Exploratory Data Analysis of Cell and Mitochondrial High-Fat, High-Sugar Toxicity on Human HepG2 Cells.

Non-alcoholic steatohepatitis (NASH), one of the deleterious stages of non-alcoholic fatty liver disease, remains a significant cause of liver-related morbidity and mortality worldwide. In the current work, we used an exploratory data analysis to investigate time-dependent cellular and mitochondrial effects of different supra-physiological fatty acids (FA) overload strategies, in the presence or absence of fructose (F), on human hepatoma-derived HepG2 cells. We measured intracellular neutral lipid content and reactive oxygen species (ROS) levels, mitochondrial respiration and morphology, and caspases activity and cell death. FA-treatments induced a time-dependent increase in neutral lipid content, which was paralleled by an increase in ROS. Fructose, by itself, did not increase intracellular lipid content nor aggravated the effects of palmitic acid (PA) or free fatty acids mixture (FFA), although it led to an up-expression of hepatic fructokinase. Instead, F decreased mitochondrial phospholipid content, as well as OXPHOS subunits levels. Increased lipid accumulation and ROS in FA-treatments preceded mitochondrial dysfunction, comprising altered mitochondrial membrane potential (ΔΨm) and morphology, and decreased oxygen consumption rates, especially with PA. Consequently, supra-physiological PA alone or combined with F prompted the activation of caspase pathways leading to a time-dependent decrease in cell viability. Exploratory data analysis methods support this conclusion by clearly identifying the effects of FA treatments. In fact, unsupervised learning algorithms created homogeneous and cohesive clusters, with a clear separation between PA and FFA treated samples to identify a minimal subset of critical mitochondrial markers in order to attain a feasible model to predict cell death in NAFLD or for high throughput screening of possible therapeutic agents, with particular focus in measuring mitochondrial function.

Cytotoxicity and Mitochondrial Effects of Phenolic and Quinone-Based Mitochondria-Targeted and Untargeted Antioxidants on Human Neuronal and Hepatic Cell Lines: A Comparative Analysis.

Mitochondriotropic antioxidants (MC3, MC6.2, MC4 and MC7.2) based on dietary antioxidants and analogs (caffeic, hydrocaffeic, trihydroxyphenylpropanoic and trihydroxycinnamic acids) were developed. In this study, we evaluate and compare the cytotoxicity profile of novel mitochondria-targeted molecules (generally known as MitoCINs) on human HepG2 and differentiated SH-SY5Y cells with the quinone-based mitochondria-targeted antioxidants MitoQ and SkQ1 and with two non-targeted antioxidants, resveratrol and coenzyme Q10 (CoQ10). We further evaluate their effects on mitochondrial membrane potential, cellular oxygen consumption and extracellular acidification rates. Overall, MitoCINs derivatives reduced cell viability at concentrations about six times higher than those observed with MitoQ and SkQ1. A toxicity ranking for both cell lines was produced: MC4 < MC7.2 < MC3 < MC6.2. These results suggest that C-6 carbon linker and the presence of a pyrogallol group result in lower cytotoxicity. MC3 and MC6.2 affected the mitochondrial function more significantly relative to MitoQ, SkQ1, resveratrol and CoQ10, while MC4 and MC7.2 displayed around 100-1000 times less cytotoxicity than SkQ1 and MitoQ. Based on the mitochondrial and cytotoxicity cellular data, MC4 and MC7.2 are proposed as leads that can be optimized to develop safe drug candidates with therapeutic application in mitochondrial oxidative stress-related diseases.

Single nanomolar doxorubicin exposure triggers compensatory mitochondrial responses in H9c2 cardiomyoblasts.

