The tissue-specific nature of physiological zebrafish mitochondrial bioenergetics.

Zebrafish are a powerful tool to study a myriad of experimental conditions, including mitochondrial bioenergetics. Considering that mitochondria are different in many aspects depending on the tissue evaluated, in the zebrafish model there is still a lack of this investigation. Especially for juvenile zebrafish. In the present study, we examined whether different tissues from zebrafish juveniles show mitochondrial density- and tissue-specificity comparing brain, liver, heart, and skeletal muscle (SM). The liver and brain complex IV showed the highest O2 consumption of all ETC in all tissues (10x when compared to other respiratory complexes). The liver showed a higher potential for ROS generation. In this way, the brain and liver showed more susceptibility to O2- generation when compared to other tissues. Regarding Ca2+ transport, the brain showed greater capacity for Ca2+ uptake and the liver presented low Ca2+ uptake capacity. The liver and brain were more susceptible to producing NO. The enzymes SOD and Catalase showed high activity in the brain, whereas GPx showed higher activity in the liver and CS in the SM. TEM reveals, as expected, a physiological diverse mitochondrial morphology. The essential differences between zebrafish tissues investigated probably reflect how the mitochondria play a diverse role in systemic homeostasis. This feature may not be limited to normal metabolic functions but also to stress conditions. In summary, mitochondrial bioenergetics in zebrafish juvenile permeabilized tissues showed a tissue-specificity and a useful tool to investigate conditions of redox system imbalance, mainly in the liver and brain.

Collagen content and C-X-C motif chemokine ligand 12 expression in neoplastic breast stroma.

OBJECTIVE: This study aimed to evaluate the expression of C-X-C motif chemokine ligand 12 and its C-X-C chemokine receptor type 4, and the tumor-stroma ratio using collagen stromal content of breast cancer samples, correlating it with clinicopathological data. METHODS: Through a retrospective cohort study, samples were obtained from female patients, over 18 years of age, with the disease in stages 1-4, who underwent mastectomy or lumpectomy. The biopsies were provided by the Oncology sector of the Hospital das Clínicas of Universidade Federal de Pernambuco, Recife city, in 2011-2014, including samples of invasive ductal carcinoma, ductal carcinoma in situ, or benign changes (fibroadenoma and hypertrophy), which were analyzed between 2020 and 2022 by immunohistochemistry for the expression of stromal cell characteristics. Collagen content was tested by Gomori staining and digital analysis of images. RESULTS: Absence of stromal expression of C-X-C motif chemokine ligand 12 was associated with longer disease-free survival (disease-free survival=0.481), and expression of C-X-C chemokine receptor type 4 was associated with lower disease-free survival. An association was observed between clinicopathological variables and stromal expression of chemokines, that is, an association of stromal C-X-C motif chemokine ligand 12 with histological grade, angiolymphatic invasion, and an association between C-X-C chemokine receptor type 4 expression and histological grade. Analyses of digital pixels images of collagen and tumor cells showed a lower percentage of collagen in the invasive ductal carcinoma samples (39%), unlike samples without neoplasms (78%). CONCLUSION: Low expression of C-X-C motif chemokine ligand 12 may be associated with a worse prognosis for breast cancer. Collagen staining analyzed using digital images represents an opportunity for clinical application and is indicative of the prognosis of the tumor microenvironment in breast carcinoma.

Collagen content and C-X-C motif chemokine ligand 12 expression in neoplastic breast stroma

SUMMARY OBJECTIVE: This study aimed to evaluate the expression of C-X-C motif chemokine ligand 12 and its C-X-C chemokine receptor type 4, and the tumor-stroma ratio using collagen stromal content of breast cancer samples, correlating it with clinicopathological data. METHODS: Through a retrospective cohort study, samples were obtained from female patients, over 18 years of age, with the disease in stages 1-4, who underwent mastectomy or lumpectomy. The biopsies were provided by the Oncology sector of the Hospital das Clínicas of Universidade Federal de Pernambuco, Recife city, in 2011-2014, including samples of invasive ductal carcinoma, ductal carcinoma in situ, or benign changes (fibroadenoma and hypertrophy), which were analyzed between 2020 and 2022 by immunohistochemistry for the expression of stromal cell characteristics. Collagen content was tested by Gomori staining and digital analysis of images. RESULTS: Absence of stromal expression of C-X-C motif chemokine ligand 12 was associated with longer disease-free survival (disease-free survival=0.481), and expression of C-X-C chemokine receptor type 4 was associated with lower disease-free survival. An association was observed between clinicopathological variables and stromal expression of chemokines, that is, an association of stromal C-X-C motif chemokine ligand 12 with histological grade, angiolymphatic invasion, and an association between C-X-C chemokine receptor type 4 expression and histological grade. Analyses of digital pixels images of collagen and tumor cells showed a lower percentage of collagen in the invasive ductal carcinoma samples (39%), unlike samples without neoplasms (78%). CONCLUSION: Low expression of C-X-C motif chemokine ligand 12 may be associated with a worse prognosis for breast cancer. Collagen staining analyzed using digital images represents an opportunity for clinical application and is indicative of the prognosis of the tumor microenvironment in breast carcinoma.

