Assessing the gene silencing potential of AuNP-based approaches on conventional 2D cell culture versus 3D tumor spheroid

Publisher Copyright: Copyright © 2024 Oliveira, Fernandes and Baptista.Three-dimensional (3D) cell culture using tumor spheroids provides a crucial platform for replicating tissue microenvironments. However, effective gene modulation via nanoparticle-based transfection remains a challenge, often facing delivery hurdles. Gold nanoparticles (AuNPs) with their tailored synthesis and biocompatibility, have shown promising results in two-dimensional (2D) cultures, nevertheless, they still require a comprehensive evaluation before they can reach its full potential on 3D models. While 2D cultures offer simplicity and affordability, they lack physiological fidelity. In contrast, 3D spheroids better capture in vivo conditions, enabling the study of cell interactions and nutrient distribution. These models are essential for investigating cancer behavior, drug responses, and developmental processes. Nevertheless, transitioning from 2D to 3D models demands an understanding of altered internalization mechanisms and microenvironmental influences. This study assessed ASO-AuNP conjugates for silencing the c-MYC oncogene in 2D cultures and 3D tumor spheroids, revealing distinctions in gene silencing efficiency and highlighting the microenvironment’s impact on AuNP-mediated gene modulation. Herein, we demonstrate that increasing the number of AuNPs per cell by 2.6 times, when transitioning from a 2D cell model to a 3D spheroid, allows to attain similar silencing efficiencies. Such insights advance the development of targeted gene therapies within intricate tissue-like contexts.publishersversionpublishe

A política pública e as transferências voluntárias do turismo no Brasil : uma análise do período 2009- 2019

Trabalho de Conclusão de Curso (graduação)—Universidade de Brasília, Centro de Excelência em Turismo, 2020.As transferências voluntárias (TVs) da União se configuram em um processo importante para a redução do desequilíbrio fiscal entre as esferas públicas e para a equalização interregional, se constituindo em transferências de recursos públicos federais para ações executadas de forma descentralizada, que atendem demandas locais. O Ministério do Turismo (MTur), órgão público administrativo de máxima autoridade em sua esfera de competência, figura como o quarto órgão que mais disponibilizou recursos para ações de execução descentralizada, segundo o Painel de Transferências Abertas +Brasil, portal online de consulta sobre as Transferências da União realizadas pela Plataforma +Brasil. Assim, o trabalho aqui desenvolvido teve como objetivo descrever e analisar as transferências voluntárias do MTur de 2009 a 2019, com o intuito de se avaliar a ação discricionária da gestão pública ministerial e relacionar tais ações à política pública de turismo. A pesquisa foi desenvolvida orientada por metodologia exploratória e descritiva, de caráter qualitativo e quantitativo, a partir do levantamento de referenciais legais, bibliográficos e documentais, assim como investigação em portais online de acesso à informação para consultas e fonte de dados. Os resultados apurados demostram ineficiência técnica do MTur na gestão das TVs, disparidade regional na destinação do recurso e que, apesar de alinhadas aos diversos programas/linhas de ação dos Planos Nacionais de Turismo (PNTs), os investimentos em infraestrutura e eventos foram significativamente maiores.The voluntary transfers (TVs) of the Union, despite notorious criticisms, are configured in an important process for the reduction of the fiscal imbalance between the public spheres and for the interregional equalization, constituting in transfers of federal public resources for actions executed in a decentralized way, to meet local demands. The Ministry of Tourism (MTur), the administrative public body with maximum authority in its sphere of competence, was the fourth body that most provided resources for decentralized execution actions, according to the Painel de Transferências Abertas +Brasil, an online consultation portal on Transfers of the Union carried out by Plataforma + Brasil. Thus, the work developed here aimed to describe and analyze voluntary transfers from the MTur from 2009 to 2019, in order to evaluate the discretionary action of the ministerial public management and relate these actions to the public policy of tourism. The research was developed under exploratory and descriptive methodology, of qualitative and quantitative character, from the survey of legal, bibliographic and documentary references, as well as research in online portals of access to information for consultations and data source. The results obtained demonstrate the technical inefficiency of MTur in the management of TVs, regional disparity in the allocation of the resource and that, despite being aligned with various lines of action of the National Tourism Plans (PNTs), the investment in the infrastructure and events niches was significantly higher than in other actions

Children’s perceived barriers to a healthy diet: the influence of child and community-related factors

