Light-sheet microscopy: a tutorial

This paper is intended to give a comprehensive review of light-sheet (LS) microscopy from an optics perspective. As such, emphasis is placed on the advantages that LS microscope configurations present, given the degree of freedom gained by uncoupling the excitation and detection arms. The new imaging properties are first highlighted in terms of optical parameters and how these have enabled several biomedical applications. Then, the basics are presented for understanding how a LS microscope works. This is followed by a presentation of a tutorial for LS microscope designs, each working at different resolutions and for different applications. Then, based on a numerical Fourier analysis and given the multiple possibilities for generating the LS in the microscope (using Gaussian, Bessel, and Airy beams in the linear and nonlinear regimes), a systematic comparison of their optical performance is presented. Finally, based on advances in optics and photonics, the novel optical implementations possible in a LS microscope are highlighted.Peer ReviewedPostprint (published version

Conspiracy theories, credibility and trust in information

The rapid spread of social media on the Internet has resulted in strong changes in the information and communication landscape. Their inadequate use has given rise to new discussions on truth and post-truth, what is trustworthy and what is questionable, the credibility of messages and their sources. Through this article we aim to examine what citizens believe about the information they receive and if they feel whether or not there is withholding of information. This will be achieved through data obtained from a survey of a statistically representative sample of the Andalusian population over the age of 18 (1,103 respondents, with a maximum margin of error of +/- 3%). One of the main results of this research is the serious lack of credibility of the information received, as many Andalusians (68.1%) believe that information is withheld from them. We have also observed several factors explaining a greater likelihood of finding a high belief in the conspiracy theory on the withholding of information. Finally, the article reflects on the consequences of mistrust in information and we propose an inter- and transdisciplinary approach in order to counteract this mistrust.La rápida extensión del uso de redes sociales en internet ha provocado intensos cambios en el panorama informativo y de la comunicación. Un uso inadecuado de las mismas ha originado nuevos debates sobre la verdad y posverdad, lo confiable y lo dudoso, la credibilidad de los mensajes y de sus fuentes. A través de este artículo, basado en la realización de una encuesta realizada a una muestra estadísticamente representativa a la población andaluza mayor de 18 años (1.103 entrevistados, con un margen de error máximo de +/- 3 %), queremos examinar qué piensan los ciudadanos respecto a la información que reciben y si aprecian que hay ocultación de información o no. Uno de los principales resultados de esta investigación es la elevada falta de credibilidad de la información recibida, al constatarse la alta creencia de los andaluces en que se les oculta información (68,1 %). También hemos observado diversos factores que explican una mayor probabilidad de encontrar una alta creencia en la teoría de la conspiración sobre la ocultación de información. Termina el artículo reflexionando sobre las consecuencias de la falta de confianza en la información y se propone un enfoque inter y transdisciplinar para contrarrestarla

CRISPR-mediated reactivation of DKK3 expression attenuates TGF-beta signaling in prostate cancer

The DKK3 gene encodes a secreted protein, Dkk-3, that inhibits prostate tumor growth and metastasis. DKK3 is downregulated by promoter methylation in many types of cancer, including prostate cancer. Gene silencing studies have shown that Dkk-3 maintains normal prostate epithelial cell homeostasis by limiting TGF-β/Smad signaling. While ectopic expression of Dkk-3 leads to prostate cancer cell apoptosis, it is unclear if Dkk-3 has a physiological role in cancer cells. Here, we show that treatment of PC3 prostate cancer cells with the DNA methyltransferase (DNMT) inhibitor decitabine demethylates the DKK3 promoter, induces DKK3 expression, and inhibits TGF-β/Smad-dependent transcriptional activity. Direct induction of DKK3 expression using CRISPR-dCas9-VPR also inhibited TGF-β/Smad-dependent transcription and attenuated PC3 cell migration and proliferation. These effects were not observed in C4-2B cells, which do not respond to TGF-β. TGF-β signals can regulate gene expression directly via SMAD proteins and indirectly by increasing DNMT expression, leading to promoter methylation. Analysis of genes downregulated by promoter methylation and predicted to be regulated by TGF-β found that DKK3 induction increased expression of PTGS2, which encodes cyclooxygenase-2. Together, these observations provide support for using CRISPR-mediated induction of DKK3 as a potential therapeutic approach for prostate cancer and highlight complexities in Dkk-3 regulation of TGF-β signaling

La eficacia a largo plazo de un programa de intervención basada en la atención plena (MBI) para la tartamudez: un estudio de caso

