Exposure to toxicologically relevant atrazine concentrations impair the glycolytic function of mouse Sertoli cells through the downregulation of lactate dehydrogenase.

Atrazine (ATZ), a widely used herbicide with potent endocrine-disrupting properties, has been implicated in hormonal disturbances and fertility issues. Sertoli cells (SCs) play a crucial role in providing mechanical and nutritional support of spermatogenesis. Herein, we aimed to study the effects of environmentally relevant ATZ concentrations on the nutritional support of spermatogenesis provided by SCs. For that, mouse SCs (TM4) were exposed to increasing ATZ concentrations (in µg/L: 0.3, 3, 30, 300, or 3000). After 24 h, cellular proliferation and metabolic activity were assessed. Mitochondrial activity and endogenous reactive oxygen species (ROS) production were evaluated using JC-1 and CM-H2DCFDA probes, respectively. We also analyzed protein levels of lactate dehydrogenase (LDH) using Western Blot and live cells glycolytic function through Seahorse XF Glycolysis Stress Test Kit. ATZ exposure decreased the activity of oxidoreductases in SCs, suggesting a decreased metabolic activity. Although ATZ is reported to induce oxidative stress, we did not observe alterations in mitochondrial membrane potential and ROS production across all tested concentrations. When we evaluated the glycolytic function of SCs, we observed that ATZ significantly impaired glycolysis and the glycolytic capacity at all tested concentrations. These results were supported by the decreased expression of LDH in SCs. Overall, our findings suggest that ATZ impairs the glycolytic function of SCs through LDH downregulation. Since lactate is the preferential energetic substrate for germ cells, exposure to ATZ may detrimentally impact the nutritional support crucial for spermatogenesis, hinting for a relationship between ATZ exposure and male infertility.

Weaknesses in the continuity of care for preterm infants discharged from the neonatal unit.

OBJECTIVE: To identify weaknesses in the continuity of care for preterm infants discharged from a neonatal unit, based on the perspective of professionals in the family health strategy. METHOD: Qualitative research, carried out with 16 professionals from four health regions in a capital city in the center-west of Brazil. Data collection took place from October to December 2020, through semi-structured, individual, and in-person interviews. Data underwent content analysis, supported by the concept of continuity of care. RESULTS: The analysis consisted of three categories: Challenges for care in the unit and referral to specialized services; weak interactions between the preterm baby's family and health professionals; Information: essential aspect for the connection between health professionals and the family of the preterm newborn. CONCLUSION: Health services are shown to be fragile in terms of the dimensions of continuity of care, contributing to the discontinuity of care for preterm children.

Zika virus infection impairs synaptogenesis, induces neuroinflammation, and could be an environmental risk factor for autism spectrum disorder outcome.

Zika virus (ZIKV) infection was first associated with Central Nervous System (CNS) infections in Brazil in 2015, correlated with an increased number of newborns with microcephaly, which ended up characterizing the Congenital Zika Syndrome (CZS). Here, we investigated the impact of ZIKV infection on the functionality of iPSC-derived astrocytes. Besides, we extrapolated our findings to a Brazilian cohort of 136 CZS children and validated our results using a mouse model. Interestingly, ZIKV infection in neuroprogenitor cells compromises cell migration and causes apoptosis but does not interfere in astrocyte generation. Moreover, infected astrocytes lost their ability to uptake glutamate while expressing more glutamate transporters and secreted higher levels of IL-6. Besides, infected astrocytes secreted factors that impaired neuronal synaptogenesis. Since these biological endophenotypes were already related to Autism Spectrum Disorder (ASD), we extrapolated these results to a cohort of children, now 6-7 years old, and found seven children with ASD diagnosis (5.14 %). Additionally, mice infected by ZIKV revealed autistic-like behaviors, with a significant increase of IL-6 mRNA levels in the brain. Considering these evidence, we inferred that ZIKV infection during pregnancy might lead to synaptogenesis impairment and neuroinflammation, which could increase the risk for ASD.

Which terms should be used to describe medications used in the treatment of seizure disorders? An ILAE position paper.

A variety of terms, such as "antiepileptic," "anticonvulsant," and "antiseizure" have been historically applied to medications for the treatment of seizure disorders. Terminology is important because using terms that do not accurately reflect the action of specific treatments may result in a misunderstanding of their effects and inappropriate use. The present International League Against Epilepsy (ILAE) position paper used a Delphi approach to develop recommendations on English-language terminology applicable to pharmacological agents currently approved for treating seizure disorders. There was consensus that these medications should be collectively named "antiseizure medications". This term accurately reflects their primarily symptomatic effect against seizures and reduces the possibility of health care practitioners, patients, or caregivers having undue expectations or an incorrect understanding of the real action of these medications. The term "antiseizure" to describe these agents does not exclude the possibility of beneficial effects on the course of the disease and comorbidities that result from the downstream effects of seizures, whenever these beneficial effects can be explained solely by the suppression of seizure activity. It is acknowledged that other treatments, mostly under development, can exert direct favorable actions on the underlying disease or its progression, by having "antiepileptogenic" or "disease-modifying" effects. A more-refined terminology to describe precisely these actions needs to be developed.