A healthy diet influences the promotion and maintenance of health throughout an individual’s life. Many individuals struggle to have a healthy diet, despite it being mainly under their control. The current study aims to explore children’s perceived barriers to a healthy diet. A qualitative study with the open-ended question, “Please identify the top 5 barriers to a healthy diet”, was undertaken between January–June 2019 in which 274 students from the 5–6th grades wrote down their answers to the open-ended question. Content analysis was used to analyze responses with a codebook based on the Six C’s Model. Five categories were identified: Child, Clan, Community, Country, and Culture-related barriers. Findings showed that the barriers most highlighted were in the Child sphere (e.g., dietary intake) and the Community sphere (e.g., peer food choices). Children seldom referred to barriers from the Clan sphere, i.e., related to family (e.g., food available at home). Additionally, it seems that girls emphasize more barriers from the Child sphere, while boys emphasize more barriers from the Community sphere. Due to the qualitative nature of this study, interpretation of the data should take into account the specific characteristics and context of the sample. Nevertheless, the current data are helpful in identifying implications for practice, for example, the need to empower children with tools (e.g., self-regulation-based interventions) likely to help them overcome perceived barriers. Finally, advocacy groups may help set environmental and structural changes in the community likely to facilitate children’s healthy choices.This study was conducted at the Psychology Research Centre (CIPsi/UM) School of Psychology, University of Minho, supported by the Foundation for Science and Technology (FCT) through the Portuguese State Budget (UIDB/01662/2020). Additionally, this study was supported by the Portuguese Foundation for Science and Technology and the Portuguese Ministry of Science, Technology and Higher Education through national funds (PTDC/PSI-GER/28302/2017) and cofinanced by FEDER through COMPETE2020 under the PT2020 Partnership Agreement (POCI-01- 0145-FEDER-028302). This study was also supported by the Portuguese Foundation for Science and Technology and the Portuguese Ministry of Science, Technology and Higher Education, through the national funds, within the scope of the Transitory Disposition of the Decree No. 57/2016, of 29th of August, amended by Law No. 57/2017 of 19 July. BP was supported by a Doctoral research grant (SFRH/BD/143469/2019) from the Portuguese Foundation for Science and Technology

Estudo da eficácia do uso da eletroneuromiografia como guia para aplicação de toxina botulínica

A distonia cervical é caracterizada por contrações musculares involuntárias, envolvendo as regiões da cabeça, pescoço e ombros. Trata-se da distonia focal mais comum, e sua prevalência é maior em mulheres com pico na quinta década de vida, mas pode afetar qualquer idade. O tratamento é essencialmente sintomático, a fim de aliviar as contrações musculares e dores. Atualmente o tratamento padrão ouro é a aplicação da toxina botulínica. A injeção da toxina é realizada a partir da avaliação anatômica. Contudo, devido a falta de precisão da técnica anatômica os pacientes podem apresentar alguns efeitos adversos como disfagia, fraqueza no pescoço, disartria, disfonia, dificuldade respiratória. A fim de aprimorar os resultados , foram desenvolvidas técnicas guiadas para essa aplicação como o uso da eletromiografia (EMG) ou ultrassonografia para melhorar a localização exata do músculo durante a aplicação da toxina, a fim de diminuir os efeitos adversos. Esse estudo teve como objetivo comparar o resultado da aplicação da toxina botulínica na distonia cervical por aplicação guiada por EMG versus a técnica da palpação anatômica. Os pacientes (n=7) foram submetidos escala Toronto Western Spasmodic Torticollis Rating Scale (TWSTRS), posteriormente receberam a aplicação guiada pelo EMG e foram reavaliados após duas semanas com a mesma escala e seus resultados subtraídos levando a diferença gerada pelo tratamento utilizado . Os resultados foram expostos por meio de tabela e gráficos e então comparados com os encontrados na literatura que utilizaram a mesma escala de avaliação. Foi concluído que a aplicação da toxina botulínica se mostrou como tratamento eficaz para melhorar a qualidade de vida desses pacientes, porém de forma muito semelhante quando por aplicação anatômica ou guiada com uso da EMG

Fast prototyping microfluidics: Integrating droplet digital lamp for absolute quantification of cancer biomarkers

