Multispectral fundus analysis

Purpose - To generate a reflectance model of the fundus that allows an accurate non-invasive quantification of blood and pigments. Methods - A Monte Carlo simulation was used to produce a mathematical model of light interaction with the fundus at different wavelengths. The model predictions were compared with fundus images from normal volunteers in several spectral bands (peaks at 507, 525, 552, 585, 596 and 611nm). Th e model was then used to calculate the concentration and distribution of the known absorbing components of the fundus. Results - The shape of the statistical distribution of the image data generally corresponded to that of the model data; the model however appears to overestimate the reflectance of the fundus in the longer wavelength region.As the absorption by xanthophyll has no significant eff ect on light transport above 534nm, its distribution in the fundus was quantified: the wavelengths where both shape and distribution of image and model data matched (<553nm) were used to train a neural network which was then applied to every point in the image data. The xanthophyll distribution thus found was in agreement with published literature data in normal subjects. Conclusion - We have developed a method for optimising multi-spectral imaging of the fundus and a computer image analysis capable of estimating information about the structure and properties of the fundus. Th e technique successfully calculates the distribution of xanthophyll in the fundus of healthy volunteers. Further improvement of the model is required to allow the deduction of other parameters from images; investigations in known pathology models are also necessary to establish if this method is of clinical use in detecting early chroido-retinopathies, hence providing a useful screening and diagnostic tool.

Multi-frequency segmental bio-impedance device:design, development and applications

Bio-impedance analysis (BIA) provides a rapid, non-invasive technique for body composition estimation. BIA offers a convenient alternative to standard techniques such as MRI, CT scan or DEXA scan for selected types of body composition analysis. The accuracy of BIA is limited because it is an indirect method of composition analysis. It relies on linear relationships between measured impedance and morphological parameters such as height and weight to derive estimates. To overcome these underlying limitations of BIA, a multi-frequency segmental bio-impedance device was constructed through a series of iterative enhancements and improvements of existing BIA instrumentation. Key features of the design included an easy to construct current-source and compact PCB design. The final device was trialled with 22 human volunteers and measured impedance was compared against body composition estimates obtained by DEXA scan. This enabled the development of newer techniques to make BIA predictions. To add a ‘visual aspect’ to BIA, volunteers were scanned in 3D using an inexpensive scattered light gadget (Xbox Kinect controller) and 3D volumes of their limbs were compared with BIA measurements to further improve BIA predictions. A three-stage digital filtering scheme was also implemented to enable extraction of heart-rate data from recorded bio-electrical signals. Additionally modifications have been introduced to measure change in bio-impedance with motion, this could be adapted to further improve accuracy and veracity for limb composition analysis. The findings in this thesis aim to give new direction to the prediction of body composition using BIA. The design development and refinement applied to BIA in this research programme suggest new opportunities to enhance the accuracy and clinical utility of BIA for the prediction of body composition analysis. In particular, the use of bio-impedance to predict limb volumes which would provide an additional metric for body composition measurement and help distinguish between fat and muscle content.

The value of magnetoencephalography to guide electrode implantation in epilepsy

Purpose: To investigate if magnetoencephalography (MEG) can identify implantation sites for intracranial recordings (IR). Method: Two groups of 12 patients assessed for surgery with IR with and without MEG were compared (MEG and control groups). In the control group, non-invasive presurgical assessment without MEG suggested clear hypotheses for implantation. In the MEG group, non-invasive assessment was inconclusive, and MEG was used to identify implantation sites. Both groups were matched for implantation type. The success of implantation was defined by findings in IR: a) Focal seizure onset; b)Unilateral focal abnormal responses to single pulse electrical stimulation(SPES); and c) Concordance between a) and b). Results: In all MEG patients, at least one virtual MEG electrode generated suitable hypotheses for the location of implantations. The proportion of patients showing focal seizure onset restricted to one hemisphere was similar in control and MEG groups (6/12 vs. 11/12, Fisher’s exact test,p = 0.0686). The proportion of patients showing unilateral responses to SPES was lower in the control than in the MEG group (7/12 vs. 12/12,p = 0.0373). Conclusion: The MEG group showed similar or higher incidence of successful implantations than controls.