Live, Attenuated, Tetravalent Butantan-Dengue Vaccine in Children and Adults.

BACKGROUND: Butantan-Dengue Vaccine (Butantan-DV) is an investigational, single-dose, live, attenuated, tetravalent vaccine against dengue disease, but data on its overall efficacy are needed. METHODS: In an ongoing phase 3, double-blind trial in Brazil, we randomly assigned participants to receive Butantan-DV or placebo, with stratification according to age (2 to 6 years, 7 to 17 years, and 18 to 59 years); 5 years of follow-up is planned. The objectives of the trial were to evaluate overall vaccine efficacy against symptomatic, virologically confirmed dengue of any serotype occurring more than 28 days after vaccination (the primary efficacy end point), regardless of serostatus at baseline, and to describe safety up to day 21 (the primary safety end point). Here, vaccine efficacy was assessed on the basis of 2 years of follow-up for each participant, and safety as solicited vaccine-related adverse events reported up to day 21 after injection. Key secondary objectives were to assess vaccine efficacy among participants according to dengue serostatus at baseline and according to the dengue viral serotype; efficacy according to age was also assessed. RESULTS: Over a 3-year enrollment period, 16,235 participants received either Butantan-DV (10,259 participants) or placebo (5976 participants). The overall 2-year vaccine efficacy was 79.6% (95% confidence interval [CI], 70.0 to 86.3) - 73.6% (95% CI, 57.6 to 83.7) among participants with no evidence of previous dengue exposure and 89.2% (95% CI, 77.6 to 95.6) among those with a history of exposure. Vaccine efficacy was 80.1% (95% CI, 66.0 to 88.4) among participants 2 to 6 years of age, 77.8% (95% CI, 55.6 to 89.6) among those 7 to 17 years of age, and 90.0% (95% CI, 68.2 to 97.5) among those 18 to 59 years of age. Efficacy against DENV-1 was 89.5% (95% CI, 78.7 to 95.0) and against DENV-2 was 69.6% (95% CI, 50.8 to 81.5). DENV-3 and DENV-4 were not detected during the follow-up period. Solicited systemic vaccine- or placebo-related adverse events within 21 days after injection were more common with Butantan-DV than with placebo (58.3% of participants, vs. 45.6%). CONCLUSIONS: A single dose of Butantan-DV prevented symptomatic DENV-1 and DENV-2, regardless of dengue serostatus at baseline, through 2 years of follow-up. (Funded by Instituto Butantan and others; DEN-03-IB ClinicalTrials.gov number, NCT02406729, and WHO ICTRP number, U1111-1168-8679.).

Live, attenuated, tetravalent Butantan- Dengue vaccine in children and adults

BACKGROUND Butantan–Dengue Vaccine (Butantan-DV) is an investigational, single-dose, live, attenuated, tetravalent vaccine against dengue disease, but data on its overall efficacy are needed. METHODS In an ongoing phase 3, double-blind trial in Brazil, we randomly assigned participants to receive Butantan-DV or placebo, with stratification according to age (2 to 6 years, 7 to 17 years, and 18 to 59 years); 5 years of follow-up is planned. The objectives of the trial were to evaluate overall vaccine efficacy against symptomatic, virologically con firmed dengue of any serotype occurring more than 28 days after vaccination (the primary efficacy end point), regardless of serostatus at baseline, and to describe safety up to day 21 (the primary safety end point). Here, vaccine efficacy was assessed on the basis of 2 years of follow-up for each participant, and safety as solicited vaccine-related adverse events reported up to day 21 after injection. Key secondary objectives were to assess vaccine efficacy among participants according to dengue serostatus at baseline and according to the dengue viral serotype; efficacy according to age was also assessed. RESULTS Over a 3-year enrollment period, 16,235 participants received either Butantan-DV (10,259 participants) or placebo (5976 participants). The overall 2-year vaccine effi cacy was 79.6% (95% confidence interval [CI], 70.0 to 86.3) — 73.6% (95% CI, 57.6 to 83.7) among participants with no evidence of previous dengue exposure and 89.2% (95% CI, 77.6 to 95.6) among those with a history of exposure. Vaccine effi cacy was 80.1% (95% CI, 66.0 to 88.4) among participants 2 to 6 years of age, 77.8% (95% CI, 55.6 to 89.6) among those 7 to 17 years of age, and 90.0% (95% CI, 68.2 to 97.5) among those 18 to 59 years of age. Efficacy against DENV-1 was 89.5% (95% CI, 78.7 to 95.0) and against DENV-2 was 69.6% (95% CI, 50.8 to 81.5). DENV-3 and DENV-4 were not detected during the follow-up period. Solicited systemic vaccine- or placebo-related adverse events within 21 days after injection were more common with Butantan-DV than with placebo (58.3% of participants, vs. 45.6%). CONCLUSIONS A single dose of Butantan-DV prevented symptomatic DENV-1 and DENV-2, regard less of dengue serostatus at baseline, through 2 years of follow-up. (Funded by Instituto Butantan and others; DEN-03-IB ClinicalTrials.gov number, NCT02406729, and WHO ICTRP number, U1111-1168-8679.)

