The role of GSH signaling in mitophagy induced by reactive oxygen species: potential target for neuroprotection in stroke?

Mitochondria are involved in numerous cellular processes, including energy production and cell death regulation. Simultaneously, these are the main cellular producers of reactive oxygen species (ROS), particularly when dysfunctional. ROS accumulation leads to oxidative stress which is associated with pathologies, such as stroke and neurodegenerative diseases. So, elimination of ROS-producing damaged mitochondria is key for maintaining cell homeostasis. Mitophagy, consists of the selective degradation of excessive or damaged mitochondria. The accumulation of ROS and mitochondrial depolarization stimulates PTEN-induced kinase 1 (PINK1)/PARKIN-mediated mitophagy. Concomitantly, ROS are counteracted by glutathione, a key antioxidant. ROS oxidize glutathione and stimulate protein S-glutathionylation, a post- translation modification capable of modulate protein function. Here, the main goal was to ex- plore the molecular mechanisms underlying mitophagy, in particular the role of ROS and/or glutathione signaling in mitophagy progression, with a special focus on protein S-glutathi- onylation. We hypothesized that mitophagy can be regulated by glutathione signaling, namely that the accumulation of mitochondrial ROS stimulates the production and oxidation of gluta- thione, which enhances/stimulates mitophagy by protein glutathionylation of mitophagy-re- lated proteins. Using human cell lines, SH-SY5Y and SH-SY5Y mitoQC, mitochondrial dysfunc- tion was stimulated with the classical uncoupler carbonyl cyanide m-chlorophenylhydrazone (CCCP). ROS generation and PINK1/PARKIN-dependent mitophagy were observed after CCCP treatment. Likewise, the catalytic subunit of the rate limiting enzyme for glutathione synthesis, Glutamate-Cysteine Ligase, is upregulated in CCCP-treated cells. Notably, we show for the first time that CCCP treatment induces protein S-glutathionylation of PINK1. Furthermore, we also found fluorescence measure on a microplate reader to be a possible new technique to assess mitophagy. Understating mitophagy mechanisms and how these are regulated is key to stimulate cytoprotection by preventing cell death. Thus, this can eventually contribute to new strategies against diseases characterized by mitochondrial damage, like stroke.As mitocôndrias são essenciais em processos celulares como produção de energia e regulação da morte celular. Simultaneamente, são as principais produtoras de espécies reativas de oxigênio (ROS), particularmente quando disfuncionais. A acumulação de ROS leva a stress oxidativo que está associado a patologias, como acidente vascular cerebral e doenças neuro- degenerativas. Assim, a eliminação de mitocôndrias danificadas produtoras de ROS é funda- mental para a homeostase celular. A mitofagia consiste na degradação seletiva de mitocôn- drias excessivas ou danificadas. Acumulação de ROS e despolarização mitocondrial estimulam mitofagia via quinase 1 induzida por PTEN (PINK1)/PARKIN. Concomitantemente, ROS são neutralizados pela glutationa, um antioxidante chave. As ROS oxidam a glutationa e estimulam s-glutationilização, uma modificação pós-traducional capaz de modular a função proteica. Aqui, o objetivo foi explorar os mecanismos subjacentes à mitofagia, particularmente o papel da sinalização de ROS e/ou glutationa na progressão da mitofagia, com foco especial na s- glutationilização. Colocamos a hipótese que a mitofagia pode ser regulada pela sinalização da glutationa, nomeadamente que a acumulação de ROS mitocondriais estimula a produção e oxidação de glutationa, que estimula a mitofagia por S-glutationilização de proteínas relacio- nadas com mitofagia. Em linhas celulares humanas, SH-SY5Y e SH-SY5Y mitoQC, disfunção mitocondrial foi estimulada com carbonyl cyanide m-chlorophenylhydrazone (CCCP). Geração de ROS e mitofagia dependente de PINK1/PARKIN foram observados após tratamento com CCCP. Igualmente, a subunidade catalítica da enzima limitante da síntese de glutationa, Gluta- mato-Cisteína Ligase, é sobre-expressa em células tratadas com CCCP. Notavelmente, mostra- mos que tratamento com CCCP induz glutationilização de PINK1. Além disso, também desco- brimos que medir fluorescência num leitor de microplacas é uma possível técnica para avaliar mitofagia. Compreender os mecanismos de mitofagia e como são regulados é fundamental para estimular a citoproteção prevenindo a morte celular. E eventualmente contribuir para novas estratégias contra doenças caracterizadas por danos mitocondriais, como o acidente vascular cerebral

