Development of an impedance based biosensor for studies of spheroids formation

Three-dimensional (3D) multicellular spheroids are exceptional in vitro cell models for their ability to accurately mimic real cell-cell interaction processes. However, the challenges in producing well-defined spheroids with controlled size together with the deficiency of techniques to monitor them significantly restrict their use. Herein, a novel device to study spheroid formation in real time is presented. By exploiting electrochemical impedance spectroscopy, a multi-electrode array (MEA) attached to a calcium alginate scaffold is able to monitor the behaviour of 36 different hydrogel wells. The scaffold contains inverted shape pyramidal microwells, which guide the aggregation of cells into spheroids with controlled dimensions. Preliminar studies on calcium alginate, optimisation of fabrication strategy are shown, together with testing of the device in the presence and the absence of the hydrogel. Lastly, the device was tested for its intended aim, i.e. to monitor the formation of a spheroid, proving its potential as an impedance biosensor.

Development of wearable electrochemical sensors for the determination of biological analytes

Wearable biosensors are attracting interest due to their potential to provide continuous, real-time physiological information via dynamic, non-invasive measurements of biochemical markers in biofluids, such as interstitial fluid (ISF). One notable example of their applications is for glycemic monitoring in diabetic patients, which is typically carried out either by direct measurement of blood glucose via finger pricking or by wearable sensors that can continuously monitor glucose in ISF by sampling it from below the skin with a microneedle. In this context, the development of a new and minimally invasive multisensing tattoo-based platform for the monitoring of glucose and other analytes in ISF extracted through reverse iontophoresis in proposed by the GLUCOMFORT project. This elaborate describes the in-vitro development of flexible electrochemical sensors based on inkjet-printed PEDOT:PSS and metal inks that are capable of determining glucose and chloride at biologically relevant concentrations, making them good candidates for application in the GLUCOMFORT platform. In order to make PEDOT:PSS sensitive to glucose at micromolar concentrations, a biocompatible functionalization based on immobilized glucose oxidase and electrodeposited platinum was developed. This functionalization was successfully applied to bulk and flexible amperometric devices, the design of which was also optimized. Using the same strategy, flexible organic electrochemical transistors (OECTs) for glucose sensing were also made and successfully tested. For the sensing of chloride ions, an organic charge-modulated field-effect transistor (OCMFET) featuring a silver/silver chloride modified floating gate electrode was developed and tested.

Sviluppo di biosensori reagentless basati sull'enzima fosfatasi alcalina

Negli ultimi anni i biosensori enzimatici sono diventati sempre più popolari nel campo delle determinazioni rapide di analiti di interesse industriale ed ambientale. L’utilizzo di tali biosensori consente di evitare, o comunque limitare, l’impiego di metodi analitici basati su tecniche più complesse e dispendiose in termini economici e temporali. Un grande obbiettivo della ricerca negli ultimi anni è quello di costruire biosensori reagentless, ossia dispositivi pronti all’uso da parte degli analisti, indipendentemente dalla loro esperienza. In questo lavoro di tesi sono stati sviluppati dei biosensori amperometrici reagentless che sfruttano l’enzima fosfatasi alcalina (ALP), come elemento di riconoscimento biologico, ed un elettrodo screen-printed (SPE) commerciale, come trasduttore. Per gli elettrodi SPE commerciali sono stati testati modificanti a base di nanomateriali carboniosi e del polimero conduttore PEDOT:ClO4. Sono inoltre stati messi a punto diversi biosensori costruiti utilizzando sia l’enzima ALP immobilizzato che lo stesso enzima in soluzione. I test sono stati eseguiti utilizzando il substrato enzimatico “ascorbil-fosfato di sodio (AAP)” ed alcuni inibitori enzimatici. I dispositivi reagentless sono stati fabbricati mediante stampa 3D e sono stati realizzati appositamente per gli elettrodi SPE commerciali utilizzati. Tali dispositivi sono stati utilizzati per effettuare la determinazione dell’AAP in campo cosmetico e la determinazione dell’enzima ALP in campo alimentare.