Weaknesses in the continuity of care for preterm infants discharged from the neonatal unit / Debilidades en la continuidad de la atención a los recién nacidos prematuros egresados de la unidad neonatal / Fragilidades para a continuidade do cuidado ao pré-termo egressoda unidade neonatal

ABSTRACT Objective: To identify weaknesses in the continuity of care for preterm infants discharged from a neonatal unit, based on the perspective of professionals in the family health strategy. Method: Qualitative research, carried out with 16 professionals from four health regions in a capital city in the center-west of Brazil. Data collection took place from October to December 2020, through semi-structured, individual, and in-person interviews. Data underwent content analysis, supported by the concept of continuity of care. Results: The analysis consisted of three categories: Challenges for care in the unit and referral to specialized services; weak interactions between the preterm baby's family and health professionals; Information: essential aspect for the connection between health professionals and the family of the preterm newborn. Conclusion: Health services are shown to be fragile in terms of the dimensions of continuity of care, contributing to the discontinuity of care for preterm children.

RESUMEN Objetivo: Identificar debilidades en la continuidad de la atención al recién nacido prematuro egresado de una unidad neonatal, desde la perspectiva de los profesionales de la estrategia de salud de la familia. Método: Investigación cualitativa, realizada con 16 profesionales de cuatro regiones sanitarias de una capital del centro-oeste de Brasil. La recolección de datos se realizó de octubre a diciembre de 2020, mediante entrevistas semiestructuradas, individuales y presenciales. Los datos fueron sometidos a análisis de contenido, sustentado en el concepto de continuidad de la atención. Resultados: Tres categorías comprendieron el análisis: Desafíos para la atención en la unidad y derivación a servicios especializados; Interacciones débiles entre la familia del bebé prematuro y los profesionales de la salud; Información: aspecto esencial para la vinculación entre los profesionales de la salud y la familia del recién nacido prematuro. Conclusión: Los servicios de salud son frágiles en términos de continuidad de la atención, lo que contribuye a la discontinuidad de la atención a los niños nacidos prematuros.

RESUMO Objetivo: Identificar as fragilidades para a continuidade do cuidado ao pré-termo egresso de unidade neonatal, a partir da perspectiva de profissionais da estratégia saúde da família. Método: Pesquisa qualitativa, realizada junto a 16 profissionais de quatro regionais de saúde de uma capital do centro-oeste do Brasil. A coleta dos dados ocorreu nos meses de outubro a dezembro de 2020, por meio de entrevistas semiestruturadas, individuais e presenciais. Os dados foram submetidos à analise de conteúdo, sustentada pelo conceito da continuidade do cuidado. Resultados: Três categorias compuseram a análise: Desafios para o atendimento na unidade e para o encaminhamento aos serviços especializados; Interações frágeis entre família do pré-termo e profissionais de saúde; Informação: aspecto essencial para a conexão entre profissionais de saúde e família do recém-nascido pré-termo. Conclusão: Os serviços de saúde mostram-se frágeis quanto às dimensões da continuidade do cuidado colaborando para a descontinuidade da atenção à criança nascida pré-termo.

Coping familiar no cuidado à criança com síndrome de Down: revisão de escopo / Family coping in caring for children with Down syndrome: a scoping review

RESUMO Objetivo mapear as estratégias de coping que contribuem para o cuidado familiar à criança com síndrome de Down. Métodos revisão de escopo, segundo o método JBI. Foram consultadas seis bases de dados eletrônicas e uma biblioteca virtual com apoio da plataforma Rayyan. Os critérios de elegibilidade foram estudos disponíveis e publicados na íntegra, que abordassem estratégias de coping no cuidado familiar a crianças com síndrome de Down. A análise dos dados foi do tipo narrativa. Resultados foram recuperados 381 registros e 14 permaneceram nesta revisão. Seis estilos de coping foram identificados: manejo das emoções, tomadas de decisão, aquisição de conhecimentos sobre a síndrome de Down e situações que a envolvem, busca de apoio, comunicação intrafamiliar e com profissionais e práticas de espiritualidade. Conclusão para equilibrar as demandas apresentadas pelas crianças e os recursos disponíveis, os integrantes da família precisam adotar estratégias cognitivas e comportamentais para lidar com as adversidades nos cuidados com a criança com síndrome de Down. Contribuições para a prática: o estudo apresenta evidências para nortear enfermeiros e outros profissionais de saúde a guiar famílias na utilização de diferentes formas de enfrentamento diante dos eventos estressores vivenciados por estas.

