Detecció, caracterització i estudi de les sibilàncies en senyals de so respiratori de pacients amb malalties pulmonars

L'aportació més important d'aquesta tesi doctoral és la proposta de noves tècniques de processament del senyal per a l'anàlisi objectiu d'uns tipus de sons respiratoris, anomenats sibilàncies, com a eina complementària pel diagnòstic mèdic i pel seguiment clínic de malalties pulmonars.Per a detectar les sibilàncies, inicialment s'han proposat i validat dos algorismes basats en l'anàlisi de l'espectrograma del senyal: el Shabtai-Musih de Mitjana Local (SMML) i l'Algorisme de Detecció de Sibilàncies Adaptatiu Local (LAWDA). La validació, realitzada a partir de 60 senyals de so amb l'ajut d'un metge especialista, ha mostrat una sensibilitat en la detecció de sibilàncies del 90.3% pel SMML i del 93.9% pel LAWDA en el tram de flux de l'espiració forçada comprès entre 1.2 i 0.2 l/s. Pel tram de flux de 0.2 a 0 l/s els valors han estat del 64.5% i del 71.0% respectivament.Per a l'estudi s'han fet servir senyals de so adquirits durant la maniobra espiromètrica que s'utilitza en la pràctica clínica habitual per a l'avaluació de la funció pulmonar de pacients. La base de dades conté 37 pacients, 15 de sans que formen el grup de control, 16 amb asma bronquial no fumadors i 6 que pateixen malaltia pulmonar obstructiva crònica (COPD). Per a cada pacient s'han estudiat un mínim de 6 maniobres: 3 en estat basal i 3 passats 20 minuts després de l'aplicació de 1 mg de Terbutalina (broncodilatador).S'ha realitzat un estudi estadístic de paràmetres característics del tram d'interès (1.2 a 0.2 l/s): el nombre de sibilàncies detectades, la freqüència mitjana de la sibilància de potència pic (FMSPPM) i mitjana (FMSPMM) màximes, el promig de les freqüències mitjanes de les sibilàncies detectades (PFM), i els percentatges de la maniobra ocupats per monofonia, polifonia o sense sibilàncies. S'han obtingut diferències significatives entre els grups per la mitjana i la desviació típica intrapacient d'alguns dels paràmetres estudiats tot i que els resultats no sempre coincidien per als dos algorismes. Cal destacar l'observació de diferències molt significatives entre els pacients de control i els asmàtics (pPer a estudiar la dinàmica de canvi de les vies aèries dels grups de pacients, també s'ha realitzat un estudi estadístic de les diferències entre les mitjanes i les desviacions típiques intrapacient dels paràmetres calculats abans i després d'aplicar un fàrmac broncodilatador. La majoria de paràmetres no han presentat diferències de comportament entre estats significatives en un mateix grup de pacients. També s'han definit els paràmetres de canvi en signe i valor absolut, que es defineixen com la resta entre els valors anteriors i els posteriors a l'aplicació del fàrmac. De l'estudi estadístic d'aquests paràmetres es conclou que hi ha diferències entre estats per un mateix pacient i que aquestes són de vegades significatives per diferenciar grups, sobretot quan s'estudien en valor absolut. Un cop validada la utilitat de la tècnica s'ha desenvolupat l'Algorisme de Detecció de Sibilàncies Genèric (GWDA), basat en la transformada de Malvar i el pseudoespectre. Aquest serveix per a detectar i estudiar les sibilàncies en altres maniobres i menors nivells de flux respiratori, i facilita la integració en dispositius de temps real. En la validació del nou algorisme s'ha obtingut una sensibilitat superior als altres tant pel tram comprès entre 1.2 i 0.2 l/s del flux (96.7 %) com pel tram de 0.2 a 0 l/s (100 %). S'ha realitzat l'estudi de l'espiració forçada utilitzant el GWDA i s'han estudiat estadísticament els diferents paràmetres calculats per les sibilàncies detectades pels pacients dels diferents grups. També s'han obtingut diferències molt significatives en estudiar el nombre de sibilàncies detectades en asmàtics i controls (pEls estudis estadístics s'han complementat en aquest cas amb un anàlisi discriminant per a avaluar la possibilitat de realitzar un sistema automàtic de classificació basat només en paràmetres provinents de les sibilàncies. L'anàlisi discriminant s'ha realitzat combinant fins a un màxim de 3 paràmetres que presentaven diferències significatives entre grups i ha mostrat taxes de classificació de fins al 91.9 % en la fase d'entrenament i del 77.8 % per la de test.Finalment, s'ha realitzat un estudi preliminar del nombre de sibilàncies detectades durant la