Dietary interventions in mice affect oxidative stress and gene expression of the Prlr and Esr1 in the adipose tissue and hypothalamus of dams and their offspring

Maternal diet is key to the progeny’s health since it may impact on the offspring’s adult life. In this study, mice dams received standard (CONT), restrictive (RD), or hypercaloric (HD) diets during mating, pregnancy, and lactation. Male offspring of each group of dams also received these diets: CONT, RD, HD. Aiming to evaluate the oxidative stress in the adipose tissue, reactive oxygen species (ROS) production, catalase (CAT), and superoxide dismutase (SOD) activities were analyzed in dams and offspring. In the adipose tissue and hypothalamus, gene expression of prolactin (Prlr) and estrogen alpha (Esr1) receptors was performed in dams and offspring. Protein expression of Stat5 was evaluated in the adipose tissue of the offspring from RD-fed dams. HD-fed dams increased triglycerides and leptin serum concentrations, and decreased SOD activity in the adipose tissue. In the offspring’s adipose tissue, we observed a maternal diet effect caused by HD, with increased ROS production and SOD and CAT activities. Gene expression of Prlr and Esr1 in the offspring’s adipose tissue was decreased due to maternal RD. Mice from HD-fed dams showed higher Stat5 expression compared to the offspring from CONT and RD dams in the adipose tissue. In the hypothalamus, we found decreased expression of Prlr in RD and HD dams, compared to CONT; and a maternal diet effect on Prlr and Esr1 gene expression in the offspring. In conclusion, we can affirm that maternal nutrition impacts the redox state and influences the gene expression of Prlr and Esr1, which are involved in energy metabolism, both peripherally and centrally in the adult life of the female offspring. (AU)

Gene expression evaluation of antioxidant enzymes in patients with hepatocellular carcinoma: RT-qPCR and bioinformatic analyses.

Any condition leading to chronic liver disease is a potential oncogenic agent for hepatocellular carcinoma (HCC). Alterations in the expression of antioxidant enzymes could alter the redox balance. Our aim was to evaluate the expression of the genes GPX1, GPX4, SEP15, SELENOP, SOD1, SOD2, GSR, CAT, and NFE2L2 in patients with HCC. Differential gene expression analysis was performed using RNA-Seq data from the TCGA and GTEx databases, and RT-qPCR data from HCC patient samples. Bioinformatic analysis revealed significant differential expression in most genes. GPX4 expression was significantly increased (p=0.02), while SOD2 expression was significantly decreased (p=0.04) in experimental data. In TCGA samples, alpha-fetoprotein levels (mg/dL) were negatively correlated with the expression of SEP15 (p<0.001), SELENOP (p<0.001), SOD1 (p<0.001), SOD2 (p<0.001), CAT (p<0.001), and NFE2L2 (p=0.004). Alpha-fetoprotein levels were positively correlated with the expression of GPX4 (p=0.02) and SELENOP (p=0.01) in the experimental data. Low expression of GPX1 (p=0.006), GPX4 (p=0.01), SELENOP (p=0.006), SOD1 (p=0.007), CAT (p<0.001), and NFE2L2 (p<0.001), and higher levels of GSR, were associated with low overall survival at 12 months. These results suggest a significant role for these antioxidant enzymes in HCC pathogenesis and severity.

Transcriptional expression study in the central nervous system of rats: what gene should be used as internal control?

OBJECTIVE: A growing number of published articles report the expression of specific genes with different behavior patterns in rats. The levels of messenger ribonucleic acid transcripts are usually analyzed by reverse transcription followed by polymerase chain reaction and quantified after normalization with an internal control or reference gene (housekeeping gene). Nevertheless, housekeeping genes exhibit different expression in the central nervous system, depending on the physiological conditions and the area of the brain to be studied. The choice of a good internal control gene is essential for obtaining reliable results. This study evaluated the expression of three housekeeping genes (beta-actin, cyclophilin A, and ubiquitin C) in different areas of the central nervous system in rats (olfactory bulb, hippocampus, striatum, and prefrontal cortex). METHODS: Wistar rats (virgin females, n=6) during the diestrum period were used. Total ribonucleic acid was extracted from each region of the brain; the complementary deoxyribonucleic acid was synthesized by reverse transcription and amplified by real-time quantitative polymerase chain reaction using SYBR™ Green and primers specific for each one of the reference genes. The stability of the expression was determined using NormFinder. RESULTS: Beta-actin was the most stable gene in the hippocampus and striatum, while cyclophilin A and ubiquitin C showed greater stability in the prefrontal cortex and the olfactory bulb, respectively. CONCLUSION: Based on our study, further studies of gene expression using rats as animal models should take into consideration these results when choosing a reliable internal control gene.