Modelling the growth of young individuals of Pinus nigra on thin carbonate soils under climate change in Hungary

Climate change highly impacts on tree growth and also threatens the forest of the karstic terrains. From the 1980s the frequency of decay events of the Pinus nigra Arnold forests showed a marked increase in Hungary. To understanding the vulnerability of Pinus nigra forests to climate change on shallow karstic soils in continental-sub Mediterranean climatic conditions we developed the study of three sampled population in the typical karstic landscape of Veszprém in North Transdanubia. We built our model on non-invasive approach using the annual growth of the individuals. MPI Echam5 climate model and as aridity index the Thornthwaite Agrometeorological Index were used. Our results indicate that soil thickness up to 11 cm has a major influence on the main growth intensity, however, aridity determines the annual growth rate. Our model results showed that the increasing decay frequency in the last decades was a parallel change to the decreasing growth rate of pines. The climate model predicts the similar, increased decay frequency to the presents. Our results can be valid for a wider areas of the periphery of Mediterranean climate zone while the annual-growth based model is a cost-effective and simple method to study the vitality of pine trees in a given area.

Micropartículas de quitosana estruturadas com aerosil®: estabilidade, adsorção, encapsulação e liberação de substâncias ativas

Biodegradable microspheres used as controlled release systems are important in pharmaceutics. Chitosan biopolymer represents an attractive biomaterial alternative because of its physicochemical and biological characteristics. Chitosan microspheres are expected to become promising carrier systems for drug and vaccine delivery, especially for non-invasive ways oral, mucosal and transdermal routes. Controlling the swelling rate and swelling capacity of the hydrogel and improving the fragile nature of microspheres under acidic conditions are the key challenges that need to be overcomed in order to enable the exploration of the full pharmaceutical potential use of these microparticles. Many studies have focused on the modification of chitosan microsphere structures with cross-linkers, various polymers blends and new organic-inorganic hybrid systems in order to obtain improved properties. In this work, microspheres made of chitosan and nanosized hydrophobic silica (Aerosil R972) were produced by a method consisting of two steps. First, a preparation of a macroscopically homogeneous chitosan-hydrophobic silica dispersion was prepared followed by spray drying. FTIR spectroscopy, X-ray powder diffraction, differential scanning calorimetry, thermal gravimetric analysis, scanning electron microscopy (SEM) and high-resolution transmission electron microscopy (TEM) were used to characterize the microspheres. Also, the were conducted acid stability, moisture sorption capacity, release properties and biological assays. The chitosan-hydrophobic silica composite microspheres showed improved thermal degradation, lower water affinity, better acid stability and ability to retard rifampicin and propranolol hydrochloride (drug models) release under simulated physiological conditions. In vitro biocompatibility studies indicated low cytotoxicity and low capacity to activate cell production of the pro-inflammatory mediator nitric oxide. The results show here encourage further studies on the use of the new chitosan-hydrophobic silica composite microspheres as drug carrier systems via oral or nasal routes.

Prospective Estimation of Radiation Dose and Image Quality for Optimized CT Performance

X-ray computed tomography (CT) is a non-invasive medical imaging technique that generates cross-sectional images by acquiring attenuation-based projection measurements at multiple angles. Since its first introduction in the 1970s, substantial technical improvements have led to the expanding use of CT in clinical examinations. CT has become an indispensable imaging modality for the diagnosis of a wide array of diseases in both pediatric and adult populations [1, 2]. Currently, approximately 272 million CT examinations are performed annually worldwide, with nearly 85 million of these in the United States alone [3]. Although this trend has decelerated in recent years, CT usage is still expected to increase mainly due to advanced technologies such as multi-energy [4], photon counting [5], and cone-beam CT [6].

Despite the significant clinical benefits, concerns have been raised regarding the population-based radiation dose associated with CT examinations [7]. From 1980 to 2006, the effective dose from medical diagnostic procedures rose six-fold, with CT contributing to almost half of the total dose from medical exposure [8]. For each patient, the risk associated with a single CT examination is likely to be minimal. However, the relatively large population-based radiation level has led to enormous efforts among the community to manage and optimize the CT dose.

