significant correlation between scores of the Brazilian version of the HPASS and religiosity and knowledge about HIV/aids. Conclusion. The Brazilian version of the HPASS is a psychometrically valid measure for assessing HIV stigma among Portuguese-speaking healthcare students in Brazil. Its psychometric properties were satisfactory, and it proved to be a simple and reliable scale. The adapted HPASS can be used for educational and research purposes among healthcare students. Future studies should investigate the scale's properties among healthcare students from different regions and healthcare professionals.
- ItemEstudo do efeito de compostos da semente de Persea americana sobre as proteínas oncogênicas VacA e CagA de Helicobacter pylori(Universidade Federal do Espírito Santo, 2025-10-28) Nunes, Amanda Cavalcante Gonzaga; Queiroz, Lorena Carnielli; https://orcid.org/0000-0002-3834-8358; http://lattes.cnpq.br/6785064719400403; Gonçalves, Rita de Cássia Ribeiro ; https://orcid.org/0000-0001-9352-2454; http://lattes.cnpq.br/6525693905417002; https://orcid.org/0009-0009-6472-778X; http://lattes.cnpq.br/8294722116448141; Silva, Ana Cândida Araújo e; https://orcid.org/0000-0002-7852-6809; http://lattes.cnpq.br/8827526324433645; Rodrigues, Ricardo Pereira ; https://orcid.org/0000-0002-2924-0468; http://lattes.cnpq.br/5051399231461377Gastric cancer is one of the most prevalent and lethal types of cancer. Its development is strongly associated with Helicobacter pylori infection. The bacterium expresses virulence factors such as cytotoxin-associated gene A (CagA) and vacuolating cytotoxin A (VacA), which induce various cellular changes, such as inhibition of pro apoptotic signaling, cell cycle dysregulation and increased release of inflammatory interleukins, characterizing them as the main oncogenic proteins of H. pylori. In this context, the present study evaluated the effect of the hydroalcoholic extract of Persea americana (avocado) seeds in vitro and its secondary metabolites in silico on the gene expression of CagA and VacA. The broth microdilution technique was used to determine the minimum inhibitory concentration, with evaluation of the minimum bactericidal concentration on blood agar. Changes in gene expression were assessed by RT-qPCR. The chemical profile of the extract was determined by high-resolution mass spectrometry. Molecular docking studies were conducted between the extract compounds and the RNA and protein structures of CagA and VacA. The extract exhibited bacteriostatic effects starting at 64 µg/mL and bactericidal effects starting at 128 µg/mL. Treatment with the extract significantly reduced cagA and vacA expression by 82.4% and 73.9% after 3h and by 76.5% and 67.1% after 6h, respectively. Fatty acid derivatives (such as avocadene and persenones), organic acids (quinic and malic), and phenolic compounds (chlorogenic acid) were proposed in the extract. Molecular docking results indicated persenones A, B, and C, and persin, as the ligands most likely to interact with the RNA structures, highlighting the most favorable complexes between domain 4 of cagA and persenone C and domain 1 of vacA and persenone A, due to a higher number of predicted hydrogen bonds. Docking with the CagA protein indicated persenone A as the most promising molecule for inhibiting the interaction between CagA and ASPP2, suggesting its potential to modulate molecular pathways associated with H. pylori-induced carcinogenesis. Predictions for the VacA protein indicated stable complexes, but the lack of consensus between programs and experimental data limits the reliability of the predictions. Based on these results, the relevance of avocado seed extract and its bioactive compounds in combating H. pylori and modulating its carcinogenic mechanisms is evident
- 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.