maniobra de volum corrent per mitjà del GWDA.La aportación más importante de esta tesis doctoral es la propuesta de nuevas técnicas de procesado de señal para el análisis objetivo de un tipo de sonidos respiratorios, las sibilancias, como herramienta complementaria para el diagnostico médico y para seguimiento clínico de enfermedades pulmonares.Para detectar sibilancias, inicialmente se han propuesto y validado dos algoritmos basados en el análisis del espectrograma señal: el Shabtai-Musih de Media Local (SMML) y el Algoritmo de Detección de Sibilancias Adaptativo Local (LAWDA). La validación, realizada a partir de 60 señales de sonido con la ayuda de un médico especialista, ha mostrado una sensibilidad en la detección de sibilancias del 90.3% para el SMML y del 93.9% para el LAWDA en el tramo de flujo de la espiración forzada comprendido entre 1.2 i 0.2 l/s. Para el tramo de flujo de 0.2 a 0 l/s los valores han sido del 64.5% i del 71.0% respectivamente.Para el estudio se han utilizado señales de sonido adquiridos durante la maniobra espirométrica que se utiliza en la práctica clínica habitual para evaluar la función pulmonar de pacientes. La base de datos contiene 37 pacientes, 15 de sanos que forman el grupo de control, 16 con asma bronquial no fumadores y 6 que sufren enfermedad pulmonar obstructiva crónica (COPD). Para cada paciente se han estudiado un mínimo de 6 maniobras: 3 en estado basal y 3 pasados 20 minutos de la aplicación de 1 mg de Terbutalina (broncodilatador).Se ha realizado un estudio estadístico de parámetros característicos del tramo de interés (1.2 a 0.2 l/s): el número de sibilancias detectadas, la frecuencia media de la sibilancia de potencia pico (FMSPPM) y media (FMSPMM) máximas, el promedio de las frecuencias medias de las sibilancias detectadas (PFM), y los porcentajes de la maniobra ocupados por monofonía, polifonía o sin sibilancias. Se han obtenido diferencias significativas entre grupos para la media y la desviación típica intrapaciente de algunos de los parámetros estudiados aunque los resultados no siempre coincidían para los dos algoritmos. Cabe destacar la observación de diferencias muy significativas entre los pacientes de control y los asmáticos (pPara estudiar la dinámica de cambio de las vías aéreas de los grupos de pacientes, también se ha realizado un estudio estadístico de las diferencias entre las medias y las desviaciones típicas intrapaciente de los parámetros calculados antes y después de aplicar la Terbutalina. La mayoría de parámetros no han presentado diferencias de comportamiento entre estados significativos en un mismo grupo de pacientes. También se han definido los parámetros de cambio con signo y valor absoluto como la resta entre los valores anteriores y los posteriores a la aplicación del fármaco. Del estudio estadístico de estos parámetros se concluye que hay diferencias entre estados para un mismo paciente y que estas son a veces significativas para diferenciar grupos, sobretodo cuando se estudian en valor absoluto. Una vez validada la utilidad de la técnica se ha desarrollado el Algoritmo de Detección de Sibilancias Genérico (GWDA), basado en la transformada de Malvar y el pseudoespectro. Este sirve para detectar y estudiar las sibilancias en otras maniobras y menores niveles de flujo respiratorio, y facilita la integración en dispositivos de tiempo real. En la validación del nuevo algoritmo se ha obtenido una sensibilidad superior a los otros tanto para el tramo comprendido entre 1.2 i 0.2 l/s del flujo (96.7 %) como para el tramo de 0.2 a 0 l/s (100 %). Se ha realizado el estudio de la espiración forzada utilizando el GWDA y se han estudiado estadísticamente los parámetros calculados para las sibilancias detectadas en los pacientes de los diferentes grupos. También se han obtenido diferencias muy significativas al estudiar el número de sibilancias detectadas en asmáticos y controles (pLos estudios estadísticos se han completado en este caso con un análisis discriminante para evaluar la posibilidad de realizar un sistema automático de clasificación basado solamente en parámetros provenientes de las sibilancias. El análisis discriminante se ha realizado combinando hasta un máximo de 3 parámetros que presentaban diferencias significativas entre grupos y ha mostrado índices de clasificación de hasta el 91.9 % en la fase de entrenamiento y del 77.8 % para la de test.Finalmente, se ha realizado un estudio preliminar