UID/CTM/50025/2019 UID/Multi/04378/2019 Inn-INDIGO/0002/2015 PTDC/BTM-SAL/31201/2017 SFRH/BPD/124311/2016Microfluidic (MF) advancements have been leveraged toward the development of state-of-the-art platforms for molecular diagnostics, where isothermal amplification schemes allow for further simplification of DNA detection and quantification protocols. The MF integration with loop-mediated isothermal amplification (LAMP) is today the focus of a new generation of chip-based devices for molecular detection, aiming at fast and automated nucleic acid analysis. Here, we combined MF with droplet digital LAMP (ddLAMP) on an all-in-one device that allows for droplet generation, target amplification, and absolute quantification. This multilayer 3D chip was developed in less than 30 minutes by using a low-cost and extremely adaptable production process that exploits direct laser writing technology in “Shrinky-dinks” polystyrene sheets. ddLAMP and target quantification were performed directly on-chip, showing a high correlation between target concentration and positive droplet score. We validated this integrated chip via the amplification of targets ranging from five to 500,000 copies/reaction. Furthermore, on-chip amplification was performed in a 10 µL volume, attaining a limit of detection of five copies/µL under 60 min. This technology was applied to quantify a cancer biomarker, c-MYC, but it can be further extended to any other disease biomarker.publishersversionpublishe

Assistência de enfermagem à criança autista: revisão integrativa

Introduction: The Autistic Spectrum Disorder (ASD) configures a neurodevelopmental disorder that involves social interaction, communication and behavior identified generally in pre-school children. The nursing professionals must perform the management and monitoring of the autistic child.Objective: To examine the scientific evidence about nursing care to the autistic child.Methodology: An integrative review of the literature, held in databases: CINAHL, Web of Science and LILACS databases using the search terms: “Nursing Care”, “Child”, “Child, Preschool”, “Autistic Disorder” and “Autism Spectrum Disorder”. There were included articles published between the period of 2013 to 2017 in Portuguese, Spanish and English.Resulted: The articles included were presented in summary table and the analysis of the results was performed descriptively presenting the synthesis of studies through comparisons and highlight of differences and/or similarities. It was identified that is basic to nursing to have empathy, holistic vision and knowledge to perform singular assistance and of quality for the child and family.Conclusion: Nursing uses the empathy, holistic view and different strategies for the care to the autistic child; however, the professionals refer difficulties in clinical practice. The publications on the subject are scarce being necessary the development of clinical research.  Introducción: El Trastorno del Espectro Autista (TEA) configura un trastorno del neurodesarrollo que compromete la interacción social, comunicación y comportamiento, identificado generalmente, en el niño preescolar. Los profesionales de enfermería deben realizar el manejo y acompañamiento del niño autista.Objetivo: Analizar las evidencias científicas sobre la asistencia de Enfermería al niño autista.Metodología: Revisión integradora de la literatura realizada en la base de datos: CINAHL, Web of Science e LILACS, se utilizó como descriptores: Nursing Care/Cuidados de Enfermagem”, “Child/Criança”, “Child, Preschool/Pré-escolar”, “Autism Disorder/Transtorno Autístico” y “Autism Spectrum Disorder/Transtorno do Espectro Autista”. Se incluyeron artículos publicados entre el período de 2013 a 2017 en portugués, español e inglés.Resultados: Los artículos incluidos fueron presentados en cuadro sinóptico y el análisis de los resultados fue realizado de forma descriptiva presentando la síntesis de los estudios por medio de comparaciones y destaque de diferencias y / o semejanzas. Se identificó que es fundamental que la enfermería tenga empatía, visión holística y conocimiento para realizar asistencia singular y de calidad para el niño y la familia.Conclusión: La enfermería utiliza la empatía, visión holística y diferentes estrategias para el cuidado del niño autista, sin embargo, los profesionales refieren dificultades en la práctica clínica. Las publicaciones sobre la temática son escasas siendo necesario el desarrollo de investigaciones clínicas.Introdução: O Transtorno do Espectro Autista (TEA) configura uma perturbação do neurodesenvolvimento que compromete a interação social, comunicação e comportamento identificado geralmente na criança pré-escolar. Os profissionais de enfermagem devem realizar o manejo e acompanhamento da criança autista. Objetivo: Analisar as evidências científicas sobre a assistência de Enfermagem à criança autista.Metodologia: Revisão integrativa da literatura, realizada nas bases de dados: CINAHL, Web of Science e LILACS utilizando os termos de busca: “Nursing Care/Cuidados de Enfermagem”, “Child/Criança”, “Child, Preschool/Pré-escolar”, “Autism Disorder/Transtorno Autístico” e “Autism Spectrum Disorder/Transtorno do Espectro Autista”. Foram incluídos artigos publicados entre o período de 2013 a 2017 nos idiomas português, espanhol e inglês.Resultados: Os artigos incluídos foram apresentados em quadro sinóptico e a análise dos resultados foi realizada de forma descritiva apresentando a síntese dos estudos por meio de comparações e destaque de diferenças e/ou semelhanças. Identificou-se que é fundamental à enfermagem ter empatia, visão holística e conhecimento para realizar assistência singular e de qualidade para a criança e família. Conclusão: A enfermagem utiliza a empatia, visão holística e diferentes estratégias para o cuidado a criança autista, no entanto os profissionais referem dificuldades na pratica clínica. As publicações sobre a temática são escassas sendo necessário o desenvolvimento de pesquisas clínicas