As provas de matemática no acesso à universidade : um estudo sobre o processo seletivo seriado (PSS) na Universidade Federal da Paraíba

Este trabalho voltou-se para o processo de seleção para ingresso nas Universidades Brasileiras, em particular o realizado pela Universidade Federal da Paraíba, a qual utiliza duas formas: vestibular tradicional e Processo Seletivo Seriado (PSS). O PSS, em particular as provas de Matemática, constitui centro de interesse da pesquisa cujo objetivo é investigar a opinião de professores a respeito de conteúdo contemplado, grau de dificuldade exigido e aplicabilidade dos Parâmetros Curriculares Nacionais nas provas. O presente estudo caracteriza-se como uma pesquisa exploratória, descritiva. Tendo como sujeitos professores de Matemática das escolas públicas e privadas que oferecem o curso médio, localizadas na cidade de João Pessoa. Os resultados apontam que não foram observadas as diretrizes Curriculares Nacionais no que tange a conteúdos, habilidades e competências que servem de referência para avaliação do desempenho dos alunos do Ensino Médio nas soluções de problemas matemáticos com diferentes graus de dificuldade. Consideramos que os conteúdos privilegiados continuam sendo os determinados pela comissão do vestibular em seu documento norteador o manual do candidato.

Prato e Sabadell: quali prospettive per due "città del tessile"?

Prato e Sabadell hanno avuto peculiarità territoriali ed economiche molto simili, tanto da essere state definite nel tempo come due “città del tessile”. Entrambe le città, infatti, hanno avuto una tradizione manifatturiera tessile con una simile organizzazione della attività produttiva. Da un punto di vista politico, questo lavoro apporta preziosi elementi per un studio paragonativo tra le “due città del tessile”.

PSS Development and Training Strategy 2006-2016 (PDF 149KB)

This Circular details arrangements for securing the strategic priorities of the PSS Development and Training Strategy (the Strategy) in Northern Ireland from 1 April 2012 onwards. åÊThis replaces the previous circular HSS (OSS) Training 1/2010.

Key Indicators of PSS for NI (2002)

Key Indicators of PSS for NI (2002)

Suomen maan pedot maalikuwilla selitetyt - [Kuvaliite]

Turku : J.W. Lillja & Co. 1854

"Askeleen päässä taidosta" - Kolmen askeleen menetelmän soveltuvuuden testaaminen oppimistehtävän avulla motoristen taitojen opetukseen 4. luokan käsityössä

Tutkimuksen tarkoituksena on selvittää taitojen opettamisen kolmen askeleen menetelmän soveltuvuutta motoristen taitojen opetukseen käsityön 4. luokalla. Menetelmän soveltuvuuden testaamiseksi on suunniteltu ja valmistettu oppimistehtävä. Oppimistehtävä on laadittu taitojen opettamisen yleismenetelmän, kolmen askeleen menetelmän pohjalta. Oppimistehtävän laadinnassa on käytetty tukena myös suljetun oppimistehtävän teoreettista perustaa, sekä ositetun ja taitolajipainotteisen käsityön piirteitä. Oppimistehtävä toteutettiin satakuntalaisessa koulussa ja toteutuksen suoritti koulun oma käsityönopettaja. Oppimistehtävän aikana tapahtuvasta toiminnasta, opetuksesta ja oppimisessa on pyritty saamaan tietoa kaikilta siihen osallistuneilta osapuolilta. Oppilaille on suoritettu oppimista mittaava alku ja loppu kysely. Opettajaa on haastateltu, jolloin on saatu oppimistehtävän toteuttajan kokemuksia tapahtuneesta. Lisäksi tutkijat ovat itse osallistuneet toiminnan havainnointiin, sekä videoineet opetusta ja oppilaiden toimintaa. Tutkimus on tutkimusotteeltaan laadullinen tapaustutkimus. Tutkimuksen tuote on motoristen taitojen opettamiseen laadittu oppimistehtävä. Tutkimuksen kulku ja toteutus on kuvattu yksityiskohtaisesti mittarien laadinnasta, aineiston keruun ja -tulosten analysoinnin kautta synteesiin ja tutkimusongelmiin vastaamiseen. Tutkimuksen tulokset osoittavat, että taitojen opettamisen kolmen askeleen menetelmä sisältää myös perusopetuksessa toteutettavan taitojen opetuksen kannalta keskeisimmät elementit. Menetelmä soveltuu etenkin vasta-alkaneille opettajille, mutta myös kokeneille opettajille. Taitojen oppimisen kannalta erityisen tärkeäksi seikaksi tällä ikäryhmällä osoittautui demonstrointi ja ohjaaminen. Lisäksi saimme yksityiskohtaista tietoa, jonka avulla menetelmästä voisi laatia tälle ikäryhmälle vieläkin sopivamman.