del número de sibilancias detectadas durante la maniobra de volumen corriente por medio del GWDA.The main contribution of this PhD Thesis is to propose new signal processing techniques to objectively analyze respiratory sounds, specifically wheezes, as a medical diagnostic complementary tool and to monitor the clinical evolution of pulmonary diseases.Initially, two algorithms based on the spectrogram analysis have been proposed and validated in order to detect wheezes: the Local Mean Shabtai-Musih (SMML) and the Local Adaptive Wheezes Detection Algorithm (LAWDA). Validation, done using 60 sound signals and with the assessment of an specialized medical doctor, shows a wheezes detection sensibility of 90.3% for the SMML and of 93.9% for the LAWDA when 1.2 to 0.2 l/s forced exhalation flow levels are evaluated. These values go down to 64.5% and 71.0% respectively when the 0.2 to 0 l/s segment is used instead. Sound signals used for this study were acquired from the patient during the spirometry maneuver that is used for the assessment of pulmonary function in regular clinical practice. Data base is composed by 37 individuals: 16 non-smoker bronchial asthma patients, 6 chronic obstructive pulmonary disease (COPD) affected individuals, and 15 control subjects. A minimum of 6 maneuvers have been studied for each patient: 3 in basal state and 3 after 20 minutes of applying a 1 mg dose of Terbutaline (bronchodilator). For the flow segment of interest (1.2 to 0.2 l/s), a statistical study of its characteristic parameters: number of detected wheezes, mean frequency of the wheeze with higher power peak (FMSPPM) or mean power (FMSPMM), the mean value of the detected wheezes mean frequencies (PFM), and the percentage of maneuver occupied by monophony, polyphony or without wheezes; has been performed. Significant differences between groups have been found for the intrapatient mean and standard deviation of some of the studied parameters but results were not always coincident for the two detection algorithms. A remarkable result has been encountered when studying the mean and standard deviation of the number of detected wheezes before and after applying a bronchodilator drug: very significant differences have been found (pIn order to study the patient groups airways dynamics changes, a statistical study of the differences between means and standard deviations of the parameters calculated before and after the application of the bronchodilator has been also performed. Most of the parameters did not show significant behavior differences between states of the same patients group. Also, the change parameters with sign and in absolute value have been defined. These are the results obtained by subtracting the values obtained before and after applying the drug. The statistical study of these parameters concludes that patients show differences between states and that these are sometimes significant in order to separate groups, especially when studied in absolute value. Once validated the utility of the technique the General Wheezes Detection Algorithm (GWDA) has been developed, based on the Malvar transform and the pseudospectrum. This algorithm is useful to detect and study wheezes on alternative maneuvers and lower respiratory flow levels, and facilitates the integration on real time devices. After validating it a higher sensitivity than the other algorithms has been found for the 1.2 to 0.2 l/s flow segment (96.7%) and also for the 0.2 to 0 l/s range (100%). The GWDA has been used to perform the forced exhalation study and the parameters obtained from the wheezes detected in each of the different groups patients have been studied statistically. Very significant differences have been also found when studying the number of detected wheezes in asthma and control groups (pFor the GWDA, the statistical studies have been completed with a discriminant analysis to evaluate the possibility to develop an automatic classification system only based on the parameters calculated from the detected wheezes. The discriminant analysis has been done combining up to a maximum of 3 parameters that showed significant differences between groups and classification rates of 91.9% for the training and of 77.8% for the testing patients sets have been accomplished.Finally, a preliminary study of the number of wheezes detected during the sporadic inspiration and exhalation maneuver has been done using the GWDA