Dose-dependent and cumulative cardiotoxicity associated with doxorubicin (DOX) is the main limitation of anticancer therapy. Pediatric cancer survivors are particularly vulnerable, and no effective prevention measures are available. The aim of the present study was to investigate the persistent effects of nanomolar DOX concentrations and determine whether a pretreatment would induce mitochondrial adaptations in H9c2 cardiomyoblasts. H9c2 cells were incubated with DOX (10 and 25 nM) for 24 h, followed by 9 days of recovery in drug-free medium. We found that the sub-therapeutic DOX treatment induced persistent hypertrophy and dose-dependent cell cycle arrest in G2/M. Glycolytic activity, indirectly based on extracellular acidification rate, and basal respiration were significantly decreased in DOX-treated cells compared to controls, although both groups showed similar maximal respiration. Additionally, nanomolar DOX pretreatment resulted in upregulation of mitochondrial DNA transcripts accompanied by a decrease in DNA methyltransferase 1 (DNMT1) and global methylation levels. Finally, the pretreatment with DOX ameliorated H9c2 cells resistance against a subsequent exposure to DOX. These results suggest that nanomolar DOX pretreatment induced a beneficial and possibly epigenetic-based mitochondrial adaptation, raising the possibility that an early sub-therapeutic DOX treatment can be used as a preconditioning and protective approach during anticancer therapies.

A Mitochondrial Approach to Cardiovascular Risk and Disease.

BACKGROUND: Cardiovascular diseases (CVDs) are a leading risk factor for mortality worldwide and the number of CVDs victims is predicted to rise through 2030. While several external parameters (genetic, behavioral, environmental and physiological) contribute to cardiovascular morbidity and mortality; intrinsic metabolic and functional determinants such as insulin resistance, hyperglycemia, inflammation, high blood pressure and dyslipidemia are considered to be dominant factors. METHODS: Pubmed searches were performed using different keywords related with mitochondria and cardiovascular disease and risk. In vitro, animal and human results were extracted from the hits obtained. RESULTS: High cardiac energy demand is sustained by mitochondrial ATP production, and abnormal mitochondrial function has been associated with several lifestyle- and aging-related pathologies in the developed world such as diabetes, non-alcoholic fatty liver disease (NAFLD) and kidney diseases, that in turn can lead to cardiac injury. In order to delay cardiac mitochondrial dysfunction in the context of cardiovascular risk, regular physical activity has been shown to improve mitochondrial parameters and myocardial tolerance to ischemia-reperfusion (IR). Furthermore, pharmacological interventions can prevent the risk of CVDs. Therapeutic agents that can target mitochondria, decreasing ROS production and improve its function have been intensively researched. One example is the mitochondria-targeted antioxidant MitoQ10, which already showed beneficial effects in hypertensive rat models. Carvedilol or antidiabetic drugs also showed protective effects by preventing cardiac mitochondrial oxidative damage. CONCLUSION: This review highlights the role of mitochondrial dysfunction in CVDs, also show-casing several approaches that act by improving mitochondrial function in the heart, contributing to decrease some of the risk factors associated with CVDs.

Práticas de Enfermagem na Coordenação do Cuidado na Atenção Primária à Saúde / Nursing Practices In Coordination Care In Primary Health Care / Prácticas de Enfermería Coordinación Del Cuidado En La Atención Primaria da Salud

Objetivo: analisar as características das equipes e as práticas associadas ao acompanhamento e coordenação do cuidado no Pará e compreender como ocorre essa prática executada pelo enfermeiro como membro da equipe na Atenção Primária à Saúde. Métodos: estudo de métodos mistos. Os participantes foram enfermeiros da atenção básica do Pará. A coleta ocorreu entre novembro de 2019 a agosto de 2021, através de um formulário eletrônico e entrevista. A análise integrativa dos dados foi feita pela conexão dos métodos qualitativos e quantitativos. Resultados: A proporção de enfermeiros que acompanha e coordena os usuários que estão em uso de outros serviços foi de 50% (90/180). Houve associação significativa (p <0,05) entre o acompanhamento e coordenação do cuidado e o tipo de equipe, carga horária de trabalho, regulação das demandas locais na perspectiva da rede, articulação com profissionais de saúde de outros níveis de atenção, trabalhos com profissionais de outras formações e a condução da clínica ampliada/ matriciamento. As categorias temáticas "relação da Atenção Primária à Saúde com a rede de atenção" e "práticas de coordenação e continuidade do cuidado horizontal e vertical" apresentaram convergência com os dados quantitativos e a categoria temática "Práticas de coordenação do cuidado sob a forma do trabalho em equipe" converge com os dados quantitativos, mas divergem no apoio matricial. Conclusão: Os enfermeiros realizam o acompanhamento e coordenação do cuidado, porém enfrentam dificuldades que resultam em sobrecarga e realização de atribuições que não são da categoria profissional. (AU)