Solid-state reference and ion-selective electrodes : towards portable potentiometric sensing

Potentiometric sensors are very attractive tools for chemical analysis because of their simplicity, low power consumption and low cost. They are extensively used in clinical diagnostics and in environmental monitoring. Modern applications of both fields require improvements in the conventional construction and in the performance of the potentiometric sensors, as the trends are towards portable, on-site diagnostics and autonomous sensing in remote locations. The aim of this PhD work was to improve some of the sensor properties that currently hamper the implementation of the potentiometric sensors in modern applications. The first part of the work was concentrated on the development of a solid-state reference electrode (RE) compatible with already existing solid-contact ion-selective electrodes (ISE), both of which are needed for all-solid-state potentiometric sensing systems. A poly(vinyl chloride) membrane doped with a moderately lipophilic salt, tetrabutylammonium-tetrabutylborate (TBA-TBB), was found to show a satisfactory stability of potential in sample solutions with different concentrations. Its response time was nevertheless slow, as it required several minutes to reach the equilibrium. The TBA-TBB membrane RE worked well together with solid-state ISEs in several different situations and on different substrates enabling a miniature design. Solid contacts (SC) that mediate the ion-to-electron transduction are crucial components of well-functioning potentiometric sensors. This transduction process converting the ionic conduction of an ion-selective membrane to the electronic conduction in the circuit was studied with the help of electrochemical impedance spectroscopy (EIS). The solid contacts studied were (i) the conducting polymer (CP) poly(3,4-ethylienedioxythiophene) (PEDOT) and (ii) a carbon cloth having a high surface area. The PEDOT films were doped with a large immobile anion poly(styrene sulfonate) (PSS-) or with a small mobile anion Cl-. As could be expected, the studied PEDOT solid-contact mediated the ion-toelectron transduction more efficiently than the bare glassy carbon substrate, onto which they were electropolymerized, while the impedance of the PEDOT films depended on the mobility of the doping ion and on the ions in the electrolyte. The carbon cloth was found to be an even more effective ion-to-electron transducer than the PEDOT films and it also proved to work as a combined electrical conductor and solid contact when covered with an ion-selective membrane or with a TBA-TBB-based reference membrane. The last part of the work was focused on improving the reproducibility and the potential stability of the SC-ISEs, a problem that culminates to the stability of the standard potential E°. It was proven that the E° of a SC-ISE with a conducting polymer as a solid contact could be adjusted by reducing or oxidizing the CP solid contact by applying current pulses or a potential to it, as the redox state of the CP solid-contact influences the overall potential of the ISE. The slope and thus the analytical performance of the SC-ISEs were retained despite the adjustment of the E°. The shortcircuiting of the SC-ISE with a conventional large-capacitance RE was found to be a feasible instrument-free method to control the E°. With this method, the driving force for the oxidation/reduction of the CP was the potential difference between the RE and the SC-ISE, and the position of the adjusted potential could be controlled by choosing a suitable concentration for the short-circuiting electrolyte. The piece-to-piece reproducibility of the adjusted potential was promising, and the day-today reproducibility for a specific sensor was excellent. The instrumentfree approach to control the E° is very attractive considering practical applications.