Teaching and learning pharmacology in Brazil before COVID-19 pandemic: a case study in Rio de Janeiro.

BACKGROUND: Knowledge of pharmacology is crucial for physicians to perform rational and safe medicine. Medical professionals are responsible for prescribing drugs and a weak performace of those can result in medication errors leading to disability, hospitalization, and death, among other situations. It occurs worldwide, including in Brazil, so that learning pharmacology impacts on public health service. We aim to investigate the current pharmacology educational practices in medical schools in the state of Rio de Janeiro, Brazil. METHODS: We surveyed 14 of 22 medical schools in Rio de Janeiro. Pharmacology teachers (n=16) and medical students (n=89) answered a semi-structured questionnaire that included questions about the staff characteristics, pharmacology content, teacher's concepts, and common practices and resources that were used in pharmacology classes. RESULTS: Our results revealed that the medical schools had similar overall curriculums. Pharmacology teachers work more than 30hs a week (75%) and conducted both research and teaching (62.5%). We also found that the multimedia projector was the most common resource (71.9%), and passive pedagogical methodologies (e.g., expository classes) remain a current strategy in pharmacology classes (89.9%). In general, medical students are poorly motivated (55%), which may be related to their performance in assessments. In addition, students believe that pharmacology is a complex (52%) or very complex subject (46%) since for its full understanding the student needs concepts from other disciplines, which can have an impact on the performance and motivation of students. As a result, these medical students do not fully understand the integration between pharmacology's basic concepts and their clinical applications. CONCLUSION: These data seem to demonstrate that the adopted teaching and learning pharmacology strategies and methodologies can be improved in Rio de Janeiro.

Open educational resources in immunology education.

The use of computers as a pedagogical resource is currently on the rise. In the case of immunology, students present difficulties in visualizing molecular phenomena. Thus the use of animations and simulations available on the internet might facilitate the learning of complex immunological concepts. In this context, it is important to map and assess the currently available resources that may be used for educational purposes. This study comprises the search and analysis of educational immunology software freely available on the internet, which can aid students and health professionals in effective learning and continuing education scenarios. A detailed search in English on the existence of free software was carried out on websites and scientific databases. The results clearly indicate a lack of freely available and scientifically validated immunology educational software, despite the existence of several software programs that could be used as auxiliary teaching tools.

Structural and molecular modeling features of P2X receptors.

Currently, adenosine 5'-triphosphate (ATP) is recognized as the extracellular messenger that acts through P2 receptors. P2 receptors are divided into two subtypes: P2Y metabotropic receptors and P2X ionotropic receptors, both of which are found in virtually all mammalian cell types studied. Due to the difficulty in studying membrane protein structures by X-ray crystallography or NMR techniques, there is little information about these structures available in the literature. Two structures of the P2X4 receptor in truncated form have been solved by crystallography. Molecular modeling has proven to be an excellent tool for studying ionotropic receptors. Recently, modeling studies carried out on P2X receptors have advanced our knowledge of the P2X receptor structure-function relationships. This review presents a brief history of ion channel structural studies and shows how modeling approaches can be used to address relevant questions about P2X receptors.

Development of Synthetic Aqueous Oil and Grease Standards for Determination of TOG in Produced Water: Evaluation of Alternative Methods for Application in an Offshore Environment.

Produced water is present in oil and natural gas reservoirs and is transported to the surface along with the oil. Total oil and grease content (TOG) is the main parameter evaluated in this waste disposal category. Today, the validation of methods in the laboratory is not done using petroleum. The objective of this work was to develop synthetic oily water standards that can be applied for internalization and validation in the laboratory. Oil weighing protocols, the influence of volatile compounds, and a procedure for preparing oily water with high reproducibility were studied. Synthetic oily water standards were prepared for TOG determination by gravimetric and infrared methods. Repeatability of 3.8 and 11% and accuracy of 85 and 105% were obtained using gravimetric and infrared methods. These results indicate that with the development of these standards, it is possible to validate methodologies for TOG determination using petroleum.

