Contribution of extracellular vesicles for the pathogenesis of retinal diseases: shedding light on blood-retinal barrier dysfunction.

Retinal degenerative diseases, including diabetic retinopathy (DR) and age-related macular degeneration (AMD), loom as threats to vision, causing detrimental effects on the structure and function of the retina. Central to understanding these diseases, is the compromised state of the blood-retinal barrier (BRB), an effective barrier that regulates the influx of immune and inflammatory components. Whether BRB breakdown initiates retinal distress, or is a consequence of disease progression, remains enigmatic. Nevertheless, it is an indication of retinal dysfunction and potential vision loss.The intricate intercellular dialogues among retinal cell populations remain unintelligible in the complex retinal milieu, under conditions of inflammation and oxidative stress. The retina, a specialized neural tissue, sustains a ceaseless demand for oxygen and nutrients from two vascular networks. The BRB orchestrates the exchange of molecules and fluids within this specialized region, comprising the inner BRB (iBRB) and the outer BRB (oBRB). Extracellular vesicles (EVs) are small membranous structures, and act as messengers facilitating intercellular communication in this milieu.EVs, both from retinal and peripheral immune cells, increase complexity to BRB dysfunction in DR and AMD. Laden with bioactive cargoes, these EVs can modulate the retinal microenvironment, influencing disease progression. Our review delves into the multifaceted role of EVs in retinal degenerative diseases, elucidating the molecular crosstalk they orchestrate, and their microRNA (miRNA) content. By shedding light on these nanoscale messengers, from their biogenesis, release, to interaction and uptake by target cells, we aim to deepen the comprehension of BRB dysfunction and explore their therapeutic potential, therefore increasing our understanding of DR and AMD pathophysiology.

Molecular Pharmacology in Diabetes.

This Special Issue highlights the key molecules and molecular signaling pathways associated with diabetes and its multifaceted complications [...].

Evaluation of patients' satisfaction with domiciliary biological treatment in severe asthma: a Portuguese survey.

Dear Editor, According to GINA, severe asthma is defined as uncontrolled asthma, despite therapy adherence with an optimized high dose of inhaled corticosteroid plus long-acting ß2-agonist coupled with management of modifiable factors/comorbidities, that worsens when this treatment is decreased. It affects a significant portion of asthmatic patients and imposes a high risk of exacerbations and mortality, which are associated with significant healthcare costs and psychosocial impact...

Disturbed Matrix Metalloproteinases Activity in Age-Related Macular Degeneration.

Matrix metalloproteinases (MMPs) are a tightly regulated family of proteolytic enzymes that break down extracellular matrix (ECM) and basement membrane components. Because it is associated with development, morphogenesis, tissue remodeling, and repair, ECM remodeling is an important mechanism. MMPs are thought to act as a double-edged sword, as they contribute to maintaining photoreceptors/retinal pigment epithelium (RPE)/Bruch's membrane (BM)/choroid complex homeostasis and also contribute to the onset and progression of age-related macular degeneration (AMD). Polymorphisms and/or altered expression in MMPs and their tissue inhibitors (TIMPs) are associated with age-related macular degeneration (AMD). Here, we review the evidence for MMPs' role in the onset and progression of AMD via addressing their regulation and TIMPs' significant regulatory functions.

Somatic mutations of CADM1 in aldosterone-producing adenomas and gap junction-dependent regulation of aldosterone production.