As promoted by the international campaigns Image Gently and Image Wisely, exposure to CT radiation should be appropriate and safe [9, 10]. It is thus a responsibility to optimize the amount of radiation dose for CT examinations. The key for dose optimization is to determine the minimum amount of radiation dose that achieves the targeted image quality [11]. Based on such principle, dose optimization would significantly benefit from effective metrics to characterize radiation dose and image quality for a CT exam. Moreover, if accurate predictions of the radiation dose and image quality were possible before the initiation of the exam, it would be feasible to personalize it by adjusting the scanning parameters to achieve a desired level of image quality. The purpose of this thesis is to design and validate models to quantify patient-specific radiation dose prospectively and task-based image quality. The dual aim of the study is to implement the theoretical models into clinical practice by developing an organ-based dose monitoring system and an image-based noise addition software for protocol optimization.

More specifically, Chapter 3 aims to develop an organ dose-prediction method for CT examinations of the body under constant tube current condition. The study effectively modeled the anatomical diversity and complexity using a large number of patient models with representative age, size, and gender distribution. The dependence of organ dose coefficients on patient size and scanner models was further evaluated. Distinct from prior work, these studies use the largest number of patient models to date with representative age, weight percentile, and body mass index (BMI) range.

With effective quantification of organ dose under constant tube current condition, Chapter 4 aims to extend the organ dose prediction system to tube current modulated (TCM) CT examinations. The prediction, applied to chest and abdominopelvic exams, was achieved by combining a convolution-based estimation technique that quantifies the radiation field, a TCM scheme that emulates modulation profiles from major CT vendors, and a library of computational phantoms with representative sizes, ages, and genders. The prospective quantification model is validated by comparing the predicted organ dose with the dose estimated based on Monte Carlo simulations with TCM function explicitly modeled.

Chapter 5 aims to implement the organ dose-estimation framework in clinical practice to develop an organ dose-monitoring program based on a commercial software (Dose Watch, GE Healthcare, Waukesha, WI). In the first phase of the study we focused on body CT examinations, and so the patient’s major body landmark information was extracted from the patient scout image in order to match clinical patients against a computational phantom in the library. The organ dose coefficients were estimated based on CT protocol and patient size as reported in Chapter 3. The exam CTDIvol, DLP, and TCM profiles were extracted and used to quantify the radiation field using the convolution technique proposed in Chapter 4.

With effective methods to predict and monitor organ dose, Chapters 6 aims to develop and validate improved measurement techniques for image quality assessment. Chapter 6 outlines the method that was developed to assess and predict quantum noise in clinical body CT images. Compared with previous phantom-based studies, this study accurately assessed the quantum noise in clinical images and further validated the correspondence between phantom-based measurements and the expected clinical image quality as a function of patient size and scanner attributes.

Chapter 7 aims to develop a practical strategy to generate hybrid CT images and assess the impact of dose reduction on diagnostic confidence for the diagnosis of acute pancreatitis. The general strategy is (1) to simulate synthetic CT images at multiple reduced-dose levels from clinical datasets using an image-based noise addition technique; (2) to develop quantitative and observer-based methods to validate the realism of simulated low-dose images; (3) to perform multi-reader observer studies on the low-dose image series to assess the impact of dose reduction on the diagnostic confidence for multiple diagnostic tasks; and (4) to determine the dose operating point for clinical CT examinations based on the minimum diagnostic performance to achieve protocol optimization.

Chapter 8 concludes the thesis with a summary of accomplished work and a discussion about future research.

Imputing accelerometer non-wear time when assessing children's moderate-to-vigorous physical activity

