Spatial perception in stereoscopic augmented reality based on multifocus sensing.

In many areas ranging from medical imaging to visual entertainment, 3D information acquisition and display is a key task. In this regard, in multifocus computational imaging, stacks of images of a certain 3D scene are acquired under different focus configurations and are later combined by means of post-capture algorithms based on image formation model in order to synthesize images with novel viewpoints of the scene. Stereoscopic augmented reality devices, through which is possible to simultaneously visualize the three dimensional real world along with overlaid digital stereoscopic image pair, could benefit from the binocular content allowed by multifocus computational imaging. Spatial perception of the displayed stereo pairs can be controlled by synthesizing the desired point of view of each image of the stereo-pair along with their parallax setting. The proposed method has the potential to alleviate the accommodation-convergence conflict and make augmented reality stereoscopic devices less vulnerable to visual fatigue.

Real-time phase retrieval in division of aperture microscopy with the transport of intensity equation.

The transport of intensity equation (TIE) allows to recover the phase of a microscopy sample from differently focused intensity measures along the axial direction of its optical field. In the present work, we propose a cost-effective technique for snapshot phase retrieval with TIE. The optics of a commercially available camera is replaced with a doublet system consisting of a microscope objective and a lenslet array with an extra lens mask attached to it. The system allows to obtain, in real-time and with no mechanical shift of either the sample or the sensor, the in-focus as well as a defocused image of the sample. From these two sub-aperture images, the intensity derivative term in TIE can then be approximated after image rectification. Phase is then retrieved for static as well as dynamic samples over the common view area. Validation experiments are presented.

Generalized Hough transform for 3D object recognition and visualization in integral imaging.

Object recognition is an automated image processing application of great interest in areas ranging from defect inspection to robot vision. In this regard, the generalized Hough transform is a well-established technique for the recognition of geometrical features even when they are partially occluded or corrupted by noise. To extend the original algorithm-aimed at detecting 2D geometrical features out of single images-we propose the robust integral generalized Hough transform, which corresponds to transformation under the generalized Hough transform of an elemental image array obtained from a 3D scene under integral imaging capture. The proposed algorithm constitutes a robust approach to pattern recognition in 3D scenes that takes into account information obtained not only from the individual processing of each image of the array but also from the spatial restrictions arising from perspective shifts between images. The problem of global detection of a 3D object of given size, position, and orientation is then exchanged under the robust integral generalized Hough transform for a more easily solved maximum detection in an accumulation (Hough) space dual to the elemental image array of the scene. Detected objects can then be visualized following refocusing schemes of integral imaging. Validation experiments for the detection and visualization of partially occluded 3D objects are presented. To the best of our knowledge, this is the first implementation of the generalized Hough transform for 3D object detection in integral imaging.

In vitro longitudinal lumbar spinal cord preparations to study sensory and recurrent motor microcircuits of juvenile mice.

In vitro spinal cord preparations have been extensively used to study microcircuits involved in the control of movement. By allowing precise control of experimental conditions coupled with state-of-the-art genetics, imaging, and electrophysiological techniques, isolated spinal cords from mice have been an essential tool in detailing the identity, connectivity, and function of spinal networks. The majority of the research has arisen from in vitro spinal cords of neonatal mice, which are still undergoing important postnatal maturation. Studies from adults have been attempted in transverse slices, however, these have been quite challenging due to the poor motoneuron accessibility and viability, as well as the extensive damage to the motoneuron dendritic trees. In this work, we describe two types of coronal spinal cord preparations with either the ventral or the dorsal horn ablated, obtained from mice of different postnatal ages, spanning from preweaned to 1 mo old. These semi-intact preparations allow recordings of sensory-afferent and motor-efferent responses from lumbar motoneurons using whole cell patch-clamp electrophysiology. We provide details of the slicing procedure and discuss the feasibility of whole cell recordings. The in vitro dorsal and ventral horn-ablated spinal cord preparations described here are a useful tool to study spinal motor circuits in young mice that have reached the adult stages of locomotor development.NEW & NOTEWORTHY In the past 20 years, most of the research into the mammalian spinal circuitry has been limited to in vitro preparations from embryonic and neonatal mice. We describe two in vitro longitudinal lumbar spinal cord preparations from juvenile mice that allow the study of motoneuron properties and respective afferent or efferent spinal circuits through whole cell patch clamp. These preparations will be useful to those interested in the study of microcircuits at mature stages of motor development.

Cdk5-Dependent Phosphorylation of CaV3.2 T-Type Channels: Possible Role in Nerve Ligation-Induced Neuropathic Allodynia and the Compound Action Potential in Primary Afferent C Fibers.