A mindfulness-based intervention (MBI) to address emotional factors that may negatively impact the fluency of the subjects was implemented. A MBI consisting of 5 weekly 2-h sessions was designed and applied to two stuttering cases. The goals were to lower stuttering and to reduce anxiety through the acquisition of mindfulness skills. Participants were asked to video-record a speaking task and to fill out anxiety and mindfulness skills self-reports pre and post-intervention. Both participants diminished stuttering error rates from severe (participant 1) and moderate (participant 2) to mild. Participant 2, who showed high pre-treatment anxiety, showed a significant reduction. A year-follow-up revealed that therapeutic gains in anxiety, mindfulness skills, and stuttering were successfully maintained in both participants. This is the first study focused on emotional aspects of stuttering showing long-term improvements through a MBI. MBI programs could serve as a promising complement for stuttering treatments.Se utilizó una intervención basada en mindfulness (MBI) para abordar los factores emocionales que pueden afectar negativamente a la fluidez de los sujetos a través de un estudio de caso. Se diseñó un MBI que constaba de 5 sesiones semanales de 2 horas y se aplicó a dos casos de tartamudez. Los objetivos eran dismunuir la tartamudez y reducir los síntomas de ansiedad mediante la adquisición de habilidades de atención plena. Se pidió a los participantes que grabaran en video una tarea oral y que cumplimentaran los autoinformes de ansiedad y de habilidades de atención plena antes y después de la intervención. En ambos participantes disminuyó el índice de errores de tartamudeo de grave (participante 1) y moderado (participante 2) a leve. El participante 2, que mostró una elevada ansiedad previa al tratamiento, mostró una reducción significativa. Al año de seguimiento los beneficios terapéuticos en ansiedad, habilidades de atención plena y tartamudeo se mantuvieron con éxito en ambos participantes. Se trata del primer estudio centrado en los aspectos emocionales de la tartamudez que muestra mejoras a largo plazo a través de un MBI. Por lo tanto, los programas MBI podrían servir como complemento prometedor a los tratamientos de tartamudezJennifer E. Moreno-Jiménez was granted a predoctoral fellowship, FPI-UAM 2017 of the Universidad Autónoma de Madri

Psychometric properties of the Spanish validation of the children's empathic attitudes questionnaire (CEAQ)

The prevalence of antisocial behavior in school settings is still discouraging. Students that often engage in aggressive acts may lack in the ability to appreciate the emotional consequences of their behaviors and share others’ emotions. The Children’s Empathic Attitudes Questionnaire (CEAQ) is one of the questionnaires used to assess empathy in children and early adolescents. This study is aimed to validate the Spanish version of CEAQ. The sample comprised 297 children (50% males), aged from 7 to 12 years (M = 9.53, SD = 1.2), from Madrid. Confirmatory factor analysis indicated an excellent fit for a unidimensional model, χ2(89) = 110.702, p = .059; CFI = .972; RMSEA = .029, 95% CI [.000, .045]. Multigroup invariance analysis showed no significant gender-related differences in all levels. Results also referred an acceptable reliability (ω = .824, r = .610). These results provide psychometric support for the use of the Spanish version of CEAQ as a valid and reliable instrument to assess empathy in children and youth population, especially for school-based intervention

Autofluorescence of stingray skeletal cartilage: hyperspectral imaging as a tool for histological characterization

Tessellated cartilage is a distinctive composite tissue forming the bulk of the skeleton of cartilaginous fishes (e.g. sharks and rays), built from unmineralized cartilage covered at the surface by a thin layer of mineralized tiles called tesserae. The finescale structure and composition of elasmobranch tessellated cartilage has largely been investigated with electron microscopy, micro-computed tomography and histology, but many aspects of tissue structure and composition remain uncharacterized. In our study, we demonstrate that the tessellated cartilage of a stingray exhibits a strong and diverse autofluorescence, a native property of the tissue which can be harnessed as an effective label-free imaging technique. The autofluorescence signal was excited using a broad range of wavelengths in confocal and light sheet microscopy, comparing several sample preparations (fresh; demineralized and paraffin-embedded; non-demineralized and plastic-embedded) and imaging the tissue at different scales. Autofluorescence varied with sample preparation with the signal in both plastic- and paraffin-embedded samples strong enough to allow visualization of finescale (≥ 1 μm) cellular and matrix structures, such as cell nuclei and current and former mineralization fronts, identifiable by globular mineralized tissue. A defined pericellular matrix (PCM) surrounding chondrocytes was also discernible, described here for the first time in elasmobranchs. The presence of a PCM suggests similarities with mammalian cartilage regarding how chondrocytes interact with their environment, the PCM in mammals acting as a transducer for biomechanical and biochemical signals. A posterior analysis of hyperspectral images by an MCR-ALS unmixing algorithm allowed identification of several distinct fluorescence signatures associated to specific regions in the tissue. Some fluorescence signatures identified could be correlated with collagen type II, the most abundant structural molecule of cartilage. Other fluorescence signatures, however, remained unidentified, spotlighting tissue regions that deserve deeper characterization and suggesting the presence of molecules still unidentified in elasmobranch skeletal cartilage. Our results show that autofluorescence can be a powerful exploratory imaging tool for characterizing less-studied skeletal tissues, such as tessellated cartilage. The images obtained are largely comparable with more commonly used techniques, but without the need for complicated sample preparations or external staining reagents standard in histology and electron microscopy (TEM, SEM)