BACKGROUND: Moderate-to-vigorous physical activity (MVPA) is an important determinant of children’s physical health, and is commonly measured using accelerometers. A major limitation of accelerometers is non-wear time, which is the time the participant did not wear their device. Given that non-wear time is traditionally discarded from the dataset prior to estimating MVPA, final estimates of MVPA may be biased. Therefore, alternate approaches should be explored. OBJECTIVES: The objectives of this thesis were to 1) develop and describe an imputation approach that uses the socio-demographic, time, health, and behavioural data from participants to replace non-wear time accelerometer data, 2) determine the extent to which imputation of non-wear time data influences estimates of MVPA, and 3) determine if imputation of non-wear time data influences the associations between MVPA, body mass index (BMI), and systolic blood pressure (SBP). METHODS: Seven days of accelerometer data were collected using Actical accelerometers from 332 children aged 10-13. Three methods for handling missing accelerometer data were compared: 1) the “non-imputed” method wherein non-wear time was deleted from the dataset, 2) imputation dataset I, wherein the imputation of MVPA during non-wear time was based upon socio-demographic factors of the participant (e.g., age), health information (e.g., BMI), and time characteristics of the non-wear period (e.g., season), and 3) imputation dataset II wherein the imputation of MVPA was based upon the same variables as imputation dataset I, plus organized sport information. Associations between MVPA and health outcomes in each method were assessed using linear regression. RESULTS: Non-wear time accounted for 7.5% of epochs during waking hours. The average minutes/day of MVPA was 56.8 (95% CI: 54.2, 59.5) in the non-imputed dataset, 58.4 (95% CI: 55.8, 61.0) in imputed dataset I, and 59.0 (95% CI: 56.3, 61.5) in imputed dataset II. Estimates between datasets were not significantly different. The strength of the relationship between MVPA with BMI and SBP were comparable between all three datasets. CONCLUSION: These findings suggest that studies that achieve high accelerometer compliance with unsystematic patterns of missing data can use the traditional approach of deleting non-wear time from the dataset to obtain MVPA measures without substantial bias.

Implementação de classificador de tarefas mentais baseado em EEG

BARBOSA, André F. ; SOUZA, Bryan C. ; PEREIRA JUNIOR, Antônio ; MEDEIROS, Adelardo A. D.de, . Implementação de Classificador de Tarefas Mentais Baseado em EEG. In: CONGRESSO BRASILEIRO DE REDES NEURAIS, 9., 2009, Ouro Preto, MG. Anais... Ouro Preto, MG, 2009

Implementação de classificador de tarefas mentais baseado em EEG

BARBOSA, André F. ; SOUZA, Bryan C. ; PEREIRA JUNIOR, Antônio ; MEDEIROS, Adelardo A. D.de, . Implementação de Classificador de Tarefas Mentais Baseado em EEG. In: CONGRESSO BRASILEIRO DE REDES NEURAIS, 9., 2009, Ouro Preto, MG. Anais... Ouro Preto, MG, 2009

Revisiting molecular diagnostics of Streptococcus pneumoniae

Streptococcus pneumoniae is a human pathobiont that colonizes the nasopharynx. S. pneumoniae is responsible for causing non-invasive and invasive disease such as otitis, pneumonia, meningitis, and sepsis, being a leading cause of infectious diseases worldwide. Due to similarities with closely related species sharing the same niche, it may be a challenge to correctly distinguish S. pneumoniae from its relatives when using only non-culture based methods such as real time PCR (qPCR). In 2007, a molecular method targeting the major autolysin (lytA) of S. pneumoniae by a qPCR assay was proposed by Carvalho and collaborators to identify pneumococcus. Since then, this method has been widely used worldwide. In 2013, the gene encoding for the ABC iron transporter lipoprotein PiaA, was proposed by Trzcinzki and collaborators to be used in parallel with the lytA qPCR assay. However, the presence of lytA gene homologues has been described in closely related species such as S. pseudopneumoniae and S. mitis and the presence of piaA gene is not ubiquitous between S. pneumoniae. The hyaluronate lyase gene (hylA) has been described to be ubiquitous in S. pneumoniae. This gene has not been used so far as a target for the identification of S. pneumoniae. The aims of our study were to evaluate the specificity, sensitivity, positive predicted value (PPV) and negative predicted value (NPV) of the lytA and piaA qPCR methods; design and implement a new assay targeting the hylA gene and evaluate the same parameters above described; analyze the assays independently and the possible combinations to access what is the best approach using qPCR to identify S. pneumoniae. A total of 278 previously characterized strains were tested: 61 S. pseudopneumoniae, 37 Viridans group strains, 30 type strains from other streptococcal species and 150 S. pneumoniae strains. The collection included both carriage and disease isolates. By Mulilocus Sequence Analysis (MLSA) we confirmed that strains of S. pseudopneumoniae could be misidentified as S. pneumoniae when lytA qPCR assay is used. The results showed that as a single target, lytA had the best combination of specificity, sensitivity, PPV and NPV being, 98.5%, 100.0%, 98.7% and 100.0% respectively. The combination of targets with the best values of specificity, sensibility, PPV and NPV were lytA and piaA, with 100.0%, 93.3%, 97.9% and 92.6%, respectively. Nonetheless by MLSA we confirmed that strains of S. pseudopneumoniae could be misidentified as S. pneumoniae and some capsulated (23F, 6B and 11A) and non-capsulated S. pneumoniae were not Identified using this assay. The hylA gene as a single target had the lowest PPV. Nonetheless it was capable to correctly identify all S. pneumoniae.