ABSTRACT Objective to map the coping strategies that contribute to family care for children with Down syndrome. Methods scoping review, using the JBI method. Six electronic databases and a virtual library were consulted with the support of the Rayyan platform. The criteria for eligibility were studies available and published in full, which addressed coping strategies in family care for children with Down syndrome. Data analysis was of the narrative type. Results 381 records were retrieved and 14 remained in this review. Six coping styles were identified: managing emotions, decision-making, acquiring knowledge about Down's syndrome and situations involving it, seeking support, communicating within the family and with professionals, and practicing spirituality. Conclusion in order to balance the demands presented by the children and the resources available, family members need to adopt cognitive and behavioral strategies to deal with the adversities of caring for a child with Down syndrome. Contributions to practice: the study presents evidence to guide nurses and other health professionals to help families use different ways of coping with the stressful events they experience.

Factores promotores y amenazadores de Esperanza en cuidadores de niños con condiciones crónicas / Factors that promote and threaten Hope in caregivers of children with chronic conditions / Fatores promotores e ameaçadores da Esperança em cuidadores de crianças com condições crônicas

Objetivo: identificar los factores que promueven y amenazan la Esperanza en cuidadores familiares de niños de 2 a 3 años con condiciones crónicas. Método: estudio cualitativo, incluyendo 46 cuidadores familiares de niños con condición crónica de 2 a 3 años, egresados de dos Unidades de Cuidados Intensivos Neonatales. La recolección de datos ocurrió mediante entrevistas semiestructuradas guiadas por el Modelo de Intervención en Ayuda Mutua Promotora de Esperanza. Los datos fueron sometidos a análisis temático deductivo. Resultados: se identificaron como factores promotores de la Esperanza: la experiencia compartida con los miembros de la red social de apoyo; la relación con el niño; mejoría clínica del niño; espiritualidad; orientación positiva para el futuro. Se identificaron como factores amenazadores de la Esperanza: relaciones conflictivas y incredulidad con relación al niño por parte de personas cercanas; incertidumbres sobre el futuro; inseguridades sobre la capacidad de cuidar al niño. Conclusión: los factores amenazadores de Esperanza generaron sufrimiento, dolor, angustia, ansiedad y soledad en los cuidadores. Los factores promotores de Esperanza generaron consuelo, motivación, fuerza y alegría. Los hallazgos permiten a los enfermeros reconocer las fortalezas y debilidades de los cuidadores y promover la adopción de comportamientos que promuevan la Esperanza en los cuidadores de niños con condiciones crónicas.

Objective: to identify the factors that promote and threaten Hope in family caregivers of 2- to 3-year-old children with chronic conditions. Method: qualitative study with 46 family caregivers of children between 2 and 3 years old with a chronic condition, discharged from two Neonatal Intensive Care Units. Data was collected through semi-structured interviews guided by the Model for Intervention in Mutual Help Promoter of Hope. Data were submitted to deductive thematic analysis. Results: the following were identified as factors that promote Hope: The experience shared with members of the social support network; The relationship with the child; Clinical improvement of the child; Spirituality; Positive guidance for the future. The following were identified as factors that threaten Hope: Conflictual relationships and discredit of the child by close people; Uncertainties about the future; Insecurities about the ability to care for the child. Conclusion: the threatening factors of Hope generated suffering, pain, anguish, anxiety, and loneliness in caregivers. The promoting factors of Hope generated comfort, motivation, strength and joy. The findings allow Nurses to recognize the strengths and weaknesses of caregivers and adopt behaviors that promote Hope in caregivers of children with chronic conditions.

Objetivo: identificar quais são os fatores promotores e ameaçadores da Esperança em cuidadores familiares de crianças de 2 a 3 anos com condições crônicas. Método: estudo qualitativo, incluindo 46 cuidadores familiares de crianças com condição crônica de 2 a 3 anos egressas de duas Unidades de Terapia Intensiva Neonatal. A coleta de dados se deu por meio de entrevista semiestruturada orientada pelo Modelo de Intervenção em Ajuda Mútua Promotor de Esperança. Os dados foram submetidos à análise temática dedutiva. Resultados: foram identificados como fatores promotores da Esperança: A experiência compartilhada com membros da rede de apoio social; A relação com a criança; Melhora clínica da criança; Espiritualidade; Orientação positiva para o futuro. Foram identificados como fatores ameaçadores da Esperança: Relações conflituosas e descrença da criança por pessoas próximas; Incertezas sobre o futuro; Inseguranças sobre a capacidade de prestar os cuidados à criança. Conclusão: os fatores ameaçadores da Esperança geraram sofrimento, dor, angústia, ansiedade e solidão nos cuidadores. Os fatores promotores da Esperança geraram conforto, motivação, força e alegria. Os achados possibilitam que Enfermeiros reconheçam as potencialidades e fragilidades dos cuidadores e promovam a adoção de comportamentos promotores de Esperança em cuidadores de crianças com condições crônicas.