Aldosterone-producing adenomas (APAs) are the commonest curable cause of hypertension. Most have gain-of-function somatic mutations of ion channels or transporters. Herein we report the discovery, replication and phenotype of mutations in the neuronal cell adhesion gene CADM1. Independent whole exome sequencing of 40 and 81 APAs found intramembranous p.Val380Asp or p.Gly379Asp variants in two patients whose hypertension and periodic primary aldosteronism were cured by adrenalectomy. Replication identified two more APAs with each variant (total, n = 6). The most upregulated gene (10- to 25-fold) in human adrenocortical H295R cells transduced with the mutations (compared to wildtype) was CYP11B2 (aldosterone synthase), and biological rhythms were the most differentially expressed process. CADM1 knockdown or mutation inhibited gap junction (GJ)-permeable dye transfer. GJ blockade by Gap27 increased CYP11B2 similarly to CADM1 mutation. Human adrenal zona glomerulosa (ZG) expression of GJA1 (the main GJ protein) was patchy, and annular GJs (sequelae of GJ communication) were less prominent in CYP11B2-positive micronodules than adjacent ZG. Somatic mutations of CADM1 cause reversible hypertension and reveal a role for GJ communication in suppressing physiological aldosterone production.

Immune Fingerprint in Diabetes: Ocular Surface and Retinal Inflammation.

Diabetes is a prevalent global health issue associated with significant morbidity and mortality. Diabetic retinopathy (DR) is a well-known inflammatory, neurovascular complication of diabetes and a leading cause of preventable blindness in developed countries among working-age adults. However, the ocular surface components of diabetic eyes are also at risk of damage due to uncontrolled diabetes, which is often overlooked. Inflammatory changes in the corneas of diabetic patients indicate that inflammation plays a significant role in diabetic complications, much like in DR. The eye's immune privilege restricts immune and inflammatory responses, and the cornea and retina have a complex network of innate immune cells that maintain immune homeostasis. Nevertheless, low-grade inflammation in diabetes contributes to immune dysregulation. This article aims to provide an overview and discussion of how diabetes affects the ocular immune system's main components, immune-competent cells, and inflammatory mediators. By understanding these effects, potential interventions and treatments may be developed to improve the ocular health of diabetic patients.

Attention-Deficit/Hyperactivity Disorder Animal Model Presents Retinal Alterations and Methylphenidate Has a Differential Effect in ADHD versus Control Conditions.

Attention-Deficit/Hyperactivity Disorder (ADHD) is one of the most prevalent neurodevelopmental disorders. Interestingly, children with ADHD seem to experience more ophthalmologic abnormalities, and the impact of methylphenidate (MPH) use on retinal physiology remains unclear. Thus, we aimed to unravel the retina's structural, functional, and cellular alterations and the impact of MPH in ADHD versus the control conditions. For that, spontaneously hypertensive rats (SHR) and Wistar Kyoto rats (WKY) were used as animal models of ADHD and the controls, respectively. Animals were divided into four experimental groups as follows: WKY vehicle (Veh; tap water), WKY MPH (1.5 mg/kg/day), SHR Veh, SHR MPH. Individual administration was performed by gavage between P28-P55. Retinal physiology and structure were evaluated at P56 followed by tissue collection and analysis. The ADHD animal model presents the retinal structural, functional, and neuronal deficits, as well as the microglial reactivity, astrogliosis, blood-retinal barrier (BRB) hyperpermeability and a pro-inflammatory status. In this model, MPH had a beneficial effect on reducing microgliosis, BRB dysfunction, and inflammatory response, but did not correct the neuronal and functional alterations in the retina. Curiously, in the control animals, MPH showed an opposite effect since it impaired the retinal function, neuronal cells, and BRB integrity, and also promoted both microglia reactivity and upregulation of pro-inflammatory mediators. This study unveils the retinal alterations in ADHD and the opposite effects induced by MPH in the retina of ADHD and the control animal models.

TRAP1 Is Expressed in Human Retinal Pigment Epithelial Cells and Is Required to Maintain their Energetic Status.

