Quantifying differentiation of progenitor populations using cerebral organoid models for neurodevelopmental disorders.

Neurodevelopmental disorders are characterized by complex phenotypes that often result from concomitant dysregulation of cell proliferation, differentiation, or other crucial developmental processes. Here, we present a protocol to quantify differentiation of progenitor populations during early stages of neurogenesis in induced pluripotent stem cell (iPSC)-derived cerebral organoids. We describe steps for organoid differentiation and maturation, sample preparation, immunofluorescence, and imaging and analysis using epifluorescence microscopy. This protocol can be used to compare cerebral organoids from control and patient-derived iPSCs. For complete details on the use and execution of this protocol, please refer to Rakotomamonjy et al. (2023).1.

Tecnologias educacionais utilizadas para promoção do autocuidado de pessoas com diabetes mellitus: revisão integrativa.

OBJECTIVE: To summarize the scientific productions that used educational technologies to promote self-care for people with diabetes. METHOD: Integrative review carried out from October 2022 to January 2023, in the databases: LILACS; Scopus; Embase; PubMed/MEDLINE and CINAHL. The search was paired, and the sample consisted of ten articles. The levels of evidence were analyzed by the Hierarchy of Evidence for Intervention Studies, and the results were synthesized for interpretation of the findings. RESULTS: The educational technologies identified to promote self-care were: mobile applications, interactive platforms, print, telemonitoring, video and simulation. The focus of the contents was on the promotion of foot care, prevention of neuropathy, self-management, knowledge, and expectation of people with diabetes and prevention of acute complications. CONCLUSION: The synthesis of knowledge about educational technologies to promote self-care for people with diabetes pointed to the need for robust evidence.

PCDH12 loss results in premature neuronal differentiation and impeded migration in a cortical organoid model.

Protocadherins (PCDHs) are cell adhesion molecules that regulate many essential neurodevelopmental processes related to neuronal maturation, dendritic arbor formation, axon pathfinding, and synaptic plasticity. Biallelic loss-of-function variants in PCDH12 are associated with several neurodevelopmental disorders (NDDs). Despite the highly deleterious outcome resulting from loss of PCDH12, little is known about its role during brain development and disease. Here, we show that PCDH12 loss severely impairs cerebral organoid development, with reduced proliferative areas and disrupted laminar organization. 2D models further show that neural progenitor cells lacking PCDH12 prematurely exit the cell cycle and differentiate earlier when compared with wild type. Furthermore, we show that PCDH12 regulates neuronal migration and suggest that this could be through a mechanism requiring ADAM10-mediated ectodomain shedding and/or membrane recruitment of cytoskeleton regulators. Our results demonstrate a critical involvement of PCDH12 in cortical organoid development, suggesting a potential cause for the pathogenic mechanisms underlying PCDH12-related NDDs.

Tecnologias educacionais utilizadas para promoção do autocuidado de pessoas com diabetes mellitus: revisão integrativa / Tecnologías educacionales utilizadas para la promoción del autocuidado de las personas con diabetes mellitus: Revisión Integradora / Tecnologias educacionais utilizadas para promoção do autocuidado de pessoas com diabetes mellitus: revisão integrativa

ABSTRACT Objective: To summarize the scientific productions that used educational technologies to promote self-care for people with diabetes. Method: Integrative review carried out from October 2022 to January 2023, in the databases: LILACS; Scopus; Embase; PubMed/MEDLINE and CINAHL. The search was paired, and the sample consisted of ten articles. The levels of evidence were analyzed by the Hierarchy of Evidence for Intervention Studies, and the results were synthesized for interpretation of the findings. Results: The educational technologies identified to promote self-care were: mobile applications, interactive platforms, print, telemonitoring, video and simulation. The focus of the contents was on the promotion of foot care, prevention of neuropathy, self-management, knowledge, and expectation of people with diabetes and prevention of acute complications. Conclusion: The synthesis of knowledge about educational technologies to promote self-care for people with diabetes pointed to the need for robust evidence.

