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Emission fluorescence is one of the most versatile and powerful biophysical techniques used in several scientific subjects. It is extensively applied in the studies of proteins, their conformations, and intermolecular contacts, such as in protein-ligand and protein-protein interactions, allowing qualitative, quantitative, and structural data elucidation. This review, aimed to outline some of the most widely used fluorescence techniques in this area, illustrate their applications and display a few examples. At first, the data on the intrinsic fluorescence of proteins is disclosed, mainly on the tryptophan side chain. Predominantly, research to study protein conformational changes, protein interactions, and changes in intensities and shifts of the fluorescence emission maximums were discussed. Fluorescence anisotropy or fluorescence polarization is a measurement of the changing orientation of a molecule in space, concerning the time between the absorption and emission events. Absorption and emission indicate the spatial alignment of the molecule's dipoles relative to the electric vector of the electromagnetic wave of excitation and emitted light, respectively. In other words, if the fluorophore population is excited with vertically polarized light, the emitted light will retain some polarization based on how fast it rotates in solution. Therefore, fluorescence anisotropy can be successfully used in protein-protein interaction investigations. Then, green fluorescent proteins (GFPs), photo-transformable fluorescent proteins (FPs) such as photoswitchable and photoconvertible FPs, and those with Large Stokes Shift (LSS) are disclosed in more detail. FPs are potent tools for the study of biological systems. Their versatility and wide range of colours and properties allow many applications. Finally, the application of fluorescence in life sciences is exposed, especially the application of FPs in fluorescence microscopy techniques with super-resolution that enables precise in vivo photolabeling to monitor the movement and interactions of target proteins.

Abstract Mutations in the EYS gene are one of the major causes of autosomal recessive retinitis pigmentosa. EYS-retinopathy presents a severe clinical phenotype, and patients currently have no therapeutic options. The progress in personalised medicine and gene and cell therapies hold promise for treating this degenerative disease. However, lack of understanding and incomplete comprehension of disease's mechanism and the role of EYS in the healthy retina are critical limitations for the translation of current technical advances into real therapeutic possibilities. This review recapitulates the present knowledge about EYS-retinopathies, their clinical presentations and proposed genotype–phenotype correlations. Molecular details of the gene and the protein, mainly based on animal model data, are analysed. The proposed cellular localisation and roles of this large multi-domain protein are detailed. Future therapeutic approaches for EYS-retinopathies are discussed.

In a highly competitive and liberalized energy market, where the retail of electricity is open to many potential companies, it is essential to have tools that help make decisions and guide the design of marketing strategies. In this sense, it is essential for retailers to know the behavior of their customers to correctly define their commercial strategies. One of the most commonly used methods for this is the characterization of their consumption profiles. Fortunately, for regulatory reasons, in some countries, the monthly electricity demand of each customer is openly available to any competitor. This paper explores whether this information, especially the economic sector and geographic location of a client, is useful for determining the client’s demand profile. Specifically, data on electricity demand in Spain from more than 27 million users and for a period of 3 years are analyzed. For this purpose, the electricity consumption of every client is grouped by month and normalized. The resulting demand profiles are later clustered according to different criteria. The main finding of the research is that the combined information on economic activity and location definitely enables prediction of the demand profile. Additionally, profile quality metrics are defined and obtained for the entire dataset. The resulting profiles have a mean dispersion of 10% and a confidence interval of ±17%. To clarify the use of these metrics, several examples are detailed, showing how this profile information can be used to improve the marketing decision-making process for electricity retailers.

A Teoria Antropológica do Didático considera dois aspectos da atividade humana: o estrutural, descrito em termos de praxeologias, e o funcional, analisado por meio da teoria dos Momentos de Estudo. Essa teoria tem sido utilizada em diversas pesquisas, inclusive na área do Ensino de Ciências, e tem evidenciado especificidades e desafios a docentes, destacando a necessidade de formação contínua destes. Nesse sentido, o objetivo deste artigo foi buscar aproximações entre a Teoria Antropológica do Didático e uma formação docente para o Ensino por Investigação, uma abordagem didática construtivista que tem o problema como ponto de partida. A presente pesquisa é qualitativa incluindo uma análise bibliográfica. A formação analisada foi desenvolvida pela pesquisa de Mestrado de Moura (2016) e, mais especificamente, duas atividades incluídas nessa formação, bem como a aplicação em sala de aula por algumas cursistas. Dentre os Momentos de Estudo identificados destacamos a institucionalização e a avaliação como premissas importantes a serem consideradas em formações. A análise em questão indicou o assujeitamento das professoras cursistas às prerrogativas do Ensino por Investigação após o curso. Acreditamos que a Teoria Antropológica do Didático é uma ferramenta teórica e analítica potente, sendo os Momentos de Estudo promissores para realização de futuras pesquisas sobre Ensino por Investigação e formação docente.

Retinitis pigmentosa (RP) is an inherited retinal degenerative disease. Mutations in EYS have been associated with autosomal recessive RP. The human iPS cell line, CABi002-A, derived from peripheral blood mononuclear cells from a patient carrying a heterozygous double mutation in EYS gene was generated by non-integrative reprogramming technology, using hOCT3/4, hSOX2, hc-MYC and hKLF4 reprogramming factors. Pluripotency and differentiation capacity were assessed by immunocytochemistry and RT-PCR. This iPSC line can be further differentiated towards the affected cells to understand the pathophysiology of the disease and test new therapeutic strategies.

Retinal dystrophies associated to mutations in the CRB1 gene comprise a wide array of clinical presentations. A blood sample from a patient with a family history of CRB1-retinal dystrophy was used to prepare the iPSC line ESi082-A. The genotype of the donor, affected of a perifoveal-bilateral macular dystrophy includes one frameshift deletion and one hypomorphic allele. ESi082-A cell line has been characterized for pluripotency and will be used to prepare retinal cellular models to study the dysfunction leading to the disease.

Wastewater-based epidemiology (WBE) has proven to be a powerful tool for the population-level monitoring of pathogens, particularly severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). For assessment, several wastewater sampling regimes and methods of viral concentration have been investigated, mainly targeting SARS-CoV-2. However, the use of passive samplers in near-source environments for a range of viruses in wastewater is still under-investigated. To address this, near-source passive samples were taken at four locations targeting student hall of residence. These were chosen as an exemplar due to their high population density and perceived risk of disease transmission. Viruses investigated were SARS-CoV-2 and its variants of concern (VOCs), influenza viruses, and enteroviruses. Sampling was conducted either in the morning, where passive samplers were in place overnight (17 h) and during the day, with exposure of 7 h. We demonstrated the usefulness of near-source passive sampling for the detection of VOCs using quantitative polymerase chain reaction (qPCR) and next-generation sequencing (NGS). Furthermore, several outbreaks of influenza A and sporadic outbreaks of enteroviruses (some associated with enterovirus D68 and coxsackieviruses) were identified among the resident student population, providing evidence of the usefulness of near-source, in-sewer sampling for monitoring the health of high population density communities. [ABSTRACT FROM AUTHOR]