Produção de conteúdos educativos em Astronomia - vantagens e desafios da aposta no digital

Este relatório corresponde a uma experiência de estágio final de mestrado que ocorreu entre 10 de agosto e 9 de novembro de 2020. Este período permitiu, quando a prioridade era a revisão de literatura, pesquisar como, ao longo dos anos, a comunidade científica abordou a possibilidade de utilizar plataformas digitais na produção e divulgação de conteúdos e conceitos científicos, e ao mesmo tempo confirmar como de facto a comunidade utilizou estes conceitos, quando em contexto de pandemia. Terminada a revisão de literatura, o estágio consistiu no desenho/preparação de atividades e participação em transmissões em direto pela internet (livestreams) do Instituto de Astrofísica e Ciências do Espaço (IA), para ter experiências em primeira mão na organização e moderação das mesmas. Isto permitiu dar ao estágio componentes práticas e confirmar os constrangimentos de preparar e adaptar uma atividade e objetivos às particularidades de o fazer a partir da alçada de uma universidade/ instituto de investigação, como um grupo de comunicação de ciência. Por fim, tive a oportunidade de desenhar uma atividade para o IA, procurando corresponder às expetativas e interesses do público de acordo com as possibilidades do grupo.This report corresponds to a final internship experience for this master’s degree, that took place during August 10th and November 9th, 2020. This period allowed, when the priority consisted on the revision of the literature, to research how, throughout the years, the scientific community researched and approached the possibility of using digital platforms in the production and dissemination of scientific contents and concepts, and at the same time, confirm how, in fact, the community used these concepts, when under the reality of a pandemic. When finished with the revision, the internship consisted in the design/preparation of activities and participation in livestreams at the Institute of Astrophysics and Space Sciences, to have first-hand experience on the organization and moderation of those events, so that I could have a practical experience of the difficulties of preparing and adapting an activity and goals to the fact that the event is organized under the umbrella of an university/ research institute, science communication group. Lastly, I had the opportunity to design an activity for the Institute, looking to correspond to expectation and interest of the audience with the possibilities of the group

Sex differences in functional connectivity between resting state brain networks in Autism Spectrum Disorder

© The Author(s) 2021. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.Functional brain connectivity (FBC) has previously been examined in autism spectrum disorder (ASD) between-resting-state networks (RSNs) using a highly sensitive and reproducible hypothesis-free approach. However, results have been inconsistent and sex differences have only recently been taken into consideration using this approach. We estimated main effects of diagnosis and sex and a diagnosis by sex interaction on between-RSNs FBC in 83 ASD (40 females/43 males) and 85 typically developing controls (TC; 43 females/42 males). We found increased connectivity between the default mode (DM) and (a) the executive control networks in ASD (vs. TC); (b) the cerebellum networks in males (vs. females); and (c) female-specific altered connectivity involving visual, language and basal ganglia (BG) networks in ASD-in suggestive compatibility with ASD cognitive and neuroscientific theories.VT received support from Fundação Ciência e Tecnologia (FCT) PhD fellowship (PD/BD/114460/2016) and paid by FCT DSAIPA/DS/0065/2018 Grant. DP was supported by the European Commission Seventh Framework Programme Marie Curie Career Integration Grant FP7-PEOPLE-2013-CIG-631952, the 2016 Bial Foundation Psychophysiology Grant Ref. 292/16, and the IF/00787/2014, LISBOA-01-0145-FEDER-030907 and DSAIPA/DS/0065/2018 FCT Grants, and the iMM Lisboa Director’s Fund Breakthrough Idea Grant 2016; and is co-founder and shareholder of the neuroimaging research services company NeuroPsyAI, Ltd. MA was supported by FCT Grant UID/MAT/00006/2013.info:eu-repo/semantics/publishedVersio

The role of optic nerve sheath ultrasonography in increased intracranial pressure: a systematic review and meta analysis