RESUMEN Objetivo: Sintetizar las producciones científicas que utilizaron tecnologías educacionales para promoción del autocuidado de personas con diabetes. Método: Revisión integradora realizada entre octubre de 2022 hasta enero de 2023, en las bases de datos: LILACS; Scopus; Embase; PubMed/MEDLINE y CINAHL. La búsqueda ocurrió de manera pareada, y la muestra fue construida por diez artículos. Los niveles de evidencia fueron analizados por el Hierarchy of Evidence for Intervention Studies, y los resultados fueron sintetizados para interpretación de los hallados. Resultados: Las tecnologías educacionales identificadas para promoción del autocuidado fueron: aplicativos móviles, plataformas interactivas, impresos, telemonitorización, vídeo y simulado. El enfoque de los contenidos fue en la promoción del cuidado con los pies, prevención de neuropatia, autogestión, conocimiento y expectativa de personas con diabetes y prevención de complicaciones agudas. Conclusión: El síntesis del conocimiento sobre las tecnologías educacionales para promoción del autocuidado de personas con diabetes señaló la necesidad de evidencias robustas.

RESUMO Objetivo: Sumarizar as produções científicas que utilizaram tecnologias educacionais para promoção do autocuidado de pessoas com diabetes. Método: Revisão integrativa realizada no período de outubro de 2022 a janeiro de 2023, nas bases de dados: LILACS; Scopus; Embase; PubMed/MEDLINE e CINAHL. A busca aconteceu de forma pareada, e a amostra foi constituída por dez artigos. Os níveis de evidência foram analisados pelo Hierarchy of Evidence for Intervention Studies, e os resultados foram sintetizados para interpretação dos achados. Resultados: As tecnologias educacionais identificadas para promoção do autocuidado foram: aplicativos móveis, plataformas interativas, impressos, telemonitoramento, vídeo e simulação. O enfoque dos conteúdos foi na promoção do cuidado com os pés, prevenção de neuropatia, autogestão, conhecimento e expectativa de pessoas com diabetes e prevenção de complicações agudas. Conclusão: A síntese do conhecimento sobre as tecnologias educacionais para promoção do autocuidado de pessoas com diabetes apontou a necessidade de evidências robustas.

Cell-Laden Gradient Hydrogel Scaffolds for Neovascularization of Engineered Tissues.

Gradients in mechanical properties, physical architecture and biochemical composition exist in a variety of complex tissues, yet 3D in vitro models that enable investigation of these cues on cellular processes, especially those contributing to vascularization of engineered tissues are limited. Here, a photopolymerization approach to create cell-laden hydrogel biomaterials with decoupled and combined gradients in modulus, immobilized cell adhesive peptide (RGD) concentration, and proteolytic degradation enabling spatial encapsulation of vascular spheroids is reported to elucidate their impact on vascular sprouting in 3D culture. Vascular spheroids encapsulated in these gradient scaffolds exhibit spatial variations in total sprout length. Scaffolds presenting an immobilized RGD gradient promote biased vascular sprouting toward increasing RGD concentration. Importantly, biased sprouting is found to be dependent on immobilized RGD gradient characteristics, including magnitude and slope, with increases in these factors contributing to significant enhancements in biased sprouting responses. Conversely, reduction in biased sprouting responses is observed in combined gradient scaffolds possessing opposing gradients in RGD and modulus. The presented work is the first to demonstrate the use of a cell-laden biomaterial platform to systematically investigate the role of multiple scaffold gradients as well as gradient slope, magnitude and orientation on vascular sprouting responses in 3D culture.

Immobilized RGD concentration and proteolytic degradation synergistically enhance vascular sprouting within hydrogel scaffolds of varying modulus.

Insufficient vascularization limits the volume and complexity of engineered tissue. The formation of new blood vessels (neovascularization) is regulated by a complex interplay of cellular interactions with biochemical and biophysical signals provided by the extracellular matrix (ECM) necessitating the development of biomaterial approaches that enable systematic modulation in matrix properties. To address this need poly(ethylene) glycol-based hydrogel scaffolds were engineered with a range of decoupled and combined variations in integrin-binding peptide (RGD) ligand concentration, elastic modulus and proteolytic degradation rate using free-radical polymerization chemistry. The modularity of this system enabled a full factorial experimental design to simultaneously investigate the individual and interaction effects of these matrix cues on vascular sprout formation in 3 D culture. Enhancements in scaffold proteolytic degradation rate promoted significant increases in vascular sprout length and junction number while increases in modulus significantly and negatively impacted vascular sprouting. We also observed that individual variations in immobilized RGD concentration did not significantly impact 3 D vascular sprouting. Our findings revealed a previously unidentified and optimized combination whereby increases in both immobilized RGD concentration and proteolytic degradation rate resulted in significant and synergistic enhancements in 3 D vascular spouting. The above-mentioned findings would have been challenging to uncover using one-factor-at-time experimental analyses.