Gestão familiar de crianças com condições crónicas: Perspetivas dos cuidadores / Family management styles of chronic childhood conditions: The caregivers' perspectives / Gestión familiar de niños con enfermedades crónicas: Perspectivas de los cuidadores

Resumo Enquadramento: Diferentes estilos de gestão podem modificar o funcionamento familiar, favorecendo ou comprometendo o cuidado das condições crónicas na infância. Objetivo: Analisar, na perspetiva do cuidador principal, a gestão familiar em situações que envolvem condições crónicas na infância. Metodologia: Estudo qualitativo, orientado pelo Family Management Style Framework, realizado a partir de entrevistas com 38 cuidadores de crianças com condições crónicas, atendidas numa instituição de saúde do sudeste brasileiro. Os dados foram submetidos a análise temática do tipo dedutiva. Resultados: A situação da condição crónica é definida como difícil e causadora de sofrimento, exigindo uma reorganização da rotina familiar para atender às exigências da criança. As consequências percebidas expressam perspetivas e receios sobre o futuro e reconhecimento dos resultados dos comportamentos de gestão adotados. foram identificados estilos de gestão como a família ajustada, em adaptação, em luta, em conflito e em espera. Conclusão: A capacidade do cuidador principal em gerenciar as necessidades da criança com condição crónica, de forma positiva, depende do acesso aos recursos profissionais, assistenciais e intrafamiliares.

Abstract Background: Different management styles can modify families' functioning, favoring or compromising the care delivered to chronic childhood conditions. Objective: To analyze primary caregivers' perspectives on family management of chronic childhood conditions. Methodology: This qualitative study is based on the Family Management Style Framework (FMSF). Thirty-eight (38) interviews were conducted with 38 caregivers of chronically ill children attending a health institution in southeastern Brazil. The data were submitted to deductive thematic analysis. Results: Having a child with chronic illness was described as challenging and causing suffering, requiring families to reorganize their family routines to meet the chronically ill child's needs. The "perceived consequences" revealed families' perspectives and fears about the future and their recognition of the results of the "management behaviors" adopted. The family management styles identified were: "thriving," "accommodating," "struggling," "floundering," and "enduring." Conclusion: The primary caregiver's ability to manage the chronically ill child's needs depends on their access to professional, care, and intra-family resources.

Resumen Marco contextual: Los diferentes estilos de gestión pueden modificar el funcionamiento familiar, lo que favorece o compromete el cuidado de las enfermedades crónicas en la infancia. Objetivo: Analizar, desde la perspectiva del cuidador principal, la gestión familiar en situaciones de afecciones crónicas en la infancia. Metodología: Estudio cualitativo, orientado por el Family Management Style Framework, realizado a partir de entrevistas a 38 cuidadores de niños con enfermedades crónicas, atendidos en una institución de salud del sudeste de Brasil. Los datos fueron sometidos a análisis temático de tipo deductivo. Resultados: La situación de la enfermedad crónica se define como difícil y causante de sufrimiento, lo que exige una reorganización de la rutina familiar para atender a las demandas del niño. Las consecuencias percibidas expresan perspectivas y temores sobre el futuro y el reconocimiento de los resultados de los comportamientos de gestión adoptados. Se identificaron como estilos de gestión: la familia ajustada, en adaptación, en lucha, en conflicto y en espera. Conclusión: La capacidad del cuidador principal para gestionar de forma positiva las necesidades del niño con una enfermedad crónica depende del acceso a recursos profesionales, asistenciales e intrafamiliares.

Chronic Intermittent Hypoxia-Induced Dysmetabolism Is Associated with Hepatic Oxidative Stress, Mitochondrial Dysfunction and Inflammation.

The association between obstructive sleep apnea (OSA) and metabolic disorders is well-established; however, the underlying mechanisms that elucidate this relationship remain incompletely understood. Since the liver is a major organ in the maintenance of metabolic homeostasis, we hypothesize that liver dysfunction plays a crucial role in the pathogenesis of metabolic dysfunction associated with obstructive sleep apnea (OSA). Herein, we explored the underlying mechanisms of this association within the liver. Experiments were performed in male Wistar rats fed with a control or high fat (HF) diet (60% lipid-rich) for 12 weeks. Half of the groups were exposed to chronic intermittent hypoxia (CIH) (30 hypoxic (5% O2) cycles, 8 h/day) that mimics OSA, in the last 15 days. Insulin sensitivity and glucose tolerance were assessed. Liver samples were collected for evaluation of lipid deposition, insulin signaling, glucose homeostasis, hypoxia, oxidative stress, antioxidant defenses, mitochondrial biogenesis and inflammation. Both the CIH and HF diet induced dysmetabolism, a state not aggravated in animals submitted to HF plus CIH. CIH aggravates hepatic lipid deposition in obese animals. Hypoxia-inducible factors levels were altered by these stimuli. CIH decreased the levels of oxidative phosphorylation complexes in both groups and the levels of SOD-1. The HF diet reduced mitochondrial density and hepatic antioxidant capacity. The CIH and HF diet produced alterations in cysteine-related thiols and pro-inflammatory markers. The results obtained suggest that hepatic mitochondrial dysfunction and oxidative stress, leading to inflammation, may be significant factors contributing to the development of dysmetabolism associated with OSA.

The Influence of Adipocyte Secretome on Selected Metabolic Fingerprints of Breast Cancer Cell Lines Representing the Four Major Breast Cancer Subtypes.