Voltage-gated T-type Ca2+ (CaV3) channels regulate diverse physiological events, including neuronal excitability, and have been linked to several pathological conditions such as absence epilepsy, cardiovascular diseases, and neuropathic pain. It is also acknowledged that calcium/calmodulin-dependent protein kinase II and protein kinases A and C regulate the activity of T-type channels. Interestingly, peripheral nerve injury induces tactile allodynia and upregulates CaV3.2 channels and cyclin-dependent kinase 5 (Cdk5) in dorsal root ganglia (DRG) and spinal dorsal horn. Here, we report that recombinant CaV3.2 channels expressed in HEK293 cells are regulatory targets of Cdk5. Site-directed mutagenesis showed that the relevant sites for this regulation are residues S561 and S1987. We also found that Cdk5 may regulate CaV3.2 channel functional expression in rats with mechanical allodynia induced by spinal nerve ligation (SNL). Consequently, the Cdk5 inhibitor olomoucine affected the compound action potential recorded in the spinal nerves, as well as the paw withdrawal threshold. Likewise, Cdk5 expression was upregulated after SNL in the DRG. These findings unveil a novel mechanism for how phosphorylation may regulate CaV3.2 channels and suggest that increased channel activity by Cdk5-mediated phosphorylation after SNL contributes nerve injury-induced tactile allodynia.SIGNIFICANCE STATEMENT Neuropathic pain is a current public health challenge. It can develop as a result of injury or nerve illness. It is acknowledged that the expression of various ion channels can be altered in neuropathic pain, including T-type Ca2+ channels that are expressed in sensory neurons, where they play a role in the regulation of cellular excitability. The present work shows that the exacerbated expression of Cdk5 in a preclinical model of neuropathic pain increases the functional expression of CaV3.2 channels. This finding is relevant for the understanding of the molecular pathophysiology of the disease. Additionally, this work may have a substantial translational impact, since it describes a novel molecular pathway that could represent an interesting therapeutic alternative for neuropathic pain.

Using a variant of coverslip hypoxia to visualize tumor cell alterations at increasing distances from an oxygen source.

Early stages in tumor development involve growth in confined spaces, where oxygen diffusion is limited and metabolic waste products accumulate. This hostile microenvironment imposes strong selective pressures on tumor cells, leading eventually to the survival and expansion of aggressive subclones that condition further tumor evolution. To model features of this microenvironment in vitro, a diffusional barrier can be introduced in the form of a coverslip placed on top of cells, a method termed coverslip hypoxia. Using a variant of this method, with larger volume between coverslip and cells and with oxygen diffusion occurring only through a small hole in the center of the coverslip, we have visualized alterations in LNCaP tumor cells as a function of their distance to the oxygen source at the center. We observed remarkable morphological changes in LNCaP cells as the distance from the center increases, with cells becoming highly spread, displaying dynamic membrane protrusions and occasionally adopting a migratory phenotype. Concomitantly, cells farther from the center displayed marked increases in the hypoxia marker hypoxyprobe, whereas extracellular pH decreased in the same direction. Cells with altered morphology displayed prominent increases in fibrillar actin, as well as swollen mitochondria with distorted cristae and accumulation of neutral lipid-containing intracellular vesicles. These results show that an in vitro microenvironment that models diffusional barriers encountered by tumors in situ can have profound effects on tumor cells. The coverslip hypoxia variant we describe can be used to characterize in vitro the response of tumor cells to environmental conditions that play crucial roles in early tumor development.

Fully invariant generalized Hough transform by out-of-focus multiview sensing with pupil array.

Recognition of geometrical shapes in real-time and fully invariant (i.e., invariant under changes in position, scale, and orientation) is a demanding task in automated image analysis. In particular, the generalized Hough transform (GHT) is a well-known algorithm for the recognition of complex patterns out of edge binary images even with disconnected boundaries or corrupted by noise. In this work we present a space multiplexed optical implementation of the GHT which, by exploiting the redundancy derived from multiview sensing of a two-dimensional image and its out- of-focus capture with an adequate pupil array, allows us to obtain in a single shot the GHT of this image invariant to target shift, scale, and orientation. Experimental validation of the working principle is presented, along with an assessment of the robustness of the system against noise in the input.

Specialized mechanoreceptor systems in rodent glabrous skin.