Thermal Analysis of a Magnetic Brake Using Infrared Techniques and 3D Cell Method with a New Convective Constitutive Matrix.

In this work we analyse the temperature distribution in a conductor disk in transitory regime. The disk is in motion in a stationary magnetic field generated by a permanent magnet and so, the electric currents induced inside it generate heat. The system acts as a magnetic brake and is analysed using infrared sensor techniques. In addition, for the simulation and analysis of the magnetic brake, a new thermal convective matrix for the 3D Cell Method (CM) is proposed. The results of the simulation have been verified by comparing the numerical results with those obtained by the Finite Element Method (FEM) and with experimental data obtained by infrared technology. The difference between the experimental results obtained by infrared sensors and those obtained in the simulations is less than 0.0459%.

Web building and silk properties functionally covary among species of wolf spider.

Although phylogenetic studies have shown covariation between the properties of spider major ampullate (MA) silk and web building, both spider webs and silks are highly plastic so we cannot be sure whether these traits functionally covary or just vary across environments that the spiders occupy. As MaSp2-like proteins provide MA silk with greater extensibility, their presence is considered necessary for spider webs to effectively capture prey. Wolf spiders (Lycosidae) are predominantly non-web building, but a select few species build webs. We accordingly collected MA silk from two web-building and six non-web-building species found in semirural ecosystems in Uruguay to test whether the presence of MaSp2-like proteins (indicated by amino acid composition, silk mechanical properties and silk nanostructures) was associated with web building across the group. The web-building and non-web-building species were from disparate subfamilies so we estimated a genetic phylogeny to perform appropriate comparisons. For all of the properties measured, we found differences between web-building and non-web-building species. A phylogenetic regression model confirmed that web building and not phylogenetic inertia influences silk properties. Our study definitively showed an ecological influence over spider silk properties. We expect that the presence of the MaSp2-like proteins and the subsequent nanostructures improves the mechanical performance of silks within the webs. Our study furthers our understanding of spider web and silk co-evolution and the ecological implications of spider silk properties.

Nanostructural and mechanical property changes to spider silk as a consequence of insecticide exposure.

Neonicotinoids are one of the world's most extensively used insecticides, but their sub-lethal influences on non-target and beneficial organisms are not well known. Here we exposed the orb web spider Parawixia audax, which is found on arable lands in Uruguay, to a sub-lethal concentration of the broad spectrum insecticide Geonex (thiamethoxam + lambda-cyhalothrin) and monitored their web building. We collected their major ampullate silk and subjected it to tensile tests, wide-angle X-ray diffraction (WAXS) analysis, and amino acid composition analysis. Around half of the exposed spiders failed to build webs. Those that built webs produced irregular webs lacking spiral threads. The mechanical properties, nanostructures, and amino acid compositions of the silk were all significantly affected when the spiders were exposed to insecticides. We found that silk proline, glutamine, alanine and glycine compositions differed between treatments, indicating that insecticide exposure induced downregulation of the silk protein MaSp2. The spiders in the control group had stronger, tougher and more extensible silks than those in the insecticide exposed group. Our WAXS analyses showed the amorphous region nanostructures became misaligned in insecticide exposed silks, explaining their greater stiffness. While the insecticide dose we subjected P. audax to was evidently sub-lethal, the changes in silk physicochemical properties and the impairment to web building will indelibly affect their ability to catch prey.

Técnicas invasivas y no invasivas en el diagnóstico de embriones humanos / Invasive and non-invasive techniques in human embryos diagnostic

Las técnicas de reproducción asistida se han desarrollado con el objetivo de aumentar las tasas de implantación y reducir el número de embriones transferidos. El conocimiento sobre la morfología embrionaria y el desarrollo de la tecnología molecular aplicada al diagnóstico preimplantacional, han supuesto un gran avance en el rendimiento de los ciclos, mejorando los resultados clínicos y disminuyendo la tasa de aborto. El objetivo de esta revisión es describir el estado actual de las técnicas de diagnóstico embrionario. Técnicas como el time-lapse aplicada a la morfocinética nos permiten estudiar de manera continua el desarrollo embrionario, aportando un enfoque más dinámico. Tecnología molecular como la metabolómica dirigida o la proteómica, muestran nuevos elementos de evaluación sobre las primeras etapas del desarrollo hasta blastocisto. Estudios sobre el secretoma embrionario en medios de cultivo in-vitro intentan predecir patrones de desarrollo en los embriones. La búsqueda de estrategias de diagnóstico y biomarcadores asociados a calidad embrionaria, aneuploidías y potencial implantatorio son nuevos retos a los que se enfrenta la tecnología en medicina reproductiva