Molecular subtype (MS) is one of the most used classifications of breast cancer (BC). Four MSs are widely accepted according to receptor expression of estrogen, progesterone, and HER2. The impact of adipose tissue on BC MS metabolic impairment is still unclear. The present work aims to elucidate the metabolic alterations in breast cancer cell lines representing different MSs subjected to adipocyte associated factors. Preadipocytes isolated from human subcutaneous adipose tissue were differentiated into mature adipocytes. MS representative cell lines were exposed to mature adipocyte secretome. Extracellular medium was collected for metabolomics and RNA was extracted to evaluate enzymatic expression by RT-PCR. Adipocyte secretome exposure resulted in a decrease in the Warburg effect rate and an increase in cholesterol release. HER2+ cell lines (BT-474 and SK-BR-3) exhibited a similar metabolic pattern, in contrast to luminal A (MCF-7) and triple negative (TN) (MDA-MB-231), both presenting identical metabolisms. Anaplerosis was found in luminal A and TN representative cells, whereas cataplerotic reactions were likely to occur in HER2+ cell lines. Our results indicate that adipocyte secretome affects the central metabolism distinctly in each BC MS representative cell line.

Simultaneous determination of nine high-intensity sweeteners in liquid and powder tabletop sweeteners.

Government regulatory actions and public policies have been recently implemented in Brazil due to the excessive consumption of sugar. Therefore, it becomes relevant to determine the levels of high-intensity sweeteners in tabletop sweeteners consumed by the Brazilian population. Thus, an analytical method was developed and validated for the simultaneous determination of nine sweeteners (acesulfame potassium, aspartame, advantame, sodium cyclamate, neotame, saccharin, sucralose, stevioside, and rebaudioside A) by using ultra-high performance liquid chromatography coupled to mass spectrometry in tandem. The sample preparation encompassed only dilution steps. The method was validated taking into account the parameters of linearity, precision, accuracy, and matrix effects. The analytes were determined in two different batches of 21 commercial liquid and powder tabletop sweeteners available on the local market, totaling 42 samples. A minimum of one and a maximum of four sweeteners were found in the analyzed products and sweeteners that were not described on the label were not detected. It is expected that the established method can be used in monitoring programs and that the presented results can contribute to exposure assessments performed nationally.

The progression from mild to severe hyperglycemia coupled with insulin resistance causes mitochondrial dysfunction and alters the metabolic secretome of epithelial kidney cells.

Diabetic nephropathy (DN) and insulin resistance (IR) in kidney cells are considered main causes for end-stage renal failure. However, it is unclear how IR affects early stages of the disease. Here, we investigate the impact of mild (11 mM) and severe (22 mM) hyperglycemia, with and without induced IR, on cellular metabolism and mitochondrial bioenergetics in a human kidney cell line (HK-2). IR in HK-2 cells was induced with palmitic acid and cellular cytotoxicity was studied. We evaluated the impact of mild and severe hyperglycemia with and without IR on the metabolic secretome of the cells, their live-cell mitochondria function, mitochondrial membrane potential, and mitochondrial complex activities. Furthermore, we measured fatty acid oxidation and lipid accumulation. Cells cultured under mild hyperglycemic conditions exhibited increased mitochondrial bioenergetic parameters, such as basal respiration, ATP-linked production, maximal respiration capacity, and spare respiration capacity. However, these parameters decreased when cells were cultured under higher glucose concentrations when IR was induced. Our data suggests that progression from mild to severe hyperglycemia induces a metabolic shift, where gluconeogenic amino acids play a crucial role in supplying the energy requirements of HK-2. To our knowledge, this is the first study to evaluate the progression from mild to severe hyperglycemia allied to IR in human kidney cells. This work highlights that this progression leads to mitochondrial dysfunction and alters the metabolic profile of kidney cells. These results identify possible targets for early intervention in DN.

Differentiation of peripheral sensory neurons from iPSCs derived from stem cells from human exfoliated deciduous teeth (SHED).

Peripheral nervous system (PNS) sensory alterations are present in several pathologies and syndromes. The use of induced pluripotent stem cell (iPSC) technology is an important strategy to produce sensory neurons in patients who are accomplished in terms of sensory symptoms. The iPSC technology relies on manipulating signaling pathways to resemble what occurs in vivo, and the iPSCs are known to carry a transcriptional memory after reprogramming, which can affect the produced cell. To this date, protocols described for sensory neuron production start using iPSCs derived from skin fibroblasts, which have the same ontogenetic origin as the central nervous system (CNS). Since it is already known that the cells somehow resemble their origin even after cell reprogramming, PNS cells should be produced from cells derived from the neural crest. This work aimed to establish a protocol to differentiate sensory neurons derived from stem cells from human exfoliated deciduous teeth (SHED) with the same embryonic origin as the PNS. SHED-derived iPSCs were produced and submitted to peripheral sensory neuron (PSN) differentiation. Our protocol used the dual-SMAD inhibition method, followed by neuronal differentiation, using artificial neurotrophic factors and molecules produced by human keratinocytes. We successfully established the first protocol for differentiating neural crest and PNS cells from SHED-derived iPSCs, enabling future studies of PNS pathologies.