KEY POINTS: An ex vivo preparation was developed to record from single sensory fibres innervating the glabrous skin of the mouse forepaw. The density of mechanoreceptor innervation of the forepaw glabrous skin was found to be three times higher than that of hindpaw glabrous skin. Rapidly adapting mechanoreceptors that innervate Meissner's corpuscles were severalfold more responsive to slowly moving stimuli in the forepaw compared to those innervating hindpaw skin. We found a distinct group of small hairs in the centre of the mouse hindpaw glabrous skin that were exclusively innervated by directionally sensitive D-hair receptors. The directional sensitivity, but not the end-organ anatomy, were the opposite to D-hair receptors in the hairy skin. Glabrous skin hairs in the hindpaw are not ubiquitous in rodents, but occur in African and North American species that diverged more than 65 million years ago. ABSTRACT: Rodents use their forepaws to actively interact with their tactile environment. Studies on the physiology and anatomy of glabrous skin that makes up the majority of the forepaw are almost non-existent in the mouse. Here we developed a preparation to record from single sensory fibres of the forepaw and compared anatomical and physiological receptor properties to those of the hindpaw glabrous and hairy skin. We found that the mouse forepaw skin is equipped with a very high density of mechanoreceptors; >3 times more than hindpaw glabrous skin. In addition, rapidly adapting mechanoreceptors that innervate Meissner's corpuscles of the forepaw were severalfold more sensitive to slowly moving mechanical stimuli compared to their counterparts in the hindpaw glabrous skin. All other mechanoreceptor types as well as myelinated nociceptors had physiological properties that were invariant regardless of which skin area they occupied. We discovered a novel D-hair receptor innervating a small group of hairs in the middle of the hindpaw glabrous skin in mice. These glabrous skin D-hair receptors were direction sensitive albeit with an orientation sensitivity opposite to that described for hairy skin D-hair receptors. Glabrous skin hairs do not occur in all rodents, but are present in North American and African rodent species that diverged more than 65 million years ago. The function of these specialized hairs is unknown, but they are nevertheless evolutionarily very ancient. Our study reveals novel physiological specializations of mechanoreceptors in the glabrous skin that likely evolved to facilitate tactile exploration.

Fourier domain post-acquisition aperture reshaping from a multi-focus stack.

Computational optical imaging methods allow the extension of the functionality of traditional cameras. The shape of the aperture in an optical system determines the shape in which the out-of-focus points are blurred in a captured image. In this work we present a method in the Fourier domain that allows us, from an acquired multi-focus image stack, to synthesize images of a three-dimensional scene as if they had been acquired with apertures with arbitrary shapes. Partially extended depth-of-field as well as all-in-focus image reconstruction can be obtained as particular cases.

Reconstruction of perspective shifts and refocusing of a three-dimensional scene from a multi-focus image stack.

The convergence of optical imaging acquisition and image processing algorithms is a fast-evolving interdisciplinary research field focused on the reconstruction of images with novel features of interest. We propose a method for post-capture perspective shift reconstruction (in the x, y, and z directions) of a three-dimensional scene as well as refocusing with apertures of arbitrary shapes and sizes from an optimal multi-focus image stack. The approach is based on the reorganization of the acquired visual information considering a depth-variant point-spread function, which allows it to be applied to strongly defocused multi-focus image stacks. Our method is performed without estimating the depth map or segmenting the in-focus regions. A conventional camera combined with an electrically tunable lens is used for image acquisition and does not require scale transformation or registration between the acquired images. Experimental results for both real and synthetic data images are provided and compared to state-of-the-art schemes.

Image segmentation by nonlinear filtering of optical Hough transform.

The identification and extraction (i.e., segmentation) of geometrical features is crucial in many tasks requiring image analysis. We present a method for the optical segmentation of features of interest from an edge enhanced image. The proposed method is based on the nonlinear filtering (implemented by the use of a spatial light modulator) of the generalized optical Hough transform and is capable of discriminating features by shape and by size. The robustness of the method against noise in the input, low contrast, or overlapping of geometrical features is assessed, and experimental validation of the working principle is presented.

All-in-focus image reconstruction under severe defocus.

Limited depth-of-focus is a problem in many fields of optics, e.g., microscopy and macro-photography. We propose a new physically based method with a space variant point spread function (PSF) to accomplish all-in-focus reconstruction (image fusion) from a multi-focus image sequence in order to extend the depth-of-field. The proposed method works well under strong defocus conditions for color image stacks of arbitrary length. Experimental results are provided to demonstrate that our method outperforms state-of-the-art image fusion algorithms for strong defocus on both synthetic as well as real data images.

Optical implementation of the generalized Hough transform with totally incoherent light.