Array of Microfluidic Beam Resonators for Density and Viscosity Analysis of Liquids

This paper reports on the design, fabrication, and evaluation of a mass density and viscosity sensor based on an array of polysilicon microbeam resonators integrated with 20 pL fluidic microchannels. When filled with water, resonators exhibit resonant frequencies close to 500 KHz and Q-factor values of 400 operating at atmospheric pressure and ambient temperature. Real-time measurements are highly reproducible and only require 250 μL of the sample fluid. The built-in interferometric readout enables automatic detection of the beams increasing the throughput analysis and reducing detection times. The frequency shift response shows a linear behavior in accordance with the density of evaluated solvents, organic solutions, and alcoholic drinks, reporting a mass responsivity of 7.4 Hz/pg. Also, the sensor is capable of measuring the viscosity of liquid phase samples with a resolution of 0.15 cP by tracking the Q-factor response of the sensor within a linear regime between 1 to 2.6 cP. This approach demonstrates the ability to identify in real-time changes of fluids in the liquid phase that could provide a valuable assessment for bioanalytical applications

Building of a flexible microfluidic plasmo-nanomechanical biosensor for live cell analysis

Biosensor devices can constitute an advanced tool for monitoring and study complex dynamic biological processes, as for example cellular adhesion. Cellular adhesion is a multipart process with crucial implications in physiology (i.e. immune response, tissue nature, architecture maintenance, or behaviour and expansion of tumor cells). This work focuses on offering a controlled methodology in order to fabricate a flexible plasmo-nanomechanical biosensor placed within a microfluidic channel as a new tool for future cell adhesion studies. We designed, fabricated, and optically and mechanically characterized this novel optical biosensor. As a proof-of-concept of its functionality, the biosensor was employed to observe fibroblasts adhesion in a cell culture. The device is configured by an hexagonal array of flexible rigid/soft polymeric nanopillars capped with plasmonic gold nanodisks integrated inside a microfluidic channel. The fabrication employs low-cost and large-scale replica molding techniques using two different polymers materials (EPOTECK OG142 and 310 M). By using those materials the spring constant of the polymer nanopillars (k) can be fabricated from 1.19E-02 [N/m] to 5.35E+00 [N/m] indicating different mechanical sensitivities to shear stress. Therefore, the biosensor has the feasibility to mimic soft and rigid tissues important for the description of cellular nanoscale behaviours. The biosensor exhibits a suitable bulk sensitivity of 164 nm or 206 nm/refractive index unit respectively, depending on the base material. The range of calculated forces goes from ≈1.98 nN to ≈.942 μN. This supports that the plasmo-nanomechanical biosensors could be employed as novel tool to study living cells behavior

Combined dielectrophoresis and impedance systems for bacteria analysis in microfluidic on-chip platforms

Bacteria concentration and detection is time-consuming in regular microbiology procedures aimed to facilitate the detection and analysis of these cells at very low concentrations. Traditional methods are effective but often require several days to complete. This scenario results in low bioanalytical and diagnostic methodologies with associated increased costs and complexity. In recent years, the exploitation of the intrinsic electrical properties of cells has emerged as an appealing alternative approach for concentrating and detecting bacteria. The combination of dielectrophoresis (DEP) and impedance analysis (IA) in microfluidic on-chip platforms could be key to develop rapid, accurate, portable, simple-to-use and cost-effective microfluidic devices with a promising impact in medicine, public health, agricultural, food control and environmental areas. The present document reviews recent DEP and IA combined approaches and the latest relevant improvements focusing on bacteria concentration and detection, including selectivity, sensitivity, detection time, and conductivity variation enhancements. Furthermore, this review analyses future trends and challenges which need to be addressed in order to successfully commercialize these platforms resulting in an adequate social return of public-funded investments