Age-related macular degeneration (AMD) is the leading cause of severe vision loss and blindness in elderly people worldwide. The damage to the retinal pigment epithelium (RPE) triggered by oxidative stress plays a central role in the onset and progression of AMD and results from the excessive accumulation of reactive oxygen species (ROS) produced mainly by mitochondria. Tumor necrosis factor receptor-associated protein 1 (TRAP1) is a mitochondrial molecular chaperone that contributes to the maintenance of mitochondrial integrity by decreasing the production and accumulation of ROS. The present study aimed to evaluate the presence and the role of TRAP1 in the RPE. Here, we report that TRAP1 is expressed in human adult retinal pigment epithelial cells and is located mainly in the mitochondria. Exposure of RPE cells to hydrogen peroxide decreases the levels of TRAP1. Furthermore, TRAP1 silencing increases intracellular ROS production and decreases mitochondrial respiratory capacity without affecting cell proliferation. Together, these findings offer novel insights into TRAP1 functions in RPE cells, opening possibilities to develop new treatment options for AMD.

New advances in endocrine hypertension: from genes to biomarkers.

Hypertension (HT) is a major cardiovascular risk factor that affects 10% to 40% of the general population in an age-dependent manner. Detection of secondary forms of HT is particularly important because it allows the targeted management of the underlying disease. Among hypertensive patients, the prevalence of endocrine HT reaches up to 10%. Adrenal diseases are the most frequent cause of endocrine HT and are associated with excess production of mineralocorticoids (mainly primary aldosteronism), glucocorticoids (Cushing syndrome), and catecholamines (pheochromocytoma). In addition, a few rare diseases directly affecting the action of mineralocorticoids and glucocorticoids in the kidney also lead to endocrine HT. Over the past years, genomic and genetic studies have allowed improving our knowledge on the molecular mechanisms of endocrine HT. Those discoveries have opened new opportunities to transfer knowledge to clinical practice for better diagnosis and specific treatment of affected subjects. In this review, we describe the physiology of adrenal hormone biosynthesis and action, the clinical and biochemical characteristics of different forms of endocrine HT, and their underlying genetic defects. We discuss the impact of these discoveries on diagnosis and management of patients, as well as new perspectives related to the use of new biomarkers for improved patient care.

Os quilombos e suas interfaces com o Sistema Único de Assistência Social / Quilombos and their interfaces with the Unified Social Assistance System

O artigo apresenta a discussão sobre os quilombos no estado do Rio Grande do Sul (RS) e as aproximações com as ofertas inscritas no Sistema Único de Assistência Social (SUAS), com objetivo de analisar as particularidades da questão étnico-racial acerca dos povos quilombolas no estado do Rio Grande do Sul, para a afirmação desta pauta na agenda do SUAS. Socializa os resultados de uma pesquisa documental de abordagem mista com ênfase qualitativa. Revela a potência e a complementaridade das informações encontradas nos instrumentos do SUAS, como nos dados do Censo SUAS (2019) do Centro de Referência de Assistência Social (CRAS) e do Centro de Referência Especializado de Assistência Social (CREAS) e os dados do Cadastro Único (2021), frente ao reconhecimento dos territórios quilombolas, embora ainda seja necessário ampliar as ações de educação permanente acerca do debate sobre a questão étnico-racial e os quilombos na agenda da política de assistência social

The article presents the discussion about the quilombos in the state of Rio Grande do Sul (RS) and the approximations with the offers registered in the Unified Social Assistance System (SUAS), with the objective of analyzing the particularities of the ethnic-racial issue concerning the quilombola peoples in the state of Rio Grande do Sul, for the affirmation of this agenda in the SUAS agenda. It socializes the results of a mixed approach documentary research with a qualitative emphasis. It reveals the power and complementarity of the information found in the SUAS instruments, as in the data from the SUAS Census (2019) from the Social Assistance Reference Center (CRAS) and the Specialized Reference Center for Social Assistance (CREAS) and data from the Single Registry (2021), in view of the recognition of quilombola territories, although it is still necessary to expand permanent education actions regarding the debate on the ethnic-racial issue and the quilombos in the social assistance policy agenda

Adaptação e validação de Índice de Qualidade de Vida para gestantes brasileiras / Adaptation and validity of the Quality of Life Index for Brazilian pregnant women / Adaptation and validity of the Quality of Life Index for Brazilian pregnant women / Adaptación y validación del Índice de Calidad de Vida para mujeres embarazadas brasileñas