Combining the amplification refractory mutation system and high-resolution melting analysis for KRAS mutation detection in clinical samples

© The Author(s) 2023. 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/.The success of personalized medicine depends on the discovery of biomarkers that allow oncologists to identify patients that will benefit from a particular targeted drug. Molecular tests are mostly performed using tumor samples, which may not be representative of the tumor's temporal and spatial heterogeneity. Liquid biopsies, and particularly the analysis of circulating tumor DNA, are emerging as an interesting means for diagnosis, prognosis, and predictive biomarker discovery. In this study, the amplification refractory mutation system (ARMS) coupled with high-resolution melting analysis (HRMA) was developed for detecting two of the most relevant KRAS mutations in codon 12. After optimization with commercial cancer cell lines, KRAS mutation screening was validated in tumor and plasma samples collected from patients with pancreatic ductal adenocarcinoma (PDAC), and the results were compared to those obtained by Sanger sequencing (SS) and droplet digital polymerase chain reaction (ddPCR). The developed ARMS-HRMA methodology stands out for its simplicity and reduced time to result when compared to both SS and ddPCR but showing high sensitivity and specificity for the detection of mutations in tumor and plasma samples. In fact, ARMS-HRMA scored 3 more mutations compared to SS (tumor samples T6, T7, and T12) and one more compared to ddPCR (tumor sample T7) in DNA extracted from tumors. For ctDNA from plasma samples, insufficient genetic material prevented the screening of all samples. Still, ARMS-HRMA allowed for scoring more mutations in comparison to SS and 1 more mutation in comparison to ddPCR (plasma sample P7). We propose that ARMS-HRMA might be used as a sensitive, specific, and simple method for the screening of low-level mutations in liquid biopsies, suitable for improving diagnosis and prognosis schemes.This work is financed by national funds from FCT—Fundação para a Ciência e a Tecnologia, I.P., in the scope of the project UIDP/04378/2020 and UIDB/04378/2020 of the Research Unit on Applied Molecular Biosciences—UCIBIO and the project LA/P/0140/2020 of the Associate Laboratory Institute for Health and Bioeconomy—i4HB. FCT-MCTES is also acknowledged for 2020.07660.BD for BBO. Open access funding provided by FCT|FCCN (b-on).info:eu-repo/semantics/publishedVersio

Implantação de serviço de reabilitação profissional: a experiência da UFMG

Visando implementar políticas de promoção da saúde no trabalho, a UFMG (Universidade Federal de Minas Gerais) criou em 1999 o Serviço de Atenção à Saúde do Trabalhador (SAST Pampulha/ UFMG). Entre outrasatividades, o SAST desenvolve um programa de reabilitação profissionalpara os servidores da instituição parcialmente incapacitados para o trabalho, tendo atendido 78 trabalhadores entre junho de 1999 e dezembro de 2001, dos quais 65 foram considerados aptos para o trabalho e 13 foram aposentados. Este estudo visa avaliar os resultados do programa e traçar o perfil dos atendidos, com base em questionários e protocolos aplicados a 51 servidores. Dentre os que retornaram ao trabalho, a maioria retomou a mesma função anterior no mesmo local, mas 7 1 % tiveram restrição de tarefas. Nos trabalhadores cujo resultado final da reabilitação foi a aposentadoria, o diagnóstico clínico mais comum foi doença do sistema osteomuscular e do tecido conjuntivo. Cerca de 34% deles exerciam serviços administrativos. Os resultados permitiram uma reflexão sobre o papel das várias figuras envolvidas no processo de reabilitação profissional do trabalhador parcialmente incapacitado. A possibilidade de discutir a reabilitação na perspectiva dos diferentes saberes técnico-científicos da equipe multiprofissional do SAST vem contribuindo para a definição de uma política de reabilitação profissional na UFMG.In order to implement policies to promote health in the workplace, the Federal University of the State of Minas Gerais (UFMG) created in 1999 the Worker Health Attention Service (SAST). Among other activities, SAST runs a professional rehabilitation program to assist partially disabled workers,having attended 78 workers between June 1999 and December 2001, ofwhich 65 were considered able to work and 13 were retired. This studyassesses the results of the program and dresses a profile of the workers,drawing on questionnaires and protocols applied to!51 of them. Amongthose who got back to work, most regained the same job at the same place as before, but 71 % faced some task restriction. For the workers who had to retire after rehabilitation, the most common clinical diagnosis was disorder of the osteomuscular system and of the conjunctive tissue; 34% of them performed administrative services. These results have allowed for fruitful thinking on the role of the various subjects involved in the rehabilitation of the partially disabled worker. The possibility of discussing the rehabilitation process from the point of view of the various approaches in the multiprofessional team involved in SAST has contributed to the development of the professional rehabilitation policy within the University