© 2023 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC license (http://creativecommons.org/licenses/by-nc/4.0/).Objectives: To review the optimal diagnostic cut-off of ultrasonographic optic nerve sheath diameter (ONSD) in the diagnosis of increased intracranial pressure (IICP). Methods: A systematic search was conducted of available studies assessing the use of ONSD ultrasonography in patients with suspected IICP. Meta-analysis of diagnostic accuracy of ultrasonographic ONSD was performed using a bivariate model of random effects to summarize pooled sensitivity and specificity. A summary receiver operating characteristics (SROC) curve was plotted. Accuracy measures associated with ONSD cut-off and predefined covariates were investigated with meta-regression. Results: We included 38 studies, comprising a total of 2824 patients. A total of 21 studies used invasive techniques as a reference standard estimation of IICP and meta-analysis revealed a pooled sensitivity of 0.90 (95% CI 0.85-0.93) and specificity of 0.87 (95% CI 0.80-0.91). Optimal ONSD cut-off values ranged between 4.1 mm and 7.2 mm. Meta-regression analysis showed that ONSD cut-off values of 5.6 to 6.3 mm were associated with higher pooled specificity compared to cut-off values of 4.9 to 5.5 mm (0.93, 95% CI 0.85-0.97 vs. 0.78, 95% CI 0.65-0.87; p = 0.036). Conclusions: Ultrasonography of ONSD shows a high diagnostic accuracy for IICP, with high pooled sensitivity and specificity. Additionally, larger cut-off values seem to significantly increase specificity without compromising sensitivity, which support their use as optimal ONSD cut-off. The overall high sensitivity of ultrasonographic ONSD suggests its usefulness as a screening tool for IIC, which may provide an estimate of when invasive methods are warranted. Clinical relevance: ONSD ultrasonography is a fast and cost-effective method with a high diagnostic accuracy to detect IICP. The optimum ONSD cut-off hasn't been established before, but we suggest the 5.6 to 6.3 mm range as the best for the diagnosis of IICP.DB has received funding from the Portuguese Foundation for Science and Technology [Fundação para a Ciência e Tecnologia – reference 2022.10667.BD] and Santa Casa da Misericórdia de Lisboa [Prémio João Lobo Antunes 2022].info:eu-repo/semantics/publishedVersio

A methanolic extract of Ganoderma lucidum fruiting body inhibits the growth of a gastric cancer cell line and affects cellular autophagy and cell cycle

Ganoderma lucidum is one of the most extensively studied mushrooms as a functional food and as a chemopreventive agent due to its recognized medicinal properties. Some G. lucidum extracts have shown promising antitumor potential. In this study, the bioactive properties of various extracts of G. lucidum, from both the fruiting body and the spores, were investigated. The most potent extract identified was the methanolic fruiting body extract, which inhibited the growth of a gastric cancer cell line (AGS) by interfering with cellular autophagy and cell cycle.Fundaçao para a Ciencia e a Tecnologia (FCT, Portugal) and COMPETE/QREN/EU for providing financial support to this work (research project PTDC/AGR-ALI/110062/2009), CIMO (strategic project PEst-OE/AGR/UI0690/2011), FCT for the grant of R.T.L. (SFRH/BPD/68787/2010) and QREN for the grants of F.S.R. and D.S. (NORTE-07-0124-FEDER-000023). The authors also thank Dr Gabriela Almeida for technical assistance in the screening assays and Prof. Anabela Martins from the Polytechnic Institute of Bragança for confrmation of the identification of G. lucidum samples. IPATIMUP, Associate Laboratory of the Portuguese Ministry of Science, Technology and Higher Education, is partially supported by FCT

Retinal neurodegeneration in different risk phenotypes of diabetic retinal disease

Diabetic retinopathy (DR) has been considered a microvascular disease, but it has become evident that neurodegeneration also plays a key role in this complex pathology. Indeed, this complexity is reflected in its progression which occurs at different rates in different type 2 diabetic (T2D) individuals. Based on this concept, our group has identified three DR progression phenotypes that might reflect the interindividual differences: phenotype A, characterized by low microaneurysm turnover (MAT <6), phenotype B, low MAT (<6) and increased central retinal thickness (CRT); and phenotype C, with high MAT (≥6). In this study, we evaluated the progression of DR neurodegeneration, considering ganglion cell+inner plexiform layers (GCL+IPL) thinning, in 170 T2D individuals followed for a period of 5 years, to explore associations with disease progression or risk phenotypes. Ophthalmological examinations were performed at baseline, first 6 months, and annually. GCL+IPL average thickness was evaluated by optical coherence tomography (OCT). Microaneurysm turnover (MAT) was evaluated using the RetMarkerDR. ETDRS level and severity progression were assessed in seven-field color fundus photography. In the overall population there was a significant loss in GCL+IPL (−0.147 μm/year), independently of glycated hemoglobin, age, sex, and duration of diabetes. Interestingly, this progressive thinning in GCL + IPL reached higher values in phenotypes B and C (−0.249 and −0.238 μm/year, respectively), whereas phenotype A remained relatively stable. The presence of neurodegeneration in all phenotypes suggests that it is the retinal vascular response to the early neurodegenerative changes that determines the course of the retinopathy in each individual. Therefore, classification of different DR phenotypes appears to offer relevant clarification of DR disease progression and an opportunity for improved management of each T2D individual with DR, thus playing a valuable role for the implementation of personalized medicine in DR.info:eu-repo/semantics/publishedVersio