Effects of risk factors in the likelihood of breast cancer subtypes in Brazilian women / Efeitos de fatores de risco na probabilidade de subtipos do câncer de mama em mulheres brasileiras

INTRODUCTION: Breast cancer is a heterogenous disease with multiple causes and it lacks more investigation related to its risk factors. OBJECTIVE: To evaluate the likelihood of breast cancer subtypes according to the positivity to estrogen and progesterone receptors (ER+ and PR+ respectively), with or without the expression HER2, related to the following risk factors: age, parity, diabetes mellitus, arterial hypertension, occurrence of familiar cancer case and body mass index (BMI). METHODS: The sample with 79 individuals was divided into three subtypes 1 (ER+/PR-), 2 (ER+/PR+) and 3 (RE+/RP+/HER+) and then analyzed by quantitative methods using Ordinal Generalized Linear Models (OGLM) for estimating the marginal effects of risk factors for the studied subtypes, and modeling the heteroscedasticity in terms of error. RESULTS: It were observed the following statistically significant positive effects: (1) age for the tumoral subtype 1 (ER+/PR-) and (2) parity for the subtype 2 (ER+/PR+); while the significant negative effects were: (1) age for subtype 3 (ER+/PR+/HER2+); (2) parity for both 1 (ER+/PR-) and 3 (ER+/PR+/HER2+) subtypes; and arterial hypertension for subtype 1 (ER+/PR-). There were no statistically significant effects for BMI, Diabetes mellitus and occurrence of familiar cancer variables on the studied tumoral subtypes. CONCLUSION: The risk factos age and parity demonstrated varied effects for the breast cancer subtypes according the expression of estrogen, progesterone and HER2 receptors.

INTRODUÇÃO: O câncer de mama é uma doença heterogênea, multifatorial e que necessita de mais estudos relacionados aos fatores de riscos. OBJETIVO: Analisar a probabilidade dos subtipos tumorais do câncer de mama receptores de estrogênio (RE) ou progesterona (RP) positivos, com ou sem expressão de HER2, em relação aos fatores de risco: idade, parto, diabetes mellitus, hipertensão arterial, ocorrência de câncer de mama familiar e índice de massa corporal (IMC). MÉTODOS: A análise da amostra de 79 pacientes dividida nos subtipos 1 (RE+/RP-), 2 (RE+/RP+) e 3 (RE+/RP+/HER+), foi feita por meio de métodos quantitativos usando o Modelo Linear Generalizado Ordinal (MLGO) para estimar os efeitos marginais dos fatores de risco para os subtipos estudados, e ao mesmo tempo modelando a heterocedasticidade do termo de erro. RESULTADOS: Nos resultados foram observados os seguintes efeitos positivos estatisticamente significantes: (1) da idade para o subtipo tumoral 1 (RE+/RP-) e (2) do número de partos para o subtipo 2 (RE+/RP+); enquanto os efeitos negativos significativos foram os seguintes: (1) da idade para o subtipo 3 (RE+/RP+/HER2+); (2) do número de partos para o sutipos 1 (RE+/RP) e 3 (RE+/RP+/HER2+); e da hipertensão arterial para o o subtipo 1 (RE+/RP-). Não foram observados efeitos estatisticamente significativos das variáveis IMC, Diabetes mellitus e ocorrência de câncer de mama familiar sobre os subtipos tumorais estudados. CONCLUSÃO: Os fatores de risco idade e número de partos têm efeitos variados para os subtipos do câncer de mama segundo a expressão de receptores para estrogênio, progesterona e HER2.

Novel 4-quinoline-thiosemicarbazone derivatives: Synthesis, antiproliferative activity, in vitro and in silico biomacromolecule interaction studies and topoisomerase inhibition.

Twelve 2-(quinolin-4-ylmethylene) hydrazinecarbothioamide derivatives were synthetized and their biological properties were investigated, among which, the ability to interact with DNA and BSA through UV-Vis absorption, fluorescence, Circular Dichroism, molecular docking and relative viscosity, antiproliferative activity against MCF-7 and T-47D mammary tumor cells and RAW-264.7 macrophages and inhibitory capacity of the enzyme topoisomerase IIα. In the binding study with DNA and BSA, all the compounds displayed affinity for interaction with both biomolecules, especially JF-92 (p-ethyl-substituted), with binding constant of 1.62 × 106 and 1.43 × 105, respectively, and DNA binding mode by intercalation. The IC50 values were obtained between 0.81 and 1.48 µM and topoisomerase inhibition results in 10 µM. Thus, we conclude that the reduction of the acridine to quinoline ring did not disrupt the antitumor action and that substitution patterns are important for biomolecule interaction affinity as they demonstrate the potential of these compounds for anticancer therapy.