Lógica borrosa para la estimación de estados críticos de una pila de combustible PEM

La determinación en tiempo real de los estados críticos de operación de la pila de combustible de membrana intercambio protónico (siglas en ingles, PEM) es uno de los principales retos para los sistemas de control de pilas de combustible PEM. En este trabajo, se presenta el desarrollo e implementación de un método no invasivo de bajo coste basado en técnicas de decisión borrosa que permite estimar los estados críticos de operación de la pila de combustible PEM. La estimación se realiza mediante perturbaciones al estado de operación de la pila y el análisis posterior de la evolución temporal del voltaje generado por la pila. La implementación de esta técnica de estimulación-percepción de estado de la pila de combustible para la detección de estados críticos constituye una novedad y un paso hacia el control autónomo en óptimas condiciones de la operación de las pilas de combustible PEM.

Detección temprana del cáncer de mama mediante la termografía en Ecuador

En los últimos reportes de Quito-Ecuador ocupa el primer lugar en diagnósticos de cáncer en las mujeres. Las campañas de concienciación y las investigaciones han permitido mejorar el diagnóstico y su tratamiento, pero en países de recursos bajos y medios aun es un problema sin resolver, debido a varios factores: diagnóstico en fases avanzadas y recursos insuficientes que ayuden en su pronta detección. Se conoce que en Ecuador por ejemplo, cerca de cuatro de cada diez mujeres diagnosticadas de cáncer de mama lo son en estadios avanzados (III y IV) cuyo tratamiento es costoso y complejo. Actualmente, según la OMS el reto para todos los países es detectar tempranamente la enfermedad. Por tanto, se debe abordar nuevos retos, tales como la búsqueda de nuevas herramientas y tecnologías costo efectivas que ofrezcan ventajas al actual método, que está basado en mamografías y auto examen. El uso de nuevas tecnologías permitirá identificar la enfermedad en estadios tempranos, lo que dará lugar a que la sobrevida libre de enfermedad aumente, los costos de la atención disminuyan y el pronóstico vital sea mayor. Este trabajo propone incorporar el uso de la termografía como complementaria al diagnóstico precoz de bajo coste y no invasiva.

In vivo microvascular oximetry using multispectral imaging

This thesis describes the application of multispectral imaging to several novel oximetry applications. Chapter 1 motivates optical microvascular oximetry, outlines oxygen transport in the body, describes the theory of oximetry, and describes the challenges associated with in vivo oximetry, in particular imaging through tissue. Chapter 2 reviews various imaging techniques for quantitative in vivo oximetry of the microvasculature, including multispectral and hyperspectral imaging, photoacoustic imaging, optical coherence tomography, and laser speckle techniques. Chapter 3 describes a two-wavelength oximetry study of two microvascular beds in the anterior segment of the eye: the bulbar conjunctival and episcleral microvasculature. This study reveals previously unseen oxygen diffusion from ambient air into the bulbar conjunctival microvasculature, altering the oxygen saturation of the bulbar conjunctiva. The response of the bulbar conjunctival and episcleral microvascular beds to acute mild hypoxia is quantified and the rate at which oxygen diffuses into bulbar conjunctival vessels is measured. Chapter 4 describes the development and application of a highly novel non-invasive retinal angiography technique: Oximetric Ratio Contrast Angiography (ORCA). ORCA requires only multispectral imaging and a small perturbation of blood oxygen saturation to produce angiographic sequences. A pilot study of ORCA in human subjects was conducted. This study demonstrates that ORCA can produce angiographic sequences with features such as sequential vessel filling and laminar flow. The application and challenges of ORCA are discussed, with emphasis on comparison with other angiography techniques, such as fluorescein angiography. Chapter 5 describes the development of a multispectral microscope for oximetry in the spinal cord dorsal vein of rats. Measurements of blood oxygen saturation are made in the dorsal vein of both healthy rats, and in rats with the Experimental autoimmune encephalomyelitis (EAE) disease model of multiple sclerosis. The venous blood oxygen saturation of EAE disease model rats was found to be significantly lower than that of healthy controls, indicating increased oxygen uptake from blood in the EAE disease model of multiple sclerosis. Chapter 6 describes the development of video-rate red eye oximetry; a technique which could enable stand-off oximetry of the blood-supply of the eye with high temporal resolution. The various challenges associated with video-rate red eye oximetry are investigated and their influence quantified. The eventual aim of this research is to track circulating deoxygenation perturbations as they arrive in both eyes, which could provide a screening method for carotid artery stenosis, which is major risk-factor for stroke. However, due to time constraints, it was not possible to thoroughly investigate if video-rate red eye can detect such perturbations. Directions and recommendations for future research are outlined.