Crew Scheduling for Netherlands Railways: "destination: customer"

: In this paper we describe the use of a set covering model with additional constraints for scheduling train drivers and conductors for the Dutch railway operator NS Reizigers. The schedules were generated according to new rules originating from the project "Destination: Customer" ("Bestemming: Klant" in Dutch). This project is carried out by NS Reizigers in order to increase the quality and the punctuality of its train services. With respect to the scheduling of drivers and conductors, this project involves the generation of efficient and acceptable duties with a high robustness against the transfer of delays of trains. A key issue for the acceptability of the duties is the included amount of variation per duty. The applied set covering model is solved by dynamic column generation techniques, Lagrangean relaxation and powerful heuristics. The model and the solution techniques are part of the TURNI system, which is currently used by NS Reizigers for carrying out several analyses concerning the required capacities of the depots. The latter are strongly influenced by the new rules

Timescales of Quartz Crystallization and the Longevity of the Bishop Giant Magma Body

Supereruptions violently transfer huge amounts (100 s–1000 s km3) of magma to the surface in a matter of days and testify to the existence of giant pools of magma at depth. The longevity of these giant magma bodies is of significant scientific and societal interest. Radiometric data on whole rocks, glasses, feldspar and zircon crystals have been used to suggest that the Bishop Tuff giant magma body, which erupted ∼760,000 years ago and created the Long Valley caldera (California), was long-lived (>100,000 years) and evolved rather slowly. In this work, we present four lines of evidence to constrain the timescales of crystallization of the Bishop magma body: (1) quartz residence times based on diffusional relaxation of Ti profiles, (2) quartz residence times based on the kinetics of faceting of melt inclusions, (3) quartz and feldspar crystallization times derived using quartz+feldspar crystal size distributions, and (4) timescales of cooling and crystallization based on thermodynamic and heat flow modeling. All of our estimates suggest quartz crystallization on timescales of <10,000 years, more typically within 500–3,000 years before eruption. We conclude that large-volume, crystal-poor magma bodies are ephemeral features that, once established, evolve on millennial timescales. We also suggest that zircon crystals, rather than recording the timescales of crystallization of a large pool of crystal-poor magma, record the extended periods of time necessary for maturation of the crust and establishment of these giant magma bodies

Modelling the tumour microenvironment in long-term microencapsulated 3D co-cultures recapitulates phenotypic features of disease progression

3D cell tumour models are generated mainly in non-scalable culture systems, using bioactive scaffolds. Many of these models fail to reflect the complex tumour microenvironment and do not allow long-term monitoring of tumour progression. To overcome these limitations, we have combined alginate microencapsulation with agitation-based culture systems, to recapitulate and monitor key aspects of the tumour microenvironment and disease progression. Aggregates of MCF-7 breast cancer cells were microencapsulated in alginate, either alone or in combination with human fibroblasts, then cultured for 15 days. In co-cultures, the fibroblasts arranged themselves around the tumour aggregates creating distinct epithelial and stromal compartments. The presence of fibroblasts resulted in secretion of pro-inflammatory cytokines and deposition of collagen in the stromal compartment. Tumour cells established cell–cell contacts and polarised around small lumina in the interior of the aggregates. Over the culture period, there was a reduction in oestrogen receptor and membranous E-cadherin alongside loss of cell polarity, increased collective cell migration and enhanced angiogenic potential in co-cultures. These phenotypic alterations, typical of advanced stages of cancer, were not observed in the mono-cultures of MCF-7 cells. The proposed model system constitutes a new tool to study tumour-stroma crosstalk, disease progression and drug resistance mechanisms

Імуногістохімічне виявлення судинного епітеліального ростового фактору в корі великих півкуль головного мозку при порушеннях кровообігу

Порушення кровопостачання мозку – одне з актуальних питань сучасної медицини, що обумовлено, як тяжкістю наслідків кожного конкретного випадку хвороби, так і рівнем показників захворюваності, що сягають пандемії, а смертність від цієї патології становить понад 20% і займає друге місце після серцево-судинних захворювань. Сьогодні зміни при ішемії мозку розглядаються як складний багатовекторний процес зі специфічною кінетикою на перебіг якого можна впливати, а не як одноманітну подію, як вважалось ще 20 років тому