Curricularização da Extensão Universitária no Ensino Médico: o Encontro das Gerações para Humanização da Formação / Curricularisation of University Community Outreach in Medical Education: Meeting of Generations for a Humanized Training

RESUMO Este trabalho relata uma experiência de curricularização da extensão universitária no curso médico que teve como objetivos conduzir ações de extensão com vistas ao bem-estar de idosas de uma Instituição de Longa Permanência (ILP), bem como desenvolver habilidades do médico honesto, íntegro, cuidadoso, altruísta e empático consigo, com os "idosos" e com os pares envolvidos nas ações. O modelo usado para organização das atividades combinou métodos de aprendizagem de habilidades por meio da experiência, reflexão crítica sobre as experiências, ambiente de pequeno grupo para suporte e validação e programa longitudinal coeso para desenvolvimento total. As atividades foram vivenciadas em grupos de 12 integrantes, supervisionados por um professor com visitas semanais à ILP de agosto a dezembro de 2018. O modelo utilizado mostrou-se útil para o desenvolvimento de características humanísticas almejadas nos indivíduos em formação. O envolvimento em todas as etapas do processo e o comprometimento com o bem-estar da população selecionada indicam que a curricularização cumpriu seu papel de proporcionar aos estudantes o desenvolvimento de consciência cidadã em paralelo à formação técnica. A instituição de ensino superior deve oferecer continuamente ações para que os estudantes desenvolvam plenamente seu lado humano e preservem as características observadas no contato com a população idosa.

ABSTRACT This work describes the experience of including community outreach activities of a university into the medical course curriculum, with the objectives of developing outreach actions directed at the well-being of the elderly from a Long Stay Institution (ILP). At the same time the work promoted the development of honest, caring, selfless professionals empathetic toward the "elderly" and peers involved in the actions. The model used to organize the activities combined methods of learning skills through experience, critical reflection on experiences, small group environment for support and validation, and a cohesive longitudinal program for total development. The activities were experienced in groups of 12 supervised by a teacher with weekly visits to the ILP from August to December of 2018. The model used proved to be useful for the development of humanistic characteristics. Involvement in all stages of the process and commitment to the well-being of the selected population indicate that the curricularisation fulfilled its role of providing students with the development of citizen awareness and technical training. The institution must continually offer actions for students to fully develop their human side and preserve the characteristics observed in their contact with the elderly population.

Synthesis, in vitro and in vivo biological evaluation, COX-1/2 inhibition and molecular docking study of indole-N-acylhydrazone derivatives.

The objective of this work was to obtain and evaluate anti-inflammatory in vitro, in vivo and in silico potential of novel indole-N-acylhydrazone derivatives. In total, 10 new compounds (3a-j) were synthesized in satisfactory yields, through a condensation reaction in a single synthesis step. In the lymphoproliferation assay, using mice splenocytes, 3a and 3b showed inhibition of lymphocyte proliferation of 62.7% (±3.5) and 50.7% (±2), respectively, while dexamethasone presented an inhibition of 74.6% (±2.4). Moreover, compound 3b induced higher Th2 cytokines production in mice splenocytes cultures. The results for COX inhibition assays showed that compound 3b is a selective COX-2 inhibitor, but with less potency when compared to celecoxib, and compound 3a not presented selectivity towards COX-2. The molecular docking results suggest compounds 3a and 3b interact with the active site of COX-2 in similar conformations, but not with the active site of COX-1, and this may be the main reason to the COX-2 selectivity of compound 3b. In vivo carrageenan-induced paw edema assays were adopted for the confirmation of the anti-inflammatory activity. Compound 3b showed better results in suppressing edema at all tested concentrations and was able to induce an edema inhibition of 100% after 5 h of carrageenan injection at the 30 mg kg-1 dosage, corroborating with the COX inhibition and lymphoproliferation results. I addition to our experimental results, in silico analysis suggest that compounds 3a and 3b present a well-balanced profile between pharmacodynamics and pharmacokinetics. Thus, our preliminary results revealed the potentiality of a new COX-2 selective derivative in the modulation of the inflammatory process.