Resumo Objetivo Adaptar e validar o Índice de Qualidade de Vida de Ferrans & Powers para gestantes brasileiras. Métodos Estudo metodológico. A versão brasileira Índice de Qualidade de Vida de Ferrans & Powers para gestantes tem 36 itens e quatro domínios. A validação de conteúdo foi realizada por comitê de juízes. Na etapa de validação foi testada a consistência interna, a validade de constructo convergente e discriminante e a dimensionalidade. Nível de significância 5%. Resultados Cinco juízes participaram do comitê. O índice de validade de conteúdo foi de 0,94 e a maioria dos itens apresentou coeficiente de validade de conteúdo por item acima de 0,80. Participaram da etapa de validação 280 gestantes.O alfa de Cronbach foi de 0,94 para o escore total com variação de 0,78 a 0,89 entre os domínios A correlação de Pearson entre o Índice de Qualidade de Vida de Ferrans e o WHOQOL-Bref foi positiva e forte (0,79; p<0,001). A validade de constructo discriminante não revelou diferenças estatisticamente significante. A análise fatorial confirmatória revelou que o modelo de quatro domínios se ajusta ao modelo. Conclusão A versão adaptada do Índice de Qualidade de Vida de Ferrans mostrou-se confiável e válida para aplicação em gestantes, mostrando-se uma ferramenta promissora para profissionais de saúde e pesquisadores na identificação da Qualidade de vida de gestantes.

Resumen Objetivo Adaptar y validar el Índice de Calidad de Vida de Ferrans & Powers para mujeres embarazadas brasileñas. Métodos Estudio metodológico. La versión brasileña del Índice de Calidad de Vida de Ferrans & Powers para mujeres embarazadas tiene 36 ítems y cuatro dominios. La validación de contenido fue realizada por un comité de jueces. En la etapa de validación se probó la consistencia interna, la validez de constructo convergente y discriminante y la dimensionalidad. Nivel de significación del 5 %. Resultados Cinco jueces participaron del comité. El índice de validez de contenido fue de 0,94 y la mayoría de los ítems presentó un coeficiente de validez de contenido por ítem superior a 0,80. En la etapa de validación participaron 280 mujeres embarazadas. El alfa de Cronbach fue de 0,94 para el puntaje total con variación de 0,78 a 0,89 entre los dominios. La correlación de Pearson entre el Índice de Calidad de Vida de Ferrans y el WHOQOL-Bref fue positiva y fuerte (0,79; p<0,001). La validez del constructo discriminante no reveló diferencias estadísticamente significativas. El análisis factorial confirmatorio reveló que el modelo de cuatro dominios se ajusta al modelo. Conclusión La versión adaptada del Índice de Calidad de Vida de Ferrans demostró ser confiable y válida para su uso en mujeres embarazadas y demostró ser una herramienta promisora para profesionales de la salud y para investigadores en la identificación de calidad de vida de mujeres embarazadas.

Abstract Objective To adapt and validate the Ferrans and Powers Quality of Life Index for Brazilian pregnant women. Methods This is a methodological study. The Ferrans and Powers Quality of Life Index for pregnant women, Brazilian version, has 36 items and four domains. Content validity was performed by a committee of judges. In the validity stage, internal consistency, convergent and discriminant construct validity and dimensionality were tested. Significance level 5%. Results Five judges participated in a committee. The Content Validity Index was 0.94 and most items had a content validity coefficient per item above 0.80. A total of 280 pregnant women participated in the validity stage. Cronbach's alpha was 0.94 for the total score, ranging from 0.78 to 0.89 between the domains. Pearson's correlation between the Ferrans and Powers Quality of Life Index and the WHOQOL-Bref was positive and strong (0.79; p<0.001). Discriminant construct validity did not reveal statistically significant differences. Confirmatory factor analysis revealed that the four-domain model fits the model. Conclusion The Ferrans and Powers Quality of Life Index, adapted version, proved to be reliable and valid for use in pregnant women, proving to be a promising tool for health professionals and researchers to identify pregnant women's quality of life.