Boosting SARS-CoV-2 detection combining pooling and multiplex strategies.

RT-qPCR is the gold standard technique available for SARS-CoV-2 detection. However, the long test run time and costs associated with this type of molecular testing are a challenge in a pandemic scenario. Due to high testing demand, especially for monitoring highly vaccinated populations facing the emergence of new SARS-CoV-2 variants, strategies that allow the increase in testing capacity and cost savings are needed. We evaluated a RT-qPCR pooling strategy either as a simplex and multiplex assay, as well as performed in-silico statistical modeling analysis validated with specimen samples obtained from a mass testing program of Industry Federation of the State of Rio de Janeiro (Brazil). Although the sensitivity reduction in samples pooled with 32 individuals in a simplex assay was observed, the high-test sensitivity was maintained even when 16 and 8 samples were pooled. This data was validated with the results obtained in our mass testing program with a cost saving of 51.5% already considering the expenditures with pool sampling that were analyzed individually. We also demonstrated that the pooling approach using 4 or 8 samples tested with a triplex combination in RT-qPCR is feasible to be applied without sensitivity loss, mainly combining Nucleocapsid (N) and Envelope (E) gene targets. Our data shows that the combination of pooling in a RT-qPCR multiplex assay could strongly contribute to mass testing programs with high-cost savings and low-reagent consumption while maintaining test sensitivity. In addition, the test capacity is predicted to be considerably increased which is fundamental for the control of the virus spread in the actual pandemic scenario.

SARS-CoV-2 Variant Determination Through SNP Assays in Samples From Industry Workers From Rio de Janeiro, Brazil.

Since the first reported case in December 2019, SARS-CoV-2 infections have become a major public health worldwide. Even with the increasing vaccination in several countries and relaxing of social distancing measures, the pandemic remains a threat especially due to the emergence of new SARS-CoV-2 variants. Despite the presence of an enzyme capable of proofreading its genome, high rates of replication provide a source of accumulation of mutations within the viral genome. In this retrospective study, samples from a cohort of industry workers tested by the SESI's COVID-19 mass testing program from September 2020 to May 2021 were analyzed using a mutation panel in order to describe the circulation of currently identified SARS-CoV-2 variants within the samples obtained in Rio de Janeiro State. Our results demonstrated that the variant of interest (VOI) Zeta has been in circulation since October 2020 and reached 87% of prevalence in February 2021 followed by a decrease due to the emergence of Gamma variant of concern (VOC). Gamma was detected in January 2021 in our studied population, and its prevalence increased during the following months, reaching absolute prevalence within positive samples in May. The Alpha variant was detected only in 4-7% of samples during March and April while Beta VOC was not detected in our study. Our data agree with sequencing genomic surveillance databases and highlight the importance of continuous mass testing programs and variant detection in order to control viral spread and guide public health measures.

Developmental exposure to low-dose PBDE-99: tissue distribution and thyroid hormone levels.

Thyroid hormone concentrations, hepatic enzyme activities and tissue concentrations of 2,2',4,4',5-pentabromodiphenyl ether (PBDE-99) were evaluated in Wistar rats (dams and offspring) after treatment by gavage on gestation day (GD) 6 with a single low dose of either 60 or 300 microg PBDE-99/kg body weight (bw), respectively. Tissue concentration analysis confirmed that PBDE-99 is persistent in rodents as significant amounts of the parent compound were detected in adipose tissue 37 days after exposure. The dose of 300 microg PBDE-99/kg bw reduced thyroxin (T4) concentration in dams at the beginning of lactation (post-gestational day [PGD] 1), and caused a slight reduction in T4 on PGD 22, although not statistically significant. In offspring, reduced T4 was observed only at PND 22, probably due to cumulative effects of PBDE-99 during lactation. PBDEs have been shown to reduce T4 concentrations in several studies, but this is the first study demonstrating endocrine disruption at low doses. The adipose tissue concentration of PBDE-99 measured in this study was close to those reported for PBDE-99 in non-occupationally exposed humans. In addition, we have previously reported permanent changes in the reproductive systems and locomotor activity of male and female offspring using these same dosages.