Saúde bucal de idosos: rastreio de lesões dos tecidos moles na prevenção do câncer bucal

Objective: To detect oral soft tissue injuries in older people. Method: A quantitative analytical cross-sectional study conducted in Fortaleza, Ceará. Individual interviews addressed age, gender, marital status, income, and education. The community oral health indicator was used to detect oral soft tissue injuries and their location. Results: Most of the 821 participants aged 60-100 years were women (580; 70.6%), attended school for up to 5 years (401; 48.8%), were illiterate (201; 24.5%), were retired (608; 74.1%), and received up to 2 wages (701; 85.4%). A total of 604 participants (73.6%) wore dentures. Injuries included red patches (152; 55.9%), blisters (58; 21.3%), lesions and/ or wounds (39; 14.3%), and white patches (30; 11%). Locations of injuries were the roof of the mouth (167; 61.4%), gums (62; 22.8%), cheeks (39; 14.3%), tongue (15; 5.5%), lips (15; 5.5%), and the floor of the mouth (12; 4.4%). Injuries were associated with age (p<0.001), retirement (p=0.005), education (p=0.010), dentures (p<0.001) and red patches (p<0.001). Conclusion: Tracking soft tissue injuries and referring older adults with suspected malignant lesions to the health team should be included as oral cancer identification and prevention measures. Furthermore, health care providers should raise older adults’ awareness of the importance of regular preventive examinations

Genetic polymorphisms of genes involved in oxidative stress and inflammatory management in oncopediatric patients with chemo-induced oral mucositis

Oral mucositis (OM) is a painful inflammatory oral condition that affects children who undergo chemotherapy. Oxidative stress is a known OM mediator and pro-inflammatory cytokines contribute to the amplification of the immune response. Objective: To investigate the possible associations of rs4880 (superoxide dismutase 2, SOD2 47 C/T), rs7943316 (catalase, CAT −21 A/T), rs1800629 (tumor necrosis factor α, TNF- α −308 G/A), and rs1800795 (interleukin 6, IL-6 −174 G/C) polymorphisms with chemo-induced OM occurrence and severity in oncopediatric patients. Methodology: We conducted a single-center, observational cross-sectional study with sample collection of oral epithelial cells from 95 children and adolescents with hematological cancers who underwent chemotherapy, followed by genomic DNA extraction. Single-nucleotide polymorphisms (SNPs) were assessed with PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism). Demographic data and information concerning OM occurrence were obtained from dental charts of the multidisciplinary oral care team. Information on OM severity was obtained from appropriately-filled Oral Assessment Guide records. Descriptive and inferential statistics were conducted with Student's T test, chi-squared test, and Fisher's exact test, with p≤0.05. Results: The mean age was 10 years-old and most patients were male individuals (57.89%). Female sex was considered a protective factor for OM occurrence (OR=4.83; CI=[1.14; 16.57]). The AA genotype for CAT was the most frequent amongst individuals with severe OM (p=0.04). The GA genotype for TNF- α was the most frequent amongst individuals without severe OM (p=0.03). For SOD2 and IL-6 , the most frequent genotypes were CT and GG respectively for all groups (p>0.05). Conclusion:The AA genotype for CAT −21 A/T was a tendency among the group with severe OM. Data on TNF- α −308 G/A were inconclusive. No associations were detected for SOD2 47 C/T and IL-6 −174 G/C polymorphisms in oncopediatric patients with chemo-induced oral mucositis