Vascular and hormonal interactions in the adrenal gland.

Primary aldosteronism is the most common form of secondary arterial hypertension, due to excessive aldosterone production from the adrenal gland. Although somatic mutations have been identified in aldosterone producing adenoma, the exact mechanisms leading to increased cell proliferation and nodule formation remain to be established. One hypothesis is that changes in vascular supply to the adrenal cortex, due to phenomena of atherosclerosis or high blood pressure, may influence the morphology of the adrenal cortex, resulting in a compensatory growth and nodule formation in response to local hypoxia. In this review, we will summarize our knowledge on the mechanisms regulating adrenal cortex development and function, describe adrenal vascularization in normal and pathological conditions and address the mechanisms allowing the cross-talk between the hormonal and vascular components to allow the extreme tissue plasticity of the adrenal cortex in response to endogenous and exogenous stimuli. We will then address recent evidence suggesting a role for alterations in the vascular compartment that could eventually be involved in nodule formation and the development of primary aldosteronism.

Identification of risk loci for primary aldosteronism in genome-wide association studies.

Primary aldosteronism affects up to 10% of hypertensive patients and is responsible for treatment resistance and increased cardiovascular risk. Here we perform a genome-wide association study in a discovery cohort of 562 cases and 950 controls and identify three main loci on chromosomes 1, 13 and X; associations on chromosome 1 and 13 are replicated in a second cohort and confirmed by a meta-analysis involving 1162 cases and 3296 controls. The association on chromosome 13 is specific to men and stronger in bilateral adrenal hyperplasia than aldosterone producing adenoma. Candidate genes located within the two loci, CASZ1 and RXFP2, are expressed in human and mouse adrenals in different cell clusters. Their overexpression in adrenocortical cells suppresses mineralocorticoid output under basal and stimulated conditions, without affecting cortisol biosynthesis. Our study identifies the first risk loci for primary aldosteronism and highlights new mechanisms for the development of aldosterone excess.

Putative Biomarkers in Tears for Diabetic Retinopathy Diagnosis.

Purpose: Tear fluid biomarkers may offer a non-invasive strategy for detecting diabetic patients with increased risk of developing diabetic retinopathy (DR) or increased disease progression, thus helping both improving diagnostic accuracy and understanding the pathophysiology of the disease. Here, we assessed the tear fluid of nondiabetic individuals, diabetic patients with no DR, and diabetic patients with nonproliferative DR (NPDR) or with proliferative DR (PDR) to find putative biomarkers for the diagnosis and staging of DR. Methods: Tear fluid samples were collected using Schirmer test strips from a cohort with 12 controls and 54 Type 2 Diabetes (T2D) patients, and then analyzed using mass spectrometry (MS)-based shotgun proteomics and bead-based multiplex assay. Tear fluid-derived small extracellular vesicles (EVs) were analyzed by transmission electron microscopy, Western Blotting, and nano tracking. Results: Proteomics analysis revealed that among the 682 reliably quantified proteins in tear fluid, 42 and 26 were differentially expressed in NPDR and PDR, respectively, comparing to the control group. Data are available via ProteomeXchange with identifier PXD033101. By multicomparison analyses, we also found significant changes in 32 proteins. Gene ontology (GO) annotations showed that most of these proteins are associated with oxidative stress and small EVs. Indeed, we also found that tear fluid is particularly enriched in small EVs. T2D patients with NPDR have higher IL-2/-5/-18, TNF, MMP-2/-3/-9 concentrations than the controls. In the PDR group, IL-5/-18 and MMP-3/-9 concentrations were significantly higher, whereas IL-13 was lower, compared to the controls. Conclusions: Overall, the results show alterations in tear fluid proteins profile in diabetic patients with retinopathy. Promising candidate biomarkers identified need to be validated in a large sample cohort.