Peer-Assisted and Team-Based Learning: A new hybrid strategy for Medical Education / Monitoria e Aprendizagem Baseada em Equipes: Uma nova estratégia híbrida para Educação Médica

ABSTRACT Medical education has been significantly changing in the last years, mainly due to new conceptions of the teaching-learning processes that advocate the employment of active methods of learning, such as Team-Based Learning (TBL). Peer-Assisted Learning (PAL) is a long-established strategy that has been used in several undergraduate courses. This study describes a pilot test of a new model of PAL developed with an adapted TBL strategy, that was called "PAL-TBL method". The test occurred during Human Physiology classes with first-year Medical undergraduate students from Universidade de Pernambuco - Garanhuns Campus, Brazil. The PAL-TBL was designed as a teaching-learning methodology to improve academic education. Here, the organizational dynamics and the design of the activities carried out from 2016 to 2017 are reported. The resulting PAL-TBL methodology is characterized as the following: (i) timing I or material preparation (context/scenario) and study/analysis of the material by the participants; (ii) timing II or verification of prior knowledge (individual and team test), questioning and feedback and (iii) timing III or applying the concepts learned. It is worth mentioning that the end of timing II consisted of a moment for evaluating the team's work and the materials used. Material production happened through the interaction between student-tutors and the (supervising) professor, aiming to share experiences as well as to elucidate the importance of using active methods during the academic development of useful competencies for medical practice. The methodology developed allowed students to reflect extensively on the problems presented in the class discussions, allowing for a richer learning experience. Further applications of PAL-TBL in advanced years of the course will be done to confirm the benefits of this hybrid strategy for medical education.

RESUMO A educação médica tem mudado bastante nos últimos anos, principalmente pelo surgimento de novos conceitos relacionados aos processos de ensino-aprendizagem que preconizam o emprego de métodos ativos, tais como a Aprendizagem Baseada em Equipes (ABE), do inglês Team-Based Learning (TBL). A monitoria é uma estratégia bem estabelecida para atividades de ensino que tem sido utilizada em vários cursos de graduação. Esse artigo descreve o teste piloto de um novo modelo de monitoria desenvolvido por meio da adaptação do ABE, então denominado método "PAL-TBL". O método foi aplicado durante as aulas de Fisiologia Humana com os estudantes do primeiro ano do curso de Medicina da Universidade de Pernambuco - campus Garanhuns, Brasil. O método PAL-TBL foi delineado como uma modalidade de ensino-aprendizagem para contribuir para a formação acadêmica dos estudantes de Medicina. Nesse sentido, a dinâmica organizacional e o desenvolvimento das atividades deram-se durante os anos de 2016 e 2017. A metodologia utilizada foi a seguinte: (i) momento I ou de preparação de material (contexto/cenário) e estudo/análise desse material pelos participantes; (ii) momento II de verificação do conhecimento prévio (teste individual e em equipe), levantamento de dúvidas e feedback e (iii) momento III de aplicação dos conceitos. Vale ressaltar que o momento II contou com momento avaliativo do trabalho das equipes e do material preparado. A produção do material, contexto, questões e devolutiva, foi realizada por meio da interação entre os estudantes monitores e o professor, com o intuito de inserir as experiências vividas previamente, bem como elucidar a importância do uso de métodos ativos durante a formação acadêmica para o desenvolvimento de competências úteis para a prática médica. O desenvolvimento da metodologia proposta levou os estudantes a refletirem mais sobre os problemas apresentados durante as discussões em sala de aula, permitindo uma rica experiência de aprendizagem. Aplicações futuras do PAL-TBL com estudantes de anos mais avançados do curso serão realizadas para ampliar os benefícios da estratégia desenvolvida para a formação médica.

New spiro-acridines: DNA interaction, antiproliferative activity and inhibition of human DNA topoisomerases.

Two new spiro-acridines were synthesized by introducing cyano-N-acylhydrazone between the acridine and phenyl rings followed by spontaneous cyclization. The final compounds (E)-1'-(benzylideneamino)-5'-oxo-1',5'-dihydro-10H-spiro[acridine-9,2'-pyrrole]-4'-carbonitrile (AMTAC-01) and (E)-1'-((4-methoxybenzylidene)amino)-5'-oxo-1',5'-dihydro-10H-spiro[acridine-9,2'-pyrrole]-4'-carbonitrile (AMTAC-02) were evaluated for their interactions with calf thymus DNA, antiproliferative and human topoisomerase I and IIα inhibitory activities. Both compounds presented ability to bind DNA. The binding constant determined by UV-vis spectroscopy was found to be 104M-1. Antiproliferative assay demonstrated that AMTAC-01 and AMTAC-02 were most active against prostate and melanoma tumor cell lines, respectively. The compound did not present Topo I inhibitory activity. However, both derivatives displayed topoisomerase IIα inhibitory activity comparable to amsacrine, and AMTAC-02 was more potent than AMTAC-01 with methoxy substituent group on phenyl ring. This study demonstrates that the new derivatives are promising molecules with topoisomerase IIα inhibitory and antiproliferative activities.