Transcriptional expression study in the central nervous system of rats: what gene should be used as internal control? / Estudo de expressão transcricional no sistema nervoso central de ratos: qual gene deve ser usado como controle interno?

Objective A growing number of published articles report the expression of specific genes with different behavior patterns in rats. The levels of messenger ribonucleic acid transcripts are usually analyzed by reverse transcription followed by polymerase chain reaction and quantified after normalization with an internal control or reference gene (housekeeping gene). Nevertheless, housekeeping genes exhibit different expression in the central nervous system, depending on the physiological conditions and the area of the brain to be studied. The choice of a good internal control gene is essential for obtaining reliable results. This study evaluated the expression of three housekeeping genes (beta-actin, cyclophilin A, and ubiquitin C) in different areas of the central nervous system in rats (olfactory bulb, hippocampus, striatum, and prefrontal cortex). Methods Wistar rats (virgin females, n=6) during the diestrum period were used. Total ribonucleic acid was extracted from each region of the brain; the complementary deoxyribonucleic acid was synthesized by reverse transcription and amplified by real-time quantitative polymerase chain reaction using SYBR™ Green and primers specific for each one of the reference genes. The stability of the expression was determined using NormFinder. Results Beta-actin was the most stable gene in the hippocampus and striatum, while cyclophilin A and ubiquitin C showed greater stability in the prefrontal cortex and the olfactory bulb, respectively. Conclusion Based on our study, further studies of gene expression using rats as animal models should take into consideration these results when choosing a reliable internal control gene. .

Objetivo Um número crescente de artigos publicados relaciona a expressão de genes específicos com diferentes padrões de comportamento em ratos. Os níveis de transcritos de ácido ribonucleico mensageiro são geralmente analisados por transcrição reversa, seguida de reação em cadeia da polimerase, e quantificados após a normalização com um controle interno ou gene de referência (gene housekeeping). No entanto, os genes housekeeping exibem expressão diferencial no sistema nervoso central, dependendo das condições fisiológicas e da área do cérebro a ser estudada. A escolha de um bom gene de controle interno é essencial para a obtenção de resultados confiáveis. Este estudo avaliou a expressão de três genes housekeeping (beta-actina, ciclofilina A e ubiquitina C) em diferentes áreas do sistema nervoso central de ratos (bulbo olfatório, hipocampo, estriado e córtex pré-frontal). Métodos Foram usadas ratas Wistar (fêmeas virgens, n=6) durante o período de diestro. O ácido ribonucleico total foi extraído a partir de cada região do cérebro; o ácido desoxirribonucleico complementar foi sintetizado por transcrição reversa e amplificado por reação em cadeia da polimerase quantitativo em tempo real utilizando SYBR® Green e primers específicos para cada um dos genes de referência. A estabilidade de expressão foi determinada utilizando NormFinder. Resultados A beta-actina foi o gene mais estável no hipocampo e estriado, enquanto a ciclofilina A e a ubiquitina C apresentaram maior estabilidade no córtex pré-frontal e no bulbo olfatório, respectivamente. Conclusão Com base em nosso trabalho, estudos posteriores de expressão gênica utilizando ratos como modelos animais devem levar ...

Nós reflexivos: a cartografia como estratégia metodológica / Reflective knots: cartography as a methodological strategy / Nudos reflexivos: la cartografía como estrategia metodologica

Este artigo traz aproximações do método cartográfico a partir dos estudos de Virgínia Kastrup, com o conceito de enação e de observador implicado, proposições de Francisco Varela e Humberto Maturana, autores que orientam os caminhos investigativos produzidos. Que movimento é esse que faz a pesquisa? Como acompanhar processos? Como compreender que os processos nos transformam? O que significa dizer que os processos produzem mundos? Com base nesses questionamentos o artigo compartilha um processo em vez de trazer resultados, cartografa o caminhar na pesquisa e produz nós reflexivos, pontos de encontro e convergência das experiências, emoções e saberes das pesquisadoras. Tais nós reflexivos são conceitos e compreensões que interferem e rompem a trajetória da pesquisa e provocam a reconstrução dos caminhos mesmo antes deles existirem. (AU)