Objective: to analyze the characteristics of the teams and the practices associated with the monitoring and coordination of care in Pará and to understand how this practice is carried out by the nurse as a member of the team in Primary Health Care. Methods: study of mixed methods. The participants were primary care nurses in Pará. The collection took place between November 2019 and August 2021, through an electronic form and interview. The integrative data analysis was performed by connecting qualitative and quantitative methods. Results: The proportion of nurses who monitor and coordinate users who are using other services was 50% (90/180). There was a significant association (p <0.05) between monitoring and coordination of care and the type of team, workload, regulation of local demands from the perspective of the network, articulation with health professionals from other levels of care, work with professionals from other backgrounds and conducting the expanded clinic/matrix support. The thematic categories "relationship between Primary Health Care and the care network" and "practices of coordination and continuity of horizontal and vertical care" showed convergence with the quantitative data and the thematic category "Practices of coordination of care in the form of work in a team" converges with the quantitative data, but differs in matrix support. Conclusion: Nurses carry out monitoring and coordination of care, but they face difficulties that result in overload and carrying out tasks that are not of the professional category. (AU)

Objetivo: analizar las características de los equipos y las prácticas asociadas al seguimiento y coordinación del cuidado en Pará y comprender cómo esa práctica es realizada por el enfermero como miembro del equipo en la Atención Primaria de Salud. Métodos: estudio de métodos mixtos. Los participantes eran enfermeros de atención primaria de Pará. La recolección se realizó entre noviembre de 2019 y agosto de 2021, a través de formulario electrónico y entrevista. El análisis integrador de datos se realizó conectando métodos cualitativos y cuantitativos. Resultados: La proporción de enfermeros que acompañan y coordinan usuarios que utilizan otros servicios fue del 50% (90/180). Hubo asociación significativa (p<0,05) entre el seguimiento y coordinación de la atención y el tipo de equipo, carga de trabajo, regulación de las demandas locales desde la perspectiva de la red, articulación con profesionales de salud de otros niveles de atención, trabajo con profesionales de otros antecedentes y dirigiendo el apoyo clínico/matriz ampliado. Las categorías temáticas "relación entre la Atención Primaria de Salud y la red de atención" y "prácticas de coordinación y continuidad del cuidado horizontal y vertical" mostraron convergencia con los datos cuantitativos y la categoría temática "Prácticas de coordinación del cuidado en la forma de trabajo en un team" converge con los datos cuantitativos, pero difiere en el soporte de la matriz. Conclusión: Los enfermeros realizan seguimiento y coordinación de los cuidados, pero enfrentan dificultades que resultan en sobrecarga y realización de tareas que no son de categoría profesional. (AU)

Práticas coletivas e individuais associadas à dificuldade dos enfermeiros da atenção primária à saúde / Collective and individual practices associated with the difficulty of primary health care nurses / Prácticas coletivas e individuais asociadas a la dificultad de las enfermeras de atención primaria de salud