A label-free nanostructured plasmonic biosensor based on Blu-ray discs with integrated microfluidics for sensitive biodetection

Altres ajuts: CONACYT, Grant/Award Number: 225362Nanostructure-based plasmonic biosensors have quickly positioned themselves as interesting candidates for the design of portable optical biosensor platforms considering the potential benefits they can offer in integration, miniaturization, multiplexing, and real-time label-free detection. We have developed a simple integrated nanoplasmonic sensor taking advantage of the periodic nanostructured array of commercial Blu-ray discs. Sensors with two gold film thicknesses (50 and 100 nm) were fabricated and optically characterized by varying the oblique-angle of the incident light in optical reflectance measurements. Contrary to the use normal light incidence previously reported with other optical discs, we observed an enhancement in sensitivity and a narrowing of the resonant linewidths as the light incidence angle was increased, which could be related to the generation of Fano resonant modes. The new sensors achieve a figure of merit (FOM) up to 35 RIU and a competitive bulk limit of detection (LOD) of 6.3×10 RIU. These values significantly improve previously reported results obtained with normal light incidence reflectance measurements using similar structures. The sensor has been combined with versatile, simple, ease to-fabricate microfluidics. The integrated chip is only 1 cm (including a PDMS flow cell with a 50 µm height microfluidic channel fabricated with double-sided adhesive tape) and all the optical components are mounted on a 10 cm×10 cm portable prototype, illustrating its facile miniaturization, integration and potential portability. Finally, to assess the label-free biosensing capability of the new sensor, we have evaluated the presence of specific antibodies against the GTF2b protein, a tumor-associate antigen (TAA) related to colorectal cancer. We have achieved a LOD in the pM order and have assessed the feasibility of directly measuring biological samples such as human serum

Multiple biomarkers biosensor with just-in-time functionalization: Application to prostate cancer detection

We present a novel lab-on-a-chip (LOC) device for the simultaneous detection of multiple biomarkers using simple voltage measurements. The biosensor functionalization is performed in-situ, immediately before its use, facilitating reagents storage and massive devices fabrication. Sensitivity, limit of detection (LOD) and limit of quantification (LOQ) are tunable depending on the in-chip flown sample volumes. As a proof-of-concept, the system has been tested and adjusted to quantify two proteins found in blood that are susceptible to be used combined, as a screening tool, to diagnose prostate cancer (PCa): prostate-specific antigen (PSA) and spondin-2 (SPON2). This combination of biomarkers has been reported to be more specific for PCa diagnostics than the currently accepted but rather controversial PSA indicator. The range of detection for PSA and SPON2 could be adjusted to the clinically relevant range of 1 to 10. ng/ml. The system was tested for specificity to the evaluated biomarkers. This multiplex system can be modified and adapted to detect a larger quantity of biomarkers, or different ones, of relevance to other specific diseases.This research was partially supported by the Ministerio de Asuntos Exteriores y de Cooperación (MAEC) (MAEC-AECID, Convocatoria 2011-12) and by the Agencia Española de Cooperación Internacional (AECID) (586678 para el Programa II.E) with a scholarship. CIBER-BBN is an initiative funded by the VI National R&D&i Plan 2008–2011, Iniciativa Ingenio 2010, Consolider Program, CIBER Actions and financed by the Instituto de Salud Carlos III with assistance from the European Regional Development Fund. The Nanobioengineering group has support from the Commission for Universities and Research of the Department of Innovation, Universities, and Enterprise of the Generalitat de Catalunya (2009 SGR 505).Peer Reviewe

Array of microfluidic beam resonators for density and viscosity analysis of liquids