Shaping surface acoustic waves for cardiac tissue engineering

The heart is a non-regenerating organ that gradually suffers a loss of cardiac cells and functionality. Given the scarcity of organ donors and complications in existing medical implantation solutions, it is desired to engineer a three-dimensional architecture to successfully control the cardiac cells in vitro and yield true myocardial structures similar to native heart. This thesis investigates the synthesis of a biocompatible gelatin methacrylate hydrogel to promote growth of cardiac cells using biotechnology methodology: surface acoustic waves, to create cell sheets. Firstly, the synthesis of a photo-crosslinkable gelatin methacrylate (GelMA) hydrogel was investigated with different degree of methacrylation concentration. The porous matrix of the hydrogel should be biocompatible, allow cell-cell interaction and promote cell adhesion for growth through the porous network of matrix. The rheological properties, such as polymer concentration, ultraviolet exposure time, viscosity, elasticity and swelling characteristics of the hydrogel were investigated. In tissue engineering hydrogels have been used for embedding cells to mimic native microenvironments while controlling the mechanical properties. Gelatin methacrylate hydrogels have the advantage of allowing such control of mechanical properties in addition to easy compatibility with Lab-on-a-chip methodologies. Secondly in this thesis, standing surface acoustic waves were used to control the degree of movement of cells in the hydrogel and produce three-dimensional engineered scaffolds to investigate in-vitro studies of cardiac muscle electrophysiology and cardiac tissue engineering therapies for myocardial infarction. The acoustic waves were characterized on a piezoelectric substrate, lithium niobate that was micro-fabricated with slanted-finger interdigitated transducers for to generate waves at multiple wavelengths. This characterization successfully created three-dimensional micro-patterning of cells in the constructs through means of one- and two-dimensional non-invasive forces. The micro-patterning was controlled by tuning different input frequencies that allowed manipulation of the cells spatially without any pre- treatment of cells, hydrogel or substrate. This resulted in a synchronous heartbeat being produced in the hydrogel construct. To complement these mechanical forces, work in dielectrophoresis was conducted centred on a method to pattern micro-particles. Although manipulation of particles were shown, difficulties were encountered concerning the close proximity of particles and hydrogel to the microfabricated electrode arrays, dependence on conductivity of hydrogel and difficult manoeuvrability of scaffold from the surface of electrodes precluded measurements on cardiac cells. In addition, COMSOL Multiphysics software was used to investigate the mechanical and electrical forces theoretically acting on the cells. Thirdly, in this thesis the cardiac electrophysiology was investigated using immunostaining techniques to visualize the growth of sarcomeres and gap junctions that promote cell-cell interaction and excitation-contraction of heart muscles. The physiological response of beating of co-cultured cardiomyocytes and cardiac fibroblasts was observed in a synchronous and simultaneous manner closely mimicking the native cardiac impulses. Further investigations were carried out by mechanically stimulating the cells in the three-dimensional hydrogel using standing surface acoustic waves and comparing with traditional two-dimensional flat surface coated with fibronectin. The electrophysiological responses of the cells under the effect of the mechanical stimulations yielded a higher magnitude of contractility, action potential and calcium transient.

Comparação entre técnicas de aferição da temperatura corpórea em micos-estrela cativos (Callithrix penicillata) e sua relação com parâmetros hematológicos, peso corporal e umidade e temperatura do ambiente

Dissertação (mestrado)—Universidade de Brasília, Faculdade de Ciências da Saúde, Programa de Pós-Graduação em Ciências da Saúde, 2015.