An Improved Method for P2X7R Antagonist Screening.

ATP physiologically activates the P2X7 receptor (P2X7R), a member of the P2X ionotropic receptor family. When activated by high concentrations of ATP (i.e., at inflammation sites), this receptor is capable of forming a pore that allows molecules of up to 900 Da to pass through. This receptor is upregulated in several diseases, particularly leukemia, rheumatoid arthritis and Alzheimer's disease. A selective antagonist of this receptor could be useful in the treatment of P2X7R activation-related diseases. In the present study, we have evaluated several parameters using in vitro protocols to validate a high-throughput screening (HTS) method to identify P2X7R antagonists. We generated dose-response curves to determine the EC50 value of the known agonist ATP and the ICs50 values for the known antagonists Brilliant Blue G (BBG) and oxidized ATP (OATP). The values obtained were consistent with those found in the literature (0.7 ± 0.07 mM, 1.3-2.6 µM and 173-285 µM for ATP, BBG and OATP, respectively) [corrected].The Z-factor, an important statistical tool that can be used to validate the robustness and suitability of an HTS assay, was 0.635 for PI uptake and 0.867 for LY uptake. No inter-operator variation was observed, and the results obtained using our improved method were reproducible. Our data indicate that our assay is suitable for the selective and reliable evaluation of P2X7 activity in multiwell plates using spectrophotometry-based methodology. This method might improve the high-throughput screening of conventional chemical or natural product libraries for possible candidate P2X7R antagonist or agonist.

Virtual immunology: software for teaching basic immunology.

As immunology continues to evolve, many educational methods have found difficulty in conveying the degree of complexity inherent in its basic principles. Today, the teaching-learning process in such areas has been improved with tools such as educational software. This article introduces "Virtual Immunology," a software program available free of charge in Portuguese and English, which can be used by teachers and students in physiology, immunology, and cellular biology classes. We discuss the development of the initial two modules: "Organs and Lymphoid Tissues" and "Inflammation" and the use of interactive activities to provide microscopic and macroscopic understanding in immunology. Students, both graduate and undergraduate, were questioned along with university level professors about the quality of the software and intuitiveness of use, facility of navigation, and aesthetic organization using a Likert scale. An overwhelmingly satisfactory result was obtained with both students and immunology teachers. Programs such as "Virtual Immunology" are offering more interactive, multimedia approaches to complex scientific principles that increase student motivation, interest, and comprehension.

Effect of Rheedia longifolia leaf extract and fractions on the P2X7 receptor in vitro: novel antagonists?

Recently, the P2X(7) receptor has been reported to be associated with chronic inflammatory and neuropathic pain. Because Rheedia longifolia extract has analgesic and anti-inflammatory activity, we evaluated the in vitro inhibitory potential of methanol extract and fractions from its leaves on the P2X(7) purinergic receptor. The activity of P2X(7) was studied with a dye uptake assay and with the whole-cell patch clamp technique in mouse peritoneal macrophages treated with methanol extract of R. longifolia leaves and fractions. The dye uptake was evaluated by flow cytometry and fluorescence microscopy. The R. longifolia extract and some fractions showed an inhibitory effect on the P2X(7) purinergic receptor in a dose-dependent manner. The ethyl acetate fraction exhibited the most potent inhibitory effects. The methanol extract and the butanol fraction showed the same inhibitory effects, despite their lower potency compared with the other fractions. The R. longifolia extract and some of its fractions may be anti-inflammatory because of their inhibitory effect on the P2X(7) receptor. Further investigation is needed to determine the pattern of inhibition and selectivity. Chromatographic analysis indicated the presence of bisflavonoids in the methanol extract fractions. A member of this chemical family is the most probable active compound responsible for the P2X(7) inhibitory effects present in the R. Longifolia extract and fractions.