Improvement of Glycaemia and Endothelial Function by a New Low-Dose Curcuminoid in an Animal Model of Type 2 Diabetes.

Curcumin has been suggested as a promising treatment for metabolic diseases, but the high doses required limit its therapeutic use. In this study, a new curcuminoid is synthesised to increase curcumin anti-inflammatory and antioxidant potential and to achieve hypoglycaemic and protective vascular effects in type 2 diabetic rats in a lower dose. In vitro, the anti-inflammatory effect was determined through the Griess reaction, and the antioxidant activity through ABTS and TBARS assays. In vivo, Goto-Kakizaki rats were treated for 2 weeks with the equimolar dose of curcumin (40 mg/kg/day) or curcuminoid (52.4 mg/kg/day). Fasting glycaemia, insulin tolerance, plasma insulin, insulin signalling, serum FFA, endothelial function and several markers of oxidative stress were evaluated. Both compounds presented a significant anti-inflammatory effect. Moreover, the curcuminoid had a marked hypoglycaemic effect, accompanied by higher GLUT4 levels in adipose tissue. Both compounds increased NO-dependent vasorelaxation, but only the curcuminoid exacerbated the response to ascorbic acid, consistent with a higher decrease in vascular oxidative and nitrosative stress. SOD1 and GLO1 levels were increased in EAT and heart, respectively. Altogether, these data suggest that the curcuminoid developed here has more pronounced effects than curcumin in low doses, improving the oxidative stress, endothelial function and glycaemic profile in type 2 diabetes.

Lab-on-a-chip technologies for minimally invasive molecular sensing of diabetic retinopathy.

Diabetic retinopathy (DR) is the most common diabetic eye disease and the worldwide leading cause of vision loss in working-age adults. It progresses from mild to severe non-proliferative or proliferative DR based on several pathological features including the magnitude of blood-retinal barrier breakdown and neovascularization. Available pharmacological and retinal laser photocoagulation interventions are mostly applied in the advanced stages of DR and are inefficient in halting disease progression in a significantly high percentage of patients. Yet, recent evidence has shown that some therapies could potentially limit DR progression if applied at early stages, highlighting the importance of early disease diagnostics. In the past few decades, different imaging modalities have proved their utility for examining retinal and optic nerve changes in patients with retinal diseases. However, imaging based-methodologies solely rely on morphological examination of the retinal vascularization and are not suitable for recurrent and personalized patient evaluation. This raises the need for new technologies to enable accurate and early diagnosis of DR. In this review, we critically discuss the potential clinical benefit of minimally-invasive molecular biomarker identification and profiling of diabetic patients who are at risk of developing DR. We provide a comparative overview of conventional and recently developed lab-on-a-chip technologies for quantitative assessment of potential DR molecular biomarkers and discuss their advantages, current limitations and challenges for future practical implementation and continuous patient monitoring at the point-of-care.

Posttraumatic stress disorder in burn patient: A systematic review.

INTRODUCTION: Burns often cause severe physical and mental suffering and can become a trigger for the development of permanent psychological diseases, even after wound healing. Posttraumatic stress disorder (PTSD) is one such disorder, which involves the re-experiencing of many symptoms provoked by a previous traumatic situation. METHOD: This study is a systematic review of interventions used to reduce or prevent PTSD symptoms in burn victims. We included randomized clinical trials that described therapeutic interventions for the prevention of PTSD in burn patients. The search was conducted in the databases EMBASE and PUBMED/Medline between 2009 and 2020, and the main variables evaluated were%TBSA, age, number of hospitalization days, type of intervention, follow-up time and results. The analysis of the bias risk was carried out according to the guidance in the Cochrane Handbook for Bias Risk Assessment. RESULTS: Eight clinical trials were selected: three of them were performed in children, and five involved adults. The most common bias risks were related to participant/researcher blinding and loss of follow-up. Two interventions were identified: a pharmacological intervention and a psychological intervention. Medications (sertraline and propanolol) were not effective in reducing stress symptoms. Four studies used cognitive-behavioral therapies, which achieved the best results for PTSD improvement in burn patients. Hypnosis and an informational education program were also evaluated and did not show success in reducing PTSD. CONCLUSION: Cognitive-behavioral therapies may work to reduce PTSD symptoms in burn patients, and when they are adopted early by burn units, they may improve the psychological condition of burn patients.