Atitudes dos Estudantes de Enfermagem Perante a Pessoa com Doença Mental

Background: The stigmatizing attitudes of nursing students towards people with mental illness can influence their learning and the development of skills, compromising the provision of care as future nurses. Objectives: To know the attitudes of nursing students towards the person with mental illness and to verify the existing correlation between the attendance to the curricular units of Mental Health and Psychiatry and their attitudes towards the person with mental illness. Methodology: Quantitative, descriptive-correlational study. Convenience sample with 47 undergraduate nursing students, from a Higher Education Institution in the northern region of Portugal. Data collected through an online questionnaire, consisting of the Attribution Questionaire (AQ-27) (Sousa et al., 2008). Descriptive and inferential statistical analysis was used, using the Statistical Package for the Social Sciences (version 26). Results: The presence of moderate stigma was verified. There were statistically significant differences between the semester of the course on the categories Irritation [X2 (3) =14,416; P=0.002], Hazard [X2 (3) =11.650; P=0.009] and Fear [X2 (3) =12.523; P=0.006] and Pena [F (3.43) = 5.471; P=0.003]. Conclusion: Theoretical and practical teaching revealed a decrease in stigmatizing attitudes, as well as previous experience in Mental Health. Familiarity only influenced the Irritation category.Marco Contextual: Las actitudes estigmatizantes de los estudiantes de enfermería hacia las personas con enfermedad mental pueden influir en su aprendizaje y desarrollo de habilidades, comprometiendo la prestación de cuidados como futuros enfermeros. Objetivo: Objetivos: Conocer las actitudes de los estudiantes de enfermería hacia la persona con enfermedad mental y verificar la correlación existente entre la asistencia a las unidades curriculares de Salud Mental y Psiquiatría y sus actitudes hacia la persona con enfermedad mental. Metodología: Metodología: Estudio cuantitativo, descriptivo-correlacional. Muestra de conveniencia con 47 estudiantes de pregrado en enfermería de una Institución de Educación Superior en la región norte de Portugal. Datos recolectados a través de un cuestionario en línea, consistente en el Cuestionario de Atribución (AQ-27) (Sousa et al., 2008). Se utilizó análisis estadístico descriptivo e inferencial, utilizando el Paquete Estadístico para las Ciencias Sociales (versión 26). Resultados: Resultados: Se verificó la presencia de estigma moderado. Hubo diferencias estadísticamente significativas entre el semestre de la carrera en las categorías Irritación [X2 (3) =14.416; P=0,002], Riesgo [X2 (3) =11,650; P=0.009] y Miedo [X2(3)=12.523; P=0,006] y Peña [F (3,43) = 5,471; p=0,003]. Conclusión: La enseñanza teórica y práctica reveló una disminución de las actitudes estigmatizantes, así como la experiencia previa en Salud Mental. La familiaridad solo influyó en la categoría Irritación.Enquadramento: As atitudes estigmatizantes dos estudantes de enfermagem perante a pessoa com doença mental podem influenciar as suas aprendizagens e o desenvolvimento de competências, comprometendo a prestação de cuidados como futuros enfermeiros. Objetivos: Conhecer as atitudes dos estudantes de enfermagem perante a pessoa com doença mental e verificar a correlação existente entre a frequência às unidades curriculares de Saúde Mental e Psiquiatria e as suas atitudes perante a pessoa com doença mental. Metodologia: Estudo quantitativo, descritivo-correlacional. Amostra de conveniência com 47 estudantes do curso de licenciatura em enfermagem de uma Instituição de Ensino Superior da região norte de Portugal. Dados colhidos através de questionário on-line, constituído pelo Attribution Questionaire (AQ-27) (Sousa et al., 2008). Recorreu-se à análise estatística descritiva e inferencial, através do Statistical Package for the Social Sciences (versão 26). Resultados: Verificou-se a presença de estigma moderado. Observaram-se diferenças estatisticamente significativas entre o semestre do curso sobre as categorias Irritação [X2 (3) =14,416; P=0,002], Perigosidade [X2 (3) =11,650; P=0,009] e Medo [X2 (3) =12,523; P=0,006] e Pena [F (3,43) = 5,471; P=0,003]. Conclusão: O ensino teórico e prático revelou diminuição de atitudes estigmatizantes, bem como a experiência prévia em Saúde Mental. A familiaridade apenas influenciou a categoria Irritação

