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- Normative mice retinal thickness: 16-month longitudinal characterization of wild-type mice and changes in a model of Alzheimer's diseasePublication . Batista, Ana; Guimarães, Pedro; Martins, João; Moreira, Paula I.; Ambrósio, António F.; Castelo-Branco, Miguel; Serranho, Pedro; Bernardes, RuiAnimal models of disease are paramount to understand retinal development, the pathophysiology of eye diseases, and to study neurodegeneration using optical coherence tomography (OCT) data. In this study, we present a comprehensive normative database of retinal thickness in C57BL6/129S mice using spectral-domain OCT data. The database covers a longitudinal period of 16 months, from 1 to 16 months of age, and provides valuable insights into retinal development and changes over time. Our findings reveal that total retinal thickness decreases with age, while the thickness of individual retinal layers and layer aggregates changes in different ways. For example, the outer plexiform layer (OPL), photoreceptor inner segments (ILS), and retinal pigment epithelium (RPE) thickened over time, whereas other retinal layers and layer aggregates became thinner. Additionally, we compare the retinal thickness of wild-type (WT) mice with an animal model of Alzheimer's disease (3×Tg-AD) and show that the transgenic mice exhibit a decrease in total retinal thickness compared to age-matched WT mice, with statistically significant differences observed at all evaluated ages. This normative database of retinal thickness in mice will serve as a reference for future studies on retinal changes in neurodegenerative and eye diseases and will further our understanding of the pathophysiology of these conditions.
- Characterization of the retinal changes of the 3×Tg-AD mouse model of Alzheimer’s diseasePublication . Ferreira, Hugo; Martins, João; Nunes, Ana; Moreira, Paula I.; Castelo-Branco, Miguel; Ambrósio, António F.; Serranho, Pedro; Bernardes, RuiAlzheimer’s disease (AD) is a progressive neurodegenerative disorder whose diagnosis remains a notable challenge. The literature suggests that cerebral changes precede AD symptoms by over two decades, implying a significantly advanced stage of AD by the time it is usually diagnosed. In the study herein, texture analysis was applied to computed optical coherence tomography ocular fundus images to identify differences between a group of the transgenic mouse model of the Alzheimer’s disease (3×Tg-AD) and a group of wild-type mice, at the ages of one and two-months-old. A substantial difference between groups was found at both time-points across all neuroretina’s layers. Here, the inner nuclear layer stands out both in the level of statistically significant differences and on the extension of these differences which span through the imaged area. Also, the progression of AD is suggested to be spotted by texture analysis as demonstrated by the significant difference found in the inner plexiform and the outer nuclear layers from the age of one to the age of two-months-old. These findings demonstrate the potential of the use of the retina and texture analysis to the diagnosis of AD and monitor AD progression. Besides, the differences between groups found in this study suggest that the 3×Tg-AD model may be inappropriate to study early changes associated with the AD and other animal models should be tested following the same path and rationale. Moreover, these results also suggest that the human genes present in these transgenic mice may have an impact on the neurodevelopment of offspring which would justify the significant changes found at the age of one-month-old.
- A hybrid method for sound-hard obstacle reconstructionPublication . Kress, Rainer; Serranho, PedroWe are interested in solving the inverse problem of acoustic wave scattering to reconstruct the position and the shape of sound-hard obstacles from a given incident field and the corresponding far field pattern of the scattered field. The method we suggest is an extension of the hybrid method for the reconstruction of sound-soft cracks as presented in [R. Kress, P. Serranho, A hybrid method for two-dimensional crack reconstruction, Inverse Problems 21 (2005) 773–784] to the case of sound-hard obstacles. The designation of the method is justified by the fact that it can be interpreted as a hybrid between a regularized Newton method applied to a nonlinear operator equation with the operator that maps the unknown boundary onto the solution of the direct scattering problem and a decomposition method in the spirit of the potential method as described in [A. Kirsch, R. Kress, On an integral equation of the first kind in inverse acoustic scattering, in: Cannon, Hornung (Eds.), Inverse Problems, ISNM, vol. 77, 1986, pp. 93–102. Since the method does not require a forward solver for each Newton step its computational costs are reduced. By some numerical examples we illustrate the feasibility of the method.