Spherical and rod shaped mesoporous silica nanoparticles for cancer-targeted and photosensitizer delivery in photodynamic therapy.

Mesoporous silica nanoparticles (MSNPs) have attracted much attention in many biomedical applications. One of the fields in which smart functional nanosystems have found wide application is cancer treatment. Here, we present new silica nanoparticle-based systems which have been explored as efficient vehicles to transport and deliver photosensitizers (PSs) into tumor tissues during photodynamic therapy (PDT). In this work, we report the preparation, characterization, and in vitro studies of distinct shaped MSNPs grafted with S-glycoside porphyrins (Pors). The ensuing nanomaterials were fully characterized, and their properties as third-generation PSs for PDT against two bladder cancer cell lines, HT-1376 and UM-UC-3, were examined. The best uptake results were obtained for MSNP-PS2, while MSNP-PS1 showed the lowest cellular uptake among the nanocarriers tested, but revealed the best phototoxicity in both cancer cells. Overall, the phototoxicity was higher with MSNPs than with mesoporous silica nanorods (MSNRs) and higher uptake and phototoxicity were consistently observed in UM-UC-3 rather than in HT-1376 cancer cells.

Colocalization of Wnt/ß-Catenin and ACTH Signaling Pathways and Paracrine Regulation in Aldosterone-producing Adenoma.

CONTEXT: Aldosterone-producing adenomas (APAs) are a common cause of primary aldosteronism (PA). Despite the discovery of somatic mutations in APA and the characterization of multiple factors regulating adrenal differentiation and function, the sequence of events leading to APA formation remains to be determined. OBJECTIVE: We investigated the role of Wnt/ß-catenin and adrenocorticotropin signaling, as well as elements of paracrine regulation of aldosterone biosynthesis in adrenals with APA and their relationship to intratumoral heterogeneity and mutational status. METHODS: We analyzed the expression of aldosterone-synthase (CYP11B2), CYP17A1, ß-catenin, melanocortin type 2 receptor (MC2R), phosphorlyated cAMP response element-binding protein (pCREB), tryptase, S100, CD34 by multiplex immunofluorescence, and immunohistochemistry-guided reverse transcription-quantitative polymerase chain reaction. Eleven adrenals with APA and 1 with micronodular hyperplasia from patients with PA were analyzed. Main outcome measures included localization of CYP11B2, CYP17A1, ß-catenin, MC2R, pCREB, tryptase, S100, CD34 in APA and aldosterone-producing cell clusters (APCCs). RESULTS: Immunofluorescence revealed abundant mast cells and a dense vascular network in APA, independent of mutational status. Within APA, mast cells were localized in areas expressing CYP11B2 and were rarely colocalized with nerve fibers, suggesting that their degranulation is not controlled by innervation. In these same areas, ß-catenin was activated, suggesting a zona glomerulosa cell identity. In heterogeneous APA with KCNJ5 mutations, MC2R and vascular endothelial growth factor A expression was higher in areas expressing CYP11B2. A similar pattern was observed in APCC, with high expression of CYP11B2, activated ß-catenin, and numerous mast cells. CONCLUSION: Our results suggest that aldosterone-producing structures in adrenals with APA share common molecular characteristics and cellular environment, despite different mutation status, suggesting common developmental mechanisms.