Leukocytes and Albumin in Canine Leishmaniasis

Background: Canine Leishmaniasis (CanL) is a multisystemic and chronic inflammatory disease characterized by nonspecific clinical manifestations. In CanL, inflammatory cells and chemical mediators released in response to the parasite play a role in disease development and progression. Alterations on hematological parameters have been documented in CanL. These changes can also be assessed in relation to systemic inflammation caused by this disease. The circulating leukocyte counting, such as neutrophils, as well as the albumin level, are considered direct indicators of an inflammatory host environment. Several studies point to the use of biomarkers on the assistance in diagnosis and prognosis of several canine pathologies. The present study investigated the Neutrophils to Lymphocyte Ratio (NLR), Albumin to Globulin Ratio (AGR), and Neutrophils to Albumin Ratio (NAR) on systemic inflammatory response induced by Canine Leishmaniasis (CanL).Materials, Methods &amp; Results: For this purpose, adult dogs with confirmed diagnosis to CanL were divided into symptomatic (SD, n = 33) and asymptomatic (AD, n = 20) dogs for L. infantum and control dogs (CD, n = 20). Routine hematological and biochemical parameters were determined in blood samples using a veterinary automatic hematology and biochemical analyzers. Asymptomatic dogs (AD) had a higher number of white blood cells and neutrophils (16.48 ± 4.93; 13.41 ± 3.60, respectively) in relation to symptomatic dogs (SD) (13.54 ± 5.13; 10.42± 3.69, respectively) (P = 0.015 and P &lt; 0.0001, respectively). Neutrophils to Lymphocyte Ratio (NLR) was higher in dogs with leishmaniasis (9.45 ± 3.76) than in healthy dogs (3.39 ± 1.19) (P &lt; 0.0001). Serum total proteins (STP) and globulins increased in CanL, while albumin and AGR decreased in CanL, when compared to CD and references values to canine species. Neutrophils to Albumin Ratio (NAR) was higher in AD and SD (5.02 ± 1.14; 4.79 ± 1.07, respectively) when compared to CD (2.36 ± 0.55) (P &lt; 0.0001). Discussion: As reported in scientific researches, dogs with Leishmaniasis present alterations in circulating cell counts. Based on these data, we decided to expand this information using the NLR as a parameter in an attempt to better clarify the changes in these cells in CanL. We observed that NLR was increased on CanL in relation to healthy dogs, which could be a consequence of relative neutrophilia rather than lymphopenia. Neutrophils to Lymphocyte Ratio (NLR) is a biomarker that conveys information about inflammatory conditions. An elevated NLR can reflect an upregulated innate immune response, since neutrophils are effector cells of innate immunity and are involved in several acute and chronic inflammatory processes. Albumin is an acute phase protein that is considered an immune-inflammatory biomarker, which can be found reduced systemically in progressive inflammatory response. Serum total proteins (STP) and globulins were increased in CanL. These data are already well documented in CanL, which serum globulins are mainly associated with the increase of acute phase proteins, cytokines, and increase of specific antibodies to Leishmaniainfantum. Our results showed neutrophilia with hypoalbuminemia in CanL. So, in an attempt to assess the relationship of these two available markers, we used NAR calculation in order to evaluate the changes induced by CanL. In this study NAR was higher in CanL when compared to control dogs. Thus, our data indicate that NLR and NAR could be used as biomarkers in veterinary medical clinics in order to assess inflammatory profile in CanL, mainly in asymptomatic dogs. These parameters obtained from routine blood tests might be useful as cost-effective, easily accessible and helpful markers in order to distinguish the inflammatory response intensity in CanL