- Synthetic volume from real optical coherence tomography dataPublication . Serranho, Pedro; Bernardes, Rui; Maduro, Cristina; Santos, Torcato; Cunha-Vaz, JoséPurpose: To build a mathematical model to mimic a real OCT b-scan/volume without noise, in order to establish a ground truth for image processing performance metrics. Methods: Current image processing techniques (eg. despeckling filtering methods) with application to optical coherence tomography (OCT) rely on the respective qualitative evaluation of its results. Quantitative approaches are reduced to using synthetic images which consists of an homogeneous background and a set of abstract objects, eg. cubes and spheres. In this work, we suggest a mathematical model to address this issue by creating a synthetic b-scan/volume based on any real OCT data scan, that can be used as ground truth for processing methods testing. Eye scans of healthy volunteers and eyes of patients with age-related macular degeneration and diabetic retinopathy were used following Cirrus OCT (Carl Zeiss Meditec, Dublin, CA, USA) scans using both the 200x200x1024 and the 512x128x1024 Macular Cube Protocols. Each of these eye scans was processed in order to extract required parameters. For healthy subjects, only the segmentation of the inner limiting membrane (ILM) and the retinal pigment epithelium (RPE) are needed, though for pathologic eyes, the segmentation of other structures to be preserved might also be needed. In each segmented region, OCT data is surface fitted using appropriate basis functions. Results: A total of 45 scans were processed resulting in the synthetic data representing the major characteristics of the respective real OCT scans. These results have been used for testing the performance of an improved complex diffusion despeckling method proposed by some of the authors. Conclusions: This process allows to automatically compute a synthetic OCT scan mimicking a real one. In this way, this process makes it possible to quantify the result of any processing (eg. filtering) by providing adequate synthetic data as ground truth.
- Bioestatística com SPSS: notas de apoioPublication . Serranho, Pedro; Ramos, Maria do RosárioNotas de apoio ao curso de "Biostestatística com SPSS" leccionado pelos Prof. Doutor Francisco Caramelo (Universidade de Coimbra), Prof. Doutora Maria Rosário Ramos e Prof. Doutor Pedro Serranho.
- A hybrid method for two-dimensional crack reconstructionPublication . Kress, Rainer; Serranho, PedroWe present a new method for solving the time-harmonic inverse scattering problem for sound-soft or perfectly conducting cracks in two dimensions. Our approach extends a method that was recently suggested by one of us for inverse obstacle scattering. It can be viewed as a hybrid between a regularized Newton iterationmethod applied to a nonlinear operator equation involving the operator that, for a fixed incident wave, maps the crack onto the far-field pattern of the scattered wave and a decomposition method due to Kirsch and Kress. As an important feature, in contrast to the traditional Newton iterations for solving inverse scattering problems, our method does not require a forward solver for each iteration step. The theoretical background of the method is based on the minimization of a cost function containing an additional penalty term to deal with reconstructing the full crack. Numerical examples illustrate the feasibility of the method and its stability with respect to noisy data. We expect that the method can also be extended to sound-hard cracks.
- Two-dimensional segmentation of the retinal vascular network from optical coherence tomographyPublication . Rodrigues, Pedro; Guimarães, Pedro; Santos, Torcato; Simão, Sílvia; Miranda, Telmo; Serranho, Pedro; Bernardes, RuiThe automatic segmentation of the retinal vascular network from ocular fundus images has been performed by several research groups. Although different approaches have been proposed for traditional imaging modalities, only a few have addressed this problem for optical coherence tomography (OCT). Furthermore, these approaches were focused on the optic nerve head region. Compared to color fundus photography and fluorescein angiography, two-dimensional ocular fundus reference images computed from three-dimensional OCT data present additional problems related to system lateral resolution, image contrast, and noise. Specifically, the combination of system lateral resolution and vessel diameter in the macular region renders the process particularly complex, which might partly explain the focus on the optic disc region. In this report, we describe a set of features computed from standard OCT data of the human macula that are used by a supervised-learning process (support vector machines) to automatically segment the vascular network. For a set of macular OCT scans of healthy subjects and diabetic patients, the proposed method achieves 98% accuracy, 99% specificity, and 83% sensitivity. This method was also tested on OCT data of the optic nerve head region achieving similar results.