Mitochondria Quality Control and Male Fertility.

Mitochondria are pivotal to cellular homeostasis, performing vital functions such as bioenergetics, biosynthesis, and cell signalling. Proper maintenance of these processes is crucial to prevent disease development and ensure optimal cell function. Mitochondrial dynamics, including fission, fusion, biogenesis, mitophagy, and apoptosis, maintain mitochondrial quality control, which is essential for overall cell health. In male reproduction, mitochondria play a pivotal role in germ cell development and any defects in mitochondrial quality can have serious consequences on male fertility. Reactive oxygen species (ROS) also play a crucial role in sperm capacitation, but excessive ROS levels can trigger oxidative damage. Any imbalance between ROS and sperm quality control, caused by non-communicable diseases or environmental factors, can lead to an increase in oxidative stress, cell damage, and apoptosis, which in turn affect sperm concentration, quality, and motility. Therefore, assessing mitochondrial functionality and quality control is essential to gain valuable insights into male infertility. In sum, proper mitochondrial functionality is essential for overall health, and particularly important for male fertility. The assessment of mitochondrial functionality and quality control can provide crucial information for the study and management of male infertility and may lead to the development of new strategies for its management.

Overview of SARS-COV-2 infection at the Butantan Penitentiary Progression Center.

OBJECTIVE: To estimate the prevalence of exposure to the SARS-CoV-2 virus among individuals living in restricted freedom. METHODS: A seroprevalence survey was carried out with the population of the female penitentiary of the Centro de Progressão Penitenciária (CPP) in Butantan (municipality of São Paulo), between June 24 and August 20, 2020. During this period, according to the Secretariat of Penitentiary Administration (SAP), the positivity of rapid tests among inmates ranged from 65% to 78%. The evaluation method used in the study was the "One Step COVID-19" rapid test (chromatography), from the company Wondfo, also using the RT-PCR method in symptomatic participants to confirm the viral condition. The study population consisted of 879 female inmates and 170 employees of the institution. RESULTS: The prevalence of total antibodies (IgG/IgM) against the SARS-CoV-2 virus in the total population of 1049 study participants was 6.1%; among the population of 879 inmates,a prevalence of 5.8% was observed, and among the institution's employees, 7.5%. CONCLUSIONS: The prevalence of covid-19 at the Butantan CPP was low, which is due to the implementation of simple prevention measures at the institution, such as the use of masks (with appropriate changes), emphasis on hygiene, hand washing and social distancing, in addition to other strategies, such as suspending inmates' visits from relatives and friends and cutting back on elective medical appointments and outside work.

Renal Dysfunction Phenotypes in Patients Undergoing Obesity Surgery.

Obesity surgery candidates are at an increased risk of kidney injury, but pre-operative evaluation usually neglects kidney function assessment. This study aimed to identify renal dysfunction in candidates for bariatric surgery. To reduce the sources of bias, subjects with diabetes, prediabetes under metformin treatment, neoplastic or inflammatory diseases were excluded. Patients' (n = 192) average body mass index was 41.7 ± 5.4 kg/m2. Among these, 51% (n = 94) had creatinine clearance over 140 mL/min, 22.4% (n = 43) had proteinuria over 150 mg/day and 14.6% (n = 28) albuminuria over 30 mg/day. A creatinine clearance higher than 140 mL/min was associated with higher levels of proteinuria and albuminuria. Univariate analysis identified sex, glycated hemoglobin, uric acid, HDL and VLDL cholesterol as being associated with albuminuria, but not with proteinuria. On multivariate analysis, glycated hemoglobin and creatinine clearance as continuous variables were significantly associated with albuminuria. In summary, in our patient population prediabetes, lipid abnormalities and hyperuricemia were associated with albuminuria, but not with proteinuria, suggesting different disease mechanisms might be implicated. Data suggest that in obesity-associated kidney disease, tubulointerstitial injury precedes glomerulopathy. A significant proportion of obesity surgery candidates present clinically relevant albuminuria and proteinuria along with renal hyperfiltration, suggesting that routine pre-operative assessment of these parameters should be considered.

Factors that promote and threaten Hope in caregivers of children with chronic conditions. / Factores promotores y amenazadores de Esperanza en cuidadores de niños con condiciones crónicas.

to identify the factors that promote and threaten Hope in family caregivers of 2- to 3-year-old children with chronic conditions. qualitative study with 46 family caregivers of children between 2 and 3 years old with a chronic condition, discharged from two Neonatal Intensive Care Units. Data was collected through semi-structured interviews guided by the Model for Intervention in Mutual Help Promoter of Hope. Data were submitted to deductive thematic analysis. the following were identified as factors that promote Hope: The experience shared with members of the social support network; The relationship with the child; Clinical improvement of the child; Spirituality; Positive guidance for the future. The following were identified as factors that threaten Hope: Conflictual relationships and discredit of the child by close people; Uncertainties about the future; Insecurities about the ability to care for the child. the threatening factors of Hope generated suffering, pain, anguish, anxiety, and loneliness in caregivers. The promoting factors of Hope generated comfort, motivation, strength and joy. The findings allow Nurses to recognize the strengths and weaknesses of caregivers and adopt behaviors that promote Hope in caregivers of children with chronic conditions.