This paper reports on the design, fabrication, and evaluation of a mass density and viscosity sensor based on an array of polysilicon microbeam resonators integrated with 20 pL fluidic microchannels. When filled with water, resonators exhibit resonant frequencies close to 500 KHz and Q-factor values of 400 operating at atmospheric pressure and ambient temperature. Real-time measurements are highly reproducible and only require 250 μL of the sample fluid. The builtin interferometric readout enables automatic detection of the beams increasing the throughput analysis and reducing detection times. The frequency shift response shows a linear behavior in accordance with the density of evaluated solvents, organic solutions, and alcoholic drinks, reporting a mass responsivity of 7.4 Hz/pg. Also, the sensor is capable of measuring the viscosity of liquid phase samples with a resolution of 0.15 cP by tracking the Q-factor response of the sensor within a linear regime between 1 to 2.6 cP. This approach demonstrates the ability to identify in real-time changes of fluids in the liquid phase that could provide a valuable assessment for bioanalytical applications.This work was supported in part by the National Council for Science and Technology (CONACyT-Mexico) and also with the Integrated Clean Room for Micro and Nano Fabrication of the Centro Nacional de Microelectróica - Instituto de Microelectrónica de Barcelona (ICTS IMB-CNM); in part by the Departament d’Universitats, Recerca i Societat de la Informació de la Generalitat de Catalunya, under Grant 2014SGR624; and in part by the Severo Ochoa Centers of Excellence Program of Spanish MINECO under Grant SEV-2013-0295.Peer Reviewe

Array of Microfluidic Beam Resonators for Density and Viscosity Analysis of Liquids

This paper reports on the design, fabrication, and evaluation of a mass density and viscosity sensor based on an array of polysilicon microbeam resonators integrated with 20 pL fluidic microchannels. When filled with water, resonators exhibit resonant frequencies close to 500 KHz and Q-factor values of 400 operating at atmospheric pressure and ambient temperature. Real-time measurements are highly reproducible and only require 250 μL of the sample fluid. The built-in interferometric readout enables automatic detection of the beams increasing the throughput analysis and reducing detection times. The frequency shift response shows a linear behavior in accordance with the density of evaluated solvents, organic solutions, and alcoholic drinks, reporting a mass responsivity of 7.4 Hz/pg. Also, the sensor is capable of measuring the viscosity of liquid phase samples with a resolution of 0.15 cP by tracking the Q-factor response of the sensor within a linear regime between 1 to 2.6 cP. This approach demonstrates the ability to identify in real-time changes of fluids in the liquid phase that could provide a valuable assessment for bioanalytical applications

looking .forward - The IEEE Computer Society's newsletter - A supplement to IEEE Computer - Spring 1998 - Vol 6, nº 1

looking .forward - The IEEE Computer Society's newsletter - A supplement to IEEE Computer - Spring 1998 - Vol 6, nº 1Preprin

Metallic nanostructures based on blu-ray discs for multiplexed plasmonic biodetection

Resumen del trabajo presentado al VIII International Congress on Analytical Nanoscience and Nanotechnology, celebrado en Barcelona (España) del 3 al 5 de julio de 2017.Nanostructure-based plasmonic biosensors have quickly positioned themselves as interesting candidates for the design of portable optical biosensor platforms considering the potential benefits they can offer in miniaturization, multiplexing, real-time and label-free detection. We have developed a simple integrated nanoplasmonic sensor taking advantage of the periodic nanostructured array of commercial Blu-ray discs. Sensors with different metallic film layers (50 and 100 nm) were fabricated and optically characterized by varying the oblique-angle of the incident light in optical reflectance measurements. We observed an enhancement in sensitivity and a narrowing of the resonant linewidths as the light incidence angle was increased, which could be related to the generation of Fano resonant modes. The new sensors achieve a figure of merit (FOM) up to 35 RIU-1 and a competitive bulk limit of detection (LOD) of 6.34×10-6 RIU. Based on these structures, a single-channel sensor in a 1cm2 integrated chip has been designed an incorporated in a compact biosensor prototype. The label-free biosensing capability of the new sensor has been assessed by evaluating the presence of specific antibodies against the GTF2b protein, a tumor-associate antigen (TAA) related to colorectal cancer. We have achieved a LOD in the pM order and have evaluated the feasibility of directly measuring biological samples such as human serum. The design has been transferred and adapted to a multiplexed prototype incorporating a sensor chip with four integrated channels. The optical and the sensing performance have been evaluated. We have assessed bulk sensitivity and attempted real-time biosensing measurements, obtaining good reproducibility between the channels.Peer reviewe