Objetivo: analisar as práticas individuais e coletivas que estão associadas as dificuldades dos enfermeiros que atuam na Atenção Primária à Saúde da região Norte. Métodos: observacional de prevalência e analítico. Foram incluídos enfermeiros da atenção básica. A coleta ocorreu entre novembro de 2019 a agosto de 2021, nos sete estados do Norte, através de um formulário eletrônico, analisados pelos testes binomial, quiquadrado e G no programa Bioestat. Resultados: Entre os 626 enfermeiros do estudo, 15,7% (98/626) afirmaram ter dificuldade no exercício de suas práticas, quanto à autonomia das suas responsabilidades normativas legais. O Amapá teve menor proporção (8,3%; 9/108) de enfermeiros com dificuldade em relação a região. As práticas associadas a dificuldade foram: a não participação do enfermeiro no gerenciamento dos insumos (p=0,03), realizar consulta eventualmente (p=0,03) e nunca prescrever medicamentos (p=0,02); resolutividade insuficiente na consulta pré-natal (p=0,000), acompanhamento de crescimento e desenvolvimento infantil (p=0,001); planejamento familiar (p=0,000); hanseníase (p=0,005); tuberculose (p=0,031); hipertensão arterial (p<0,0001); diabetes (p<0,0001). Conclusão: As dificuldades quanto à autonomia das responsabilidades normativas legais estão associadas a práticas individuais e coletivas que são privativas do enfermeiro e estão regulamentadas nos programas de saúde pública. (AU)

Objective: to analyze the individual and collective practices that are associated with the difficulties of nurses who work in Primary Health Care in the North region. Methods: observational of prevalence and analytical. Primary care nurses were included. The collection took place between November 2019 and August 2021, in the seven states of the North, through an electronic form, analyzed by the binomial, chi-square and G tests in the Bioestat program. Results: Among the 626 nurses in the study, 15.7% (98/626) stated that they had difficulty in exercising their practices, regarding the autonomy of their legal normative responsibilities. Amapá had a lower proportion (8.3%; 9/108) of nurses with difficulties in relation to the region. The practices associated with the difficulty were: the non-participation of nurses in the management of supplies (p=0.03), occasional consultations (p=0.03) and never prescribing medication (p=0.02); insufficient resolution in the prenatal consultation (p=0.000), monitoring of child growth and development (p=0.001); family planning (p=0.000); leprosy (p=0.005); tuberculosis (p=0.031); arterial hypertension (p<0.0001); diabetes (p<0.0001). Conclusion: Difficulties regarding the autonomy of legal normative responsibilities are associated with individual and collective practices that are exclusive to nurses and are regulated in public health programs. (AU)

Objetivo: analizar las prácticas individuales y colectivas que están asociadas a las dificultades de los enfermeros que actúan en la Atención Primaria de Salud de la región Norte. Métodos: observacional de prevalencia y analítico. Se incluyeron enfermeras de atención primaria. La colecta se realizó entre noviembre de 2019 y agosto de 2021, en los siete estados del Norte, a través de un formulario electrónico, analizado por las pruebas binomial, chi-cuadrado y G en el programa Bioestat. Resultados: Entre los 626 enfermeros del estudio, 15,7% (98/626) afirmaron tener dificultad en el ejercicio de sus prácticas, en cuanto a la autonomía de sus responsabilidades normativas legales. Amapá tuvo menor proporción (8,3%; 9/108) de enfermeros con dificultades en relación a la región. Las prácticas asociadas a la dificultad fueron: la no participación de los enfermeros en la gestión de los insumos (p=0,03), consultas ocasionales (p=0,03) y nunca recetar medicamentos (p=0,02); resolución insuficiente en la consulta prenatal (p=0,000), seguimiento del crecimiento y desarrollo infantil (p=0,001); planificación familiar (p=0,000); lepra (p=0,005); tuberculosis (p=0,031); hipertensión arterial (p<0,0001); diabetes (p<0,0001). Conclusión: Las dificultades en cuanto a la autonomía de las responsabilidades normativas legales están asociadas a las prácticas individuales y colectivas, exclusivas de los enfermeros y reguladas em los programas de salud pública. (AU)