Desenvolvimento e avaliação de software para o ensino de ciênciasa farmacologia como modelo de interdisciplinaridade / Development and evaluation of software to teach science: pharmacology as an interdisciplinar model

As tecnologias de Informação estão cada vez mais inseridas no contexto educacional. Existe grande necessidade de um aprofundamento do conhecimento do impacto destas tecnologias na sala de aula para professores e alunos. O computador e os softwares educacionais já fazem parte integrante da realidade de muitos ambientes educacionais, contudo, mais estudos são necessários para que os benefícios do uso destas tecnologias sejam reais. Nesse contexto, se insere o ensino de Farmacologia, uma disciplina central do currículo das áreas biológicas e da saúde, importante para a formação de muitos profissionais da saúde. Dados da literatura apontam problemas no ensino desta disciplina sugerindo a necessidade de mudanças. Assim, o presente estudo consistiu de uma forma ampla, em estudar o uso das tecnologias da informação no ambiente educacional, com foco no desenvolvimento e uso de um software para o ensino da Farmacologia e o seu impacto na aprendizagem. O software foi desenvolvido sob uma perspectiva interdisciplinar, agrupando vários assuntos, com várias abordagens voltadas para a Farmacologia, uma ciência interdisciplinar por essência. Uma série de disparidades foi encontrada após uma revisão bibliográfica sobre o uso das tecnologias da informação no cenário brasileiro, assim como problemas específicos no ensino da Farmacologia nas escolas médicas do estado do Rio de Janeiro. Por outro lado, o uso do software mostrou resultados positivos sobre a aprendizagem dessa importante disciplina em estudantes de graduação das várias áreas da saúde.

Alteraçöes de isoenzimas citocromo P450 hepáticas na esquistossomose mansônica murina / Alterations of isoenzimas citocromo P450 liverworts in esquistossomose mansônica murina

Tem sido relatado que infecçöes e estímulos inflamatórios causam alteraçöes da atividade e dos níveis de expressäo de várias formas de citocromos P450 (CYP) hepáticos em animais de laboratório e em seres humanos. Estas alteraçöes podem ter reflexos tanto no clearance de fármacos quanto na ativaçäo metabólica de pró-mutágenos, pró-carcinógenos e pró-teratógenos. Assim, infecçöes e processos inflamatórios estäo entre os fatores que modulam a atividade ou a expressäo dos CYP e podem alterar a cinética e a toxicidade de xenobióticos. Teve como objetivo avaliar os efeitos da infecçäo com S., mansoni sobre o conteúdo total de citocromos P450, as atividades de isoenzimas das subfamílias 1A, 2A e 2B, e os níveis de CYP10 e 2A5 na fraçäo microssomal hepática de camundongos das linhagens Swiss Webster e DBA/2. Avalia a evoluçäo da esquistossomose murina em Swiss Webster e DBA/2 comparando dois modelos de infecçäo experimental animais infectados com 100 cercárias aos 10 dias de vida (lactentes) ou na vida adulta (45 dias de vida). Foram determinados o peso corporal e de órgäos (fígado, baço e intestinados), o ganho ponderal, o índice de penetraçäo das cercárias, a mortalidade, a taxa de recuperaçäo de vermes após perfusäo sistêmica e o número de ovos no fígado e intestinos. Indica que na fase intermediária de evoluçäo da esquistossomose mansônica murina (55 dias pós infecçäo) há uma acentuada diminuiçäo dos níveis totais de CYP e de atividades relacionadas as subfamílias CYP1A e CYP2B. Por outro lado, contrastando com esta depressäo generalizada de CYPs, há um claro aumento de CoH, relacionada ao CYP2A5/2 infectados.