Efetividade de jogo educativo para gestantes: conhecimento agregado e vivência das mulheres / Eficacia del juego educativo para las mujeres embarazadas: conocimiento agregado y experiencia de las mujeres / Effectiveness of an educational game for pregnant women: aggregated knowledge and the women's experience

RESUMO Objetivos verificar a efetividade do jogo educativo para gestantes sobre seus direitos durante o trabalho de parto e parto; comparar o conhecimento das gestantes sobre seus direitos antes e após a aplicação do jogo e conhecer como foi para elas essa experiência. Método estudo quantitativo com análise qualitativa secundária. A coleta de dados ocorreu entre julho e setembro de 2019. Participaram 51 gestantes atendidas em duas Unidades Básicas de Saúde do município de Guarulhos. Utilizaram-se os testes Kolmogorov-Smirnov e o não-paramétrico Wilcoxon e nível de significância de 0,05. Adotou-se a análise lexical de conteúdo com apoio do software Interface de R pour les Analyses Multidimensionnelles de Textes et de Questionnaires. Resultados a efetividade do jogo foi comprovada pela comparação da média do conhecimento das gestantes antes e após a atividade educativa (Z = -5,924; p = 0,000). Houve diferença significativa na comparação da avaliação do conhecimento das gestantes antes e após o jogo (p=0,000). As respostas das gestantes enfatizaram a positividade da ação educativa e valorizaram as imagens do jogo como recurso educacional inovador. Conclusão e Implicações para a prática O jogo mostrou-se efetivo para ser utilizado na orientação de gestantes de forma lúdica e demonstrou agregar conhecimento de modo imagético e participativo.

RESUMEN Objetivos verificar la eficacia del juego educativo para mujeres embarazadas sobre sus derechos durante el trabajo de parto y el parto; comparar el conocimiento de las mujeres embarazadas sobre sus derechos antes y después de aplicar el juego y saber cómo fue para ellas esta experiencia. Método estudio cuantitativo con análisis cualitativo secundario. La recolección de datos se realizó entre julio y septiembre de 2019. Participaron 51 gestantes atendidas en dos Unidades de Atención Básicas de Salud de la ciudad de Guarulhos. Se utilizaron las pruebas de Kolmogorov-Smirnov y las pruebas no paramétricas de Wilcoxon. El nivel de significancia adoptado fue de 0,05. Se adoptó por el análisis de contenido léxico con el apoyo del Interface de R pour les Analyses Multidimensionnelles de Textes et de Questionnaires. Resultados el conocimiento medio de las embarazadas después de la actividad educativa fue mayor que el conocimiento medio antes de la actividad (Z = -5,924; p = 0,000). Hubo una diferencia significativa en la comparación de los conocimientos de las mujeres embarazadas antes y después del juego (p=0,000). Las respuestas de las embarazadas destacaron la positividad de la acción educativa y valoraron las imágenes del juego como un recurso educativo innovador. Conclusión e implicaciones para la práctica el juego demostró ser eficaz para ser utilizado en la orientación de las mujeres embarazadas de una manera lúdica y demostró añadir conocimiento de una manera imagética y participativa.

ABSTRACT Objectives to verify the effectiveness of an educational game for pregnant women about their rights during labor and delivery; to compare the knowledge of pregnant women about their rights before and after applying the game and to investigate how good this experience was for them. Method quantitative study with secondary qualitative analysis. Data collection took place between July and September 2019. Participants were 51 pregnant women attended at two Primary Health Units of the city of Guarulhos. The Kolmogorov-Smirnov and the non-parametric Wilcoxon tests were used. The level of significance adopted was .05. Lexical content analysis was performed with the use of the Interface de R pour les Analyses Multidimensionnelles de Textes et de Questionnaires software. Results the mean knowledge of the pregnant women after the educational activity was higher than the mean knowledge before the activity (Z = -5.924; p = .000). There was a significant difference when comparing the assessment of the pregnant women's knowledge before and after the game (p = .000). The responses of the pregnant women emphasized the positivity of the educational action and valued the images of the game as an innovative educational resource. Conclusion and Implications for practice The study demonstrated that the game was effective for use in guiding pregnant women playfully and that knowledge was gained in an imagetic and participatory way.