Autonomia do enfermeiro no exercício das práticas de enfermagem na atenção primária à saúde / Nurses autonomy in the exercise of nursing practices in primary health care / Autonomía de las enfermeras en el ejercicio de las prácticas de enfermería en la atención primaria de salud

Objetivo: Compreender a autonomia do enfermeiro quanto as suas responsabilidades normativas legais no exercício das práticas de enfermagem na Atenção Primária à Saúde (APS) do Pará. Métodos: Estudo observacional, de delineamento transversal qualitativo, realizado em três municípios do estado do Pará. A coleta de dados foi realizada de outubro de 2020 a janeiro de 2021. Os participantes foram os enfermeiros da Atenção Primária à Saúde. Foi realizada entrevista, com roteiro semiestruturado. Foi realizada análise temática dos conteúdos, com auxílio da ferramenta IRaMuTeq Resultados: Emergiram cinco categorias temáticas: 1) Autonomia do enfermeiro no contexto da prescrição de outro profissional na APS; 2) Autonomia para prescrição nos programas e exames na APS; 3) Áreas de identificação da autonomia profissional na APS; 4) As práticas preventivas na APS e suas dificuldades; e 5) Limites da prática profissional do enfermeiro na APS. Conclusão: Os enfermeiros da APS do Pará têm práticas de enfermagem individuais com diferentes tipos de autonomia, cujos cuidados colocam em prática com o respaldo dos protocolos e regulamentos técnicos. Contudo, há necessidade de ampliar e fortalecer parcerias com outros atores sociais municipais. (AU)

Objective: To understand the autonomy of nurses regarding their legal regulatory responsibilities in the exercise of nursing practices in Primary Health Care (PHC) in Pará. Methods: Observational study with qualitative cross-sectional design, conducted in three municipalities of the state of Pará. Data collection was carried out from October 2020 to January 2021. The participants were Primary Health Care nurses. Interviews were conducted, with a semi-structured script. A thematic analysis of the contents was performed, with the help of the IRaMuTeq tool. Results: Five thematic categories emerged: 1) Nurses' autonomy in the context of another professional's prescription in PHC; 2) Autonomy for prescription in programs and exams in PHC; 3) Areas of identification of professional autonomy in PHC; 4) Preventive practices in PHC and their difficulties; and 5) Limits of nurses' professional practice in PHC. Conclusion: PHC nurses in Pará have individual nursing practices with different types of autonomy, whose care they put into practice with the support of protocols and technical regulations. However, there is a need to expand and strengthen partnerships with other municipal social actors. (AU)

Objetivo: Comprender la autonomía de los enfermeros en cuanto a sus responsabilidades normativas legales en el ejercicio de las prácticas de enfermería en la Atención Básica a la Salud (APS) en Pará. Métodos: Estudio observacional, con diseño transversal cualitativo, realizado en tres municipios del estado de Pará. La recolección de datos se realizó de octubre de 2020 a enero de 2021. Los participantes fueron enfermeros de la Atención Primaria de Salud. Se realizó una entrevista, con un guión semiestructurado. Se realizó un análisis temático de dos contenidos, con la ayuda de la herramienta IRaMuTeq. Resultados: Emergieron cinco categorías temáticas: 1) Autonomía del enfermero en el contexto de la prescripción de otro profesional en la APS; 2) Autonomía para prescribir programas y exámenes en la APS; 3) Áreas de identificación de la autonomía profesional en APS; 4) Prácticas preventivas en APS y sus dificultades; y 5) Límites del ejercicio profesional de enfermería en la APS. Conclusión: Los enfermeros de la APS de Pará tienen prácticas de enfermería individuales con diferentes tipos de autonomía, cuyo cuidado está sustentado por dos protocolos y normas técnicas. Sin embargo, existe la necesidad de ampliar y fortalecer las alianzas con otros socios municipales. (AU)