This paper presents approaches approximations of cartographic method from Virginia Kastrup’s studies, with the concept of enaction and implicated observer, propositions of Francisco Varela and Humberto Maturana, authors who guided the investigative paths produced. What is this movement that makes research? How to monitor processes? How to understand that processes change us? What does it means stating processes change worlds? Based on these questions the article shares a process instead of bringing results, maps the research walk and produces reflective knots, meeting points and convergence of experiences, emotions and knowledge of the researchers. Such reflective knots are concepts and understandings that interfere and disrupt the course of the research and cause the reconstruction of roads even before they exist. (AU)

Este artículo trae aproximaciones del método cartográfico a partir de los estúdios de Virgínia Kastrup, com el concepto de en acción y de observador implicado, proposiciones de Francisco Varela y Humberto Maturana, autores que orientan los caminos investigativos producidos. Qué movimiento es ese que hace la investigación? Como acompañar procesos? Como comprender que los procesos nos transforman? Que significa decir que los procesos producen mundos? Con base en esos cuestionamientos el artículo comparte un proceso en vez de traer resultados, cartografía el caminar en la investigación y produce nudos reflexivos, puntos de encuentro y convergencia de las experiencias, emociones y saberes de las investigadoras. Tales nudos reflexivos son conceptos y comprensiones que interfieren y rompen la trayectoria de la investigación y provocan la reconstrucción de los caminos incluso antes de que existan. (AU)

Nós reflexivos: a cartografia como estratégia metodológica / Reflective knots: cartography as a methodological strategy / Nudos reflexivos: la cartografía como estrategia metodologica

Este artigo traz aproximações do método cartográfico a partir dos estudos de Virgínia Kastrup, com o conceito de enação e de observador implicado, proposições de Francisco Varela e Humberto Maturana, autores que orientam os caminhos investigativos produzidos. Que movimento é esse que faz a pesquisa? Como acompanhar processos? Como compreender que os processos nos transformam? O que significa dizer que os processos produzem mundos? Com base nesses questionamentos o artigo compartilha um processo em vez de trazer resultados, cartografa o caminhar na pesquisa e produz nós reflexivos, pontos de encontro e convergência das experiências, emoções e saberes das pesquisadoras. Tais nós reflexivos são conceitos e compreensões que interferem e rompem a trajetória da pesquisa e provocam a reconstrução dos caminhos mesmo antes deles existirem.

This paper presents approaches approximations of cartographic method from Virginia Kastrup’s studies, with the concept of enaction and implicated observer, propositions of Francisco Varela and Humberto Maturana, authors who guided the investigative paths produced. What is this movement that makes research? How to monitor processes? How to understand that processes change us? What does it means stating processes change worlds? Based on these questions the article shares a process instead of bringing results, maps the research walk and produces reflective knots, meeting points and convergence of experiences, emotions and knowledge of the researchers. Such reflective knots are concepts and understandings that interfere and disrupt the course of the research and cause the reconstruction of roads even before they exist.

Este artículo trae aproximaciones del método cartográfico a partir de los estúdios de Virgínia Kastrup, com el concepto de en acción y de observador implicado, proposiciones de Francisco Varela y Humberto Maturana, autores que orientan los caminos investigativos producidos. Qué movimiento es ese que hace la investigación? Como acompañar procesos? Como comprender que los procesos nos transforman? Que significa decir que los procesos producen mundos? Con base en esos cuestionamientos el artículo comparte un proceso en vez de traer resultados, cartografía el caminar en la investigación y produce nudos reflexivos, puntos de encuentro y convergencia de las experiencias, emociones y saberes de las investigadoras. Tales nudos reflexivos son conceptos y comprensiones que interfieren y rompen la trayectoria de la investigación y provocan la reconstrucción de los caminos incluso antes de que existan.

(Re)construindo a experiência de mulheres com HIV/AIDS no grupo de apoio do CRASE, em Governador Valadares, MG / (Re)building the life history for adhesion to the treatment: the experience of women with HIV/AIDS

Trata-se de estudo etnográfico cujo objetivo foi compreender a experiência da adesão ao tratamento por mulheres com HIV/AIDS...