identificar los factores que promueven y amenazan la Esperanza en cuidadores familiares de niños de 2 a 3 años con condiciones crónicas. estudio cualitativo, incluyendo 46 cuidadores familiares de niños con condición crónica de 2 a 3 años, egresados de dos Unidades de Cuidados Intensivos Neonatales. La recolección de datos ocurrió mediante entrevistas semiestructuradas guiadas por el Modelo de Intervención en Ayuda Mutua Promotora de Esperanza. Los datos fueron sometidos a análisis temático deductivo. se identificaron como factores promotores de la Esperanza: la experiencia compartida con los miembros de la red social de apoyo; la relación con el niño; mejoría clínica del niño; espiritualidad; orientación positiva para el futuro. Se identificaron como factores amenazadores de la Esperanza: relaciones conflictivas y incredulidad con relación al niño por parte de personas cercanas; incertidumbres sobre el futuro; inseguridades sobre la capacidad de cuidar al niño. los factores amenazadores de Esperanza generaron sufrimiento, dolor, angustia, ansiedad y soledad en los cuidadores. Los factores promotores de Esperanza generaron consuelo, motivación, fuerza y alegría. Los hallazgos permiten a los enfermeros reconocer las fortalezas y debilidades de los cuidadores y promover la adopción de comportamientos que promuevan la Esperanza en los cuidadores de niños con condiciones crónicas.

(1) La esperanza fortalece a las familias de niños con condiciones crónicas. (2) Hay factores capaces de promover o amenazar la Esperanza de los padres. (3)Los factores que promueven la esperanza generan comodidad, motivación, fuerza y alegría. (4) Los factores amenazadores de la esperanza generan sufrimiento, angustia, ansiedad y soledad. (5) Los enfermeros pueden adoptar comportamientos que promuevan la esperanza y ayudar a las familias.

identificar quais são os fatores promotores e ameaçadores da Esperança em cuidadores familiares de crianças de 2 a 3 anos com condições crônicas. estudo qualitativo, incluindo 46 cuidadores familiares de crianças com condição crônica de 2 a 3 anos egressas de duas Unidades de Terapia Intensiva Neonatal. A coleta de dados se deu por meio de entrevista semiestruturada orientada pelo Modelo de Intervenção em Ajuda Mútua Promotor de Esperança. Os dados foram submetidos à análise temática dedutiva. foram identificados como fatores promotores da Esperança: A experiência compartilhada com membros da rede de apoio social; A relação com a criança; Melhora clínica da criança; Espiritualidade; Orientação positiva para o futuro. Foram identificados como fatores ameaçadores da Esperança: Relações conflituosas e descrença da criança por pessoas próximas; Incertezas sobre o futuro; Inseguranças sobre a capacidade de prestar os cuidados à criança. os fatores ameaçadores da Esperança geraram sofrimento, dor, angústia, ansiedade e solidão nos cuidadores. Os fatores promotores da Esperança geraram conforto, motivação, força e alegria. Os achados possibilitam que Enfermeiros reconheçam as potencialidades e fragilidades dos cuidadores e promovam a adoção de comportamentos promotores de Esperança em cuidadores de crianças com condições crônicas.

The role of ion homeostasis imbalance due to citrate accumulation in fluoroacetic acid (FAA) toxicity in Neurospora crassa.

Fluoroacetic acid (FAA) is a poison commonly used for the lethal control of invasive species in Australia and New Zealand. Despite its widespread use and long history as a pesticide, no effective treatment for accidental poisoning exists. Although it is known to inhibit the tricarboxylic acid (TCA) cycle, specific details of FAA toxicology have remained elusive, with hypocalcemia suggested to be involved in the neurological symptoms prior to death. Here, we study the effects of FAA on cell growth and mitochondrial function using the filamentous fungi Neurospora crassa as model organism. FAA toxicosis in N. crassa is characterized by an initial hyperpolarization and subsequent depolarization of the mitochondrial membranes, followed by a significant intracellular decrease in ATP and increase in Ca2+. The development of mycelium was markedly affected within 6 h, and growth impaired after 24 h of FAA exposure. Although the activity of mitochondrial complexes I, II and IV was impaired, the activity of citrate synthase was not affected. Supplementation with Ca2+ exacerbated the effects of FAA in cell growth and membrane potential. Our findings suggest that an imbalance created in the ratio of ions within the mitochondria may lead to conformational changes in ATP synthase dimers due to mitochondrial Ca2+ uptake, that ultimately result in the opening of the mitochondrial permeability transition pore (MPTP), a decrease in membrane potential, and cell death. Our findings suggest new approaches for the treatment research, as well as the possibility to use N. crassa as a high-throughput screening assay to evaluate a large number of FAA antidote candidates.