The generalized Hough transform is a well-established technique for detecting complex shapes in images containing noisy or missing data. We present an efficient optical implementation of this transform using an electrical lens with variable focal length and a rotating pupil mask matching the pattern to be found. The proposed setup works under fully (i.e., both spatially and temporally) incoherent illumination and can handle orientation changes or scale variations in the pattern. Validation experiments showing its real-time application are presented.

Real-time pattern recognition using an optical generalized Hough transform.

We present some pattern recognition applications of a generalized optical Hough transform and the temporal multiplexing strategies for dynamic scale and orientation-variant detection. Unlike computer-based implementations of the Hough transform, in principle its optical implementation does not impose restrictions on the execution time or on the resolution of the images or frame rate of the videos to be processed, which is potentially useful for real-time applications. Validation experiments are presented.

Differential effects of coverslip-induced hypoxia and cobalt chloride mimetic hypoxia on cellular stress, metabolism, and nuclear structure.

Hypoxia has profound effects on cell physiology, both in normal or pathological settings like cancer. In this study, we asked whether a variant of coverslip-induced hypoxia that recapitulates the conditions found in the tumor microenvironment would elicit similar cellular responses compared to the well established model of cobalt chloride-induced hypoxia. Comparable levels of nuclear HIF-1α were observed after 24 h of coverslip-induced hypoxia or cobalt chloride treatment in CAL-27 oral squamous carcinoma cells. However, cellular stress levels assessed by reactive oxygen species production and lipid droplet accumulation were markedly increased in coverslip-induced hypoxia compared to cobalt chloride treatment. Conversely, mitochondrial ATP production sharply decreased after coverslip-induced hypoxia but was preserved in the presence of cobalt chloride. Coverslip-induced hypoxia also had profound effects in nuclear organization, assessed by changes in nuclear dry mass distribution, whereas these effects were much less marked after cobalt chloride treatment. Taken together, our results show that coverslip-induced hypoxia effects on cell physiology and structure are more pronounced than mimetic hypoxia induced by cobalt chloride treatment. Considering also the simplicity of coverslip-induced hypoxia, our results therefore underscore the usefulness of this method to recapitulate in vitro the effects of hypoxic microenvironments encountered by cells in vivo.

Sensory Schwann cells set perceptual thresholds for touch and selectively regulate mechanical nociception.

Previous work identified nociceptive Schwann cells that can initiate pain. Consistent with the existence of inherently mechanosensitive sensory Schwann cells, we found that in mice, the mechanosensory function of almost all nociceptors, including those signaling fast pain, were dependent on sensory Schwann cells. In polymodal nociceptors, sensory Schwann cells signal mechanical, but not cold or heat pain. Terminal Schwann cells also surround mechanoreceptor nerve-endings within the Meissner's corpuscle and in hair follicle lanceolate endings that both signal vibrotactile touch. Within Meissner´s corpuscles, two molecularly and functionally distinct sensory Schwann cells positive for Sox10 and Sox2 differentially modulate rapidly adapting mechanoreceptor function. Using optogenetics we show that Meissner's corpuscle Schwann cells are necessary for the perception of low threshold vibrotactile stimuli. These results show that sensory Schwann cells within diverse glio-neural mechanosensory end-organs are sensors for mechanical pain as well as necessary for touch perception.

[Factors related to sleep quality in Emergency Medical Technicians and their sleep habits during COVID-19 pandemic.] / Factores relacionados con la calidad de sueño de los técnicos en emergencias sanitarias y sus hábitos de sueño durante la pandemia de COVID-19.

OBJECTIVE: Emergency Medical Technicians (EMTs) show a high prevalence of sleep problems. Adding to these problems, another factor appeared two years ago: the COVID-19 pandemic. The objectives of this study were to describe the sleep quality and habits in a sample of EMTs in Spain during COVID-19 pandemic considering the factors related to them. METHODS: A national cross-sectional study was carried out in Spain between October 2020 and February 2021. EMTs who worked in basic and advanced life support ambulances were invited to participate in an online survey. Several sociodemographic variables were analyzed, as well as sleep quality, stress symptoms, pain, food consumption and physical activity. Logistic regression models were used to examine the associations between these variables and sleep quality. RESULTS: 340 EMTs participated in the study. Of them, 59.4% had poor sleep quality and the habits of 32.2% got worse during the pandemic. The EMTs with a poor sleep quality presented in higher proportion stress symptoms (OR: 4.19; 95% CI: 2.16-8.11; p<0.001), pain (OR: 3.19; 95% CI: 1.7-6.01; p<0.001) and a weekly consumption of sugar-sweetened beverages greater than two glasses (OR: 3.6; 95% CI: 1.86-6.98; p<0.001). CONCLUSIONS: EMTs show a high prevalence of poor sleep quality, which got worse during the pandemic. The factors related to them are stress, pain and consumption of sugar-sweetened beverages. This study provides important information so that the emergency services can develop health promotion programs focused on these professionals.