The assisted reproduction techniques have been developed with the aim of increasing implantation rates and reducing the number of embryos transferred. Knowledge about embryonic morphology and the development of molecular technology applied to preimplantation diagnosis has resulted in great progress in terms of cycle performance, improving clinical outcomes and reducing the rate of abortion. The aim of this review is to describe embryonic diagnostic techniques. Currently, techniques like time-lapse applied to morphokinetics allow us to continuously study embryonic development providing a more dynamic approach. Molecular techniques such as directed metabolomics or proteomics show new elements of evaluation since the early stages of development to blastocyst. Studies on the metabolism of embryos cultured in-vitro are providing data that improve embryo selection. The development of diagnostic strategies and biomarkers associated with embryonic quality, aneuploidy and implantatory potential are new challenges for the future in reproductive medicine technology

Electro-quasistatic analysis of an electrostatic induction micromotor using the cell method.

An electro-quasistatic analysis of an induction micromotor has been realized by using the Cell Method. We employed the direct Finite Formulation (FF) of the electromagnetic laws, hence, avoiding a further discretization. The Cell Method (CM) is used for solving the field equations at the entire domain (2D space) of the micromotor. We have reformulated the field laws in a direct FF and analyzed physical quantities to make explicit the relationship between magnitudes and laws. We applied a primal-dual barycentric discretization of the 2D space. The electric potential has been calculated on each node of the primal mesh using CM. For verification purpose, an analytical electric potential equation is introduced as reference. In frequency domain, results demonstrate the error in calculating potential quantity is neglected (<3‰). In time domain, the potential value in transient state tends to the steady state value.

Para el análisis tiene usted que venir en ayunas / You must fast for the analysis

Objetivos. Estudiar la cumplimentación del ayuno correcto previo a la realización del análisis de sangre y averiguar si los profesionales sanitarios aconsejan el ayuno de 12 horas. Sujetos y métodos: Se pasó una encuesta a todos los adultos que se realizaron una analítica de sangre en un centro de salud rural (CSR) y en un laboratorio hospitalario (LH) durante periodos de dos y una semanas respectivamente. Se recogieron los datos de: sexo, edad, si comieron después de las 20 horas del día previo, si sabían que debían guardar ayuno y si algún profesional sanitario les advirtió de mantener un ayuno de 12 horas. Resultados: En el CSR se recogieron 202 encuestas, el 58,1% eran mujeres y la edad media de 53,7 ± 18,7. Conocían la importancia del ayuno el 95,3% y comieron después de las 20 horas del día previo el 66,5%. Al 86,6% nadie les dijo que debían guardar ayuno de 12 horas. En el LH se recogieron 243 encuestas, el 54% eran hombres y la edad media, de 54,1 ± 18,7. Conocían la importancia del ayuno el 96,5% y comieron después de las 20 horas el 78,2%. Al 90,2% nadie les dijo que debían guardar ayuno de 12 horas. Conclusiones: La mayoría de la población conoce la importancia del ayuno. El porcentaje de observancia del ayuno de 12 horas es muy bajo, aunque algo mayor en la población rural. Los sanitarios no informamos adecuadamente a nuestros pacientes (AU)

Objetives: 1. Study correct fasting compliance prior to the performance of the blood analysis. 2. Discover if the health care professionals have recommended 12-hour fasting. Subjects and methods: All the adults who underwent a blood test in a rural health care center (RHCS) and in a hospital laboratory (HL) during a two week and one week period, respectively, were surveyed. The survey collected: gender, age, if the subject had eaten after 8 p.m. on the day before the test, if they had known they had to fast and if any health care professional had told them to maintain a 12-hour fast. Results: In the RHCS, 202 surveys were collected (58.1% women). Mean age was 53.7 ± 18.7. A total of 95.3% had known the importance of fasting, and 66.5% had eaten after 8 p.m. the day before the test. A total of 86.6% had not been informed they had to fast for 12 hours. In the HL, 243 surveys were collected, 54% men. Mean age 54.1 ± 18.7. A total of 96.5% knew about the importance of fasting, and 78.2 had eaten after 8 p.m. the day before. A total of 90.2% had not been informed of the need to follow a 12- fasting period. Conclusions: Most of the population knows the importance of fasting. However, observance of a 12-hour fasting period is low, somewhat greater in the rural population. The health care professionals do not adequately inform their patients (AU)