- Progressive changes in pneumococcal carriage in children attending daycare in Portugal after 6 years of gradual conjugate vaccine introduction show falls in most residual vaccine serotypes but no net replacement or trends in diversityPublication . Rodrigues, Fernanda; Foster, Dona; Caramelo, Francisco; Serranho, Pedro; Gonçalves, Guilherme; Januário, Luís; Finn, AdamObjectives: To track ongoing trends in pneumococcal (Sp) serotype carriage under the selection pressure of moderate pneumococcal conjugate vaccine (PCV) use, children in a community in Portugal were studied in the same months in 3 consecutive years. Methods: Nasopharyngeal specimens were collected (children aged 3 months to <7 years) in 8 urban daycare centers in February 2008 (n = 561) and 2009 (n = 585). Sp isolates were serotyped. Results: While demographics were similar in 2008–2009 and a previously reported sample in 2007, PCV coverage (at least one dose) in the children studied rose from 76.5% to 84% although national coverage was lower than this. Sp carriage fell from 61% to 51% with a concomitant fall in PCV7 serotype carriage from 12.1% to 4.3%. Remaining PCV7 serotypes declined to near (23F) or totally (6B, 14) undetectable levels except 19F which persisted unchanged in around 4% of children. Although carriage of 3 and 6C rose, there was no net increase in non-PCV7 serotypes and no progressive trend in serotype diversity. Conclusions: Ecological changes induced by PCVs where uptake is moderate appear to be different from high usage settings. We report falling Sp carriage due to PCV7 serotype disappearance with persistence of 19F and no ongoing net replacement after several years of PCV7 use and slowly rising uptake.
- Retinal aging in 3× Tg-AD mice model of Alzheimer's diseasePublication . Guimarães, Pedro; Serranho, Pedro; Martins, João; Moreira, Paula I.; Ambrósio, António Francisco; Castelo-Branco, Miguel; Bernardes, RuiThe retina, as part of the central nervous system (CNS), can be the perfect target for in vivo, in situ, and noninvasive neuropathology diagnosis and assessment of therapeutic efficacy. It has long been established that several age-related brain changes are more pronounced in Alzheimer’s disease (AD). Nevertheless, in the retina such link is still under-explored. This study investigates the differences in the aging of the CNS through the retina of 3×Tg-AD and wild-type mice. A dedicated optical coherence tomograph imaged mice’s retinas for 16 months. Two neural networks were developed to model independently each group’s ages and were then applied to an independent set containing images fromboth groups. Our analysis shows amean absolute error of 0.875±1.1×10−2 and 1.112 ± 1.4 × 10−2 months, depending on training group. Our deep learning approach appears to be a reliable retinal OCT aging marker. We show that retina aging is distinct in the two classes: the presence of the three mutated human genes in the mouse genome has an impact on the aging of the retina. For mice over 4 months-old, transgenic mice consistently present a negative retina age-gap when compared to wildtype mice, regardless of training set. This appears to contradict AD observations in the brain. However, the ‘black-box” nature of deep-learning implies that one cannot infer reasoning. We can only speculate that some healthy age-dependent neural adaptations may be altered in transgenic animals.
- Noninvasive evaluation of retinal leakage using optical coherence tomographyPublication . Bernardes, Rui; Santos, Torcato; Serranho, Pedro; Lobo, Conceição; Cunha-Vaz, JoséPurpose: To demonstrate the association between changes in the blood-retinal barrier (BRB) identified by fluorescein leakage and those in the optical properties of the human retina determined by optical coherence tomography (OCT) and show how these changes can be quantified and their location identified within the retina. Methods: Two imaging techniques were applied: the retinal leakage analyzer, to map BRB function into intact or disrupted regions, and OCT, to measure refractive index changes along the light path within the human ocular fundus. Results: A total of 140 comparisons were made, 77 between areas of regions receiving the same classification (intact or disrupted BRB) and 63 between areas of regions receiving distinct classifications, from 4 pathological cases: 2 eyes with nonproliferative diabetic retinopathy and 2 eyes with wet age-related macular degeneration. In all cases, the distribution of OCT data between regions of intact and regions of disrupted BRB, identified by the retinal leakage analyzer, was quantified and was statistically significantly different (p < 0.001). In addition, it was found that the differences could be localized in the retina to specific structural sequences. Conclusions: Using a novel method to analyze OCT data, we showed that it may be possible to quantify differences in the extracellular compartment in eyes with retinal disease and alterations of the BRB. Based on quantitative techniques, our findings demonstrate the presence of indirect information on the BRB status within noninvasive OCT data.