OBJETIVO: Los Técnicos en Emergencias Sanitarias (TES) presentan una alta prevalencia de problemas de sueño. A estos problemas se sumó otro factor hace dos años: la pandemia de la COVID-19. Los objetivos de este estudio fueron describir la calidad y hábitos de sueño en una muestra de TES en España durante la pandemia, teniendo en cuenta sus factores relacionados. METODOS: Se realizó un estudio transversal de carácter nacional, que fue desarrollado en España entre octubre de 2020 y febrero de 2021. Los TES que trabajaban en ambulancias de Soporte Vital Básico o Avanzado fueron invitados a participar en un cuestionario online. Se analizaron varias variables sociodemográficas, la calidad del sueño, los síntomas de estrés, el dolor, el consumo alimentario y la actividad física. Se utilizaron modelos de regresión logística para examinar las asociaciones entre estas variables y la calidad de sueño. RESULTADOS: Participaron 340 TES en el estudio. La calidad de sueño del 59,4% era mala y los hábitos de sueño del 32,2% empeoraron durante la pandemia. Aquellos con un sueño de mala calidad presentaban en mayor proporción síntomas de estrés (OR: 4,19; IC 95%: 2,16-8,11; p<0,001), dolor (OR: 3,19; IC 95%: 1,7-6,01; p<0,001) y un consumo semanal de bebidas azucaradas superior a dos vasos semanales (OR: 3,6; IC 95%: 1,86-6,98; p<0,001). CONCLUSIONES: Los TES presentan una alta prevalencia de mala calidad de sueño, habiendo empeorado además durante la pandemia. Entre sus factores relacionados se encuentran el estrés, el dolor y el consumo de bebidas azucaradas. Este estudio aporta información importante para que los servicios de emergencias puedan desarrollar programas de promoción de la salud centrados en estos profesionales.

Contributions of viral oncogenes of HPV-18 and hypoxia to oxidative stress and genetic damage in human keratinocytes.

Infection with high-risk human papillomaviruses like HPV-16 and HPV-18 is highly associated with the development of cervical and other cancers. Malignant transformation requires viral oncoproteins E5, E6 and E7, which promote cell proliferation and increase DNA damage. Oxidative stress and hypoxia are also key factors in cervical malignant transformation. Increased levels of reactive species of oxygen (ROS) and nitrogen (RNS) are found in the hypoxic tumor microenvironment, promoting genetic instability and invasiveness. In this work, we studied the combined effect of E5, E6 and E7 and hypoxia in increasing oxidative stress and promoting DNA damage and nuclear architecture alterations. HaCaT cells containing HPV-18 viral oncogenes (HaCaT E5/E6/E7-18) showed higher ROS levels in normoxia and higher levels of RNS in hypoxia compared to HaCaT parental cells, as well as higher genetic damage in hypoxia as measured by γH2AX and comet assays. In hypoxia, HaCaT E5/E6/E7-18 increased its nuclear dry mass and both cell types displayed marked heterogeneity in nuclear dry mass distribution and increased nuclear foci. Our results show contributions of both viral oncogenes and hypoxia to oxidative stress, DNA damage and altered nuclear architecture, exemplifying how an altered microenvironment combines with oncogenic transformation to promote tumor progression.

Color encoding of binary fringes for gamma correction in 3-D profiling.

Three-dimensional profiling by sinusoidal fringe projection using PSI-algorithms are distorted by the nonlinear response of digital cameras and commercial video projectors. To solve the problem, we present a fringe generation technique that consists of projecting and acquiring a temporal sequence of strictly binary color patterns, whose (adequately weighted) average leads to sinusoidal fringe patterns with the required number of bits, which allows for a reliable three-dimensional profile using a PSI-algorithm. Validation experiments are presented.

Edge enhancement of color images using a digital micromirror device.

A method for orientation-selective enhancement of edges in color images is proposed. The method utilizes the capacity of digital micromirror devices to generate a positive and a negative color replica of the image used as input. When both images are slightly displaced and imagined together, one obtains an image with enhanced edges. The proposed technique does not require a coherent light source or precise alignment. The proposed method could be potentially useful for processing large image sequences in real time. Validation experiments are presented.