Oriented Tailoring of Plastic Antibodies for Prostate Specific Antigen and Application of the Imprinted Material as Ionophore in Potentiometric Detection

Poster, presented at Nanobio Europe, Varese, Italy, 18 - 20 June, 2012.Prostate Specific Antigen (PSA) is the biomarker of choice for screening prostate cancer throughout the population, with PSA values above 10 ng/mL pointing out a high probability of associated cancer1. According to the most recent World Health Organization (WHO) data, prostate cancer is the commonest form of cancer in men in Europe2. Early detection of prostate cancer is thus very important and is currently made by screening PSA in men over 45 years old, combined with other alterations in serum and urine parameters. PSA is a glycoprotein with a molecular mass of approximately 32 kDa consisting of one polypeptide chain, which is produced by the secretory epithelium of human prostate. Currently, the standard methods available for PSA screening are immunoassays like Enzyme-Linked Immunoabsorbent Assay (ELISA). These methods are highly sensitive and specific for the detection of PSA, but they require expensive laboratory facilities and high qualify personal resources. Other highly sensitive and specific methods for the detection of PSA have also become available and are in its majority immunobiosensors1,3-5, relying on antibodies. Less expensive methods producing quicker responses are thus needed, which may be achieved by synthesizing artificial antibodies by means of molecular imprinting techniques. These should also be coupled to simple and low cost devices, such as those of the potentiometric kind, one approach that has been proven successful6. Potentiometric sensors offer the advantage of selectivity and portability for use in point-of-care and have been widely recognized as potential analytical tools in this field. The inherent method is simple, precise, accurate and inexpensive regarding reagent consumption and equipment involved. Thus, this work proposes a new plastic antibody for PSA, designed over the surface of graphene layers extracted from graphite. Charged monomers were used to enable an oriented tailoring of the PSA rebinding sites. Uncharged monomers were used as control. These materials were used as ionophores in conventional solid-contact graphite electrodes. The obtained results showed that the imprinted materials displayed a selective response to PSA. The electrodes with charged monomers showed a more stable and sensitive response, with an average slope of -44.2 mV/decade and a detection limit of 5.8X10-11 mol/L (2 ng/mL). The corresponding non-imprinted sensors showed smaller sensitivity, with average slopes of -24.8 mV/decade. The best sensors were successfully applied to the analysis of serum samples, with percentage recoveries of 106.5% and relatives errors of 6.5%

Automated in situ consolidation process for pre-impregnated carbon fibers: a cyber phisycal approach

The concept known as the fourth industrial revolution (Industry 4.0) has developed a wide field of study, such as the Internet of Things (IoT), Cyber-Physical Systems (CPS) and Information and Communication Technology (ICT), with the objective to achieve enhancing the processes performance of automated systems. This work focuses on the field of CPS for an in situ consolidation machine, which handles preimpregnated, in this case unidirectional carbon fiber tapes with polyamide 6, in order to create composite laminates. The physical model involves an unwinding system that feeds the tape into a turning table through a pressing mechanism, which has a radiation heater before and refrigeration after the presser to guarantee the composite temperature. The objective is to model and simulate the heating process before, during and after the pressing mechanism to feed an optimization algorithm in order to calculate the optimal working parameters such as heaters power, pressing force and process speed taking into account the temperatures of the tape, to avoid overheating and to improve process time. The results are processed, so that they can send and receive data constantly, to an optimization system to alter parameters, without making programing code changes and avoiding the need to stop the consolidation. The CPS can handle different classes of pre-impregnated composites, by learning during the consolidation process, in order to improve the overall result, without the need of fully or partially reprograming the code, which a main characteristic of CPS

Green approaches for extraction, chemical modification and processing of marine polysaccharides for biomedical applications

Over the past few decades, natural-origin polysaccharides have received increasing attention across different fields of application, including biomedicine and biotechnology, because of their specific physicochemical and biological properties that have afforded the fabrication of a plethora of multifunctional devices for healthcare applications. More recently, marine raw materials from fisheries and aquaculture have emerged as a highly sustainable approach to convert marine biomass into added-value polysaccharides for human benefit. Nowadays, significant efforts have been made to combine such circular bio-based approach with cost-effective and environmentally-friendly technologies that enable the isolation of marine-origin polysaccharides up to the final construction of a biomedical device, thus developing an entirely sustainable pipeline. In this regard, the present review intends to provide an up-to-date outlook on the current green extraction methodologies of marine-origin polysaccharides and their molecular engineering toolbox for designing a multitude of biomaterial platforms for healthcare. Furthermore, we discuss how to foster circular bio-based approaches to pursue the further development of added-value biomedical devices, while preserving the marine ecosystem.publishe

Bioinspired oxidation-resistant catechol-like sliding ring polyrotaxane hydrogels

Adaptable hydrogels have been used in the biomedical field to address several pathologies, especially those regarding tissue defects. Here, we describe unprecedented catechol-like functionalized polyrotaxane (PR) polymers able to form hydrogels. PR were functionalized with the incorporation of hydroxypyridinone (HOPO) moieties into the polymer backbone, with a degree of substitution from 4 to 22%, depending on the PR type. The hydrogels form through the functionalized supramolecular systems when in contact with a Fe(III) solution. Despite the hydrogel formation being at physiological pH (7.4), the HOPO derivatives are extremely resistant to oxidation, unlike common catechols; consequently, they prevent the formation of quinones, which can lead to irreversible bounds within the matrix. The resulting hydrogels demonstrated properties lead to unique hydrogels with improved mechanical behavior obtained by metallic coordination crosslinking, due to the synergies of the sliding-ring PR and the non-covalent (reversible) catechol analogues. Following this strategy, we successfully developed innovative, cytocompatible, oxidative-resistant, and reversible crosslinked hydrogels, with the potential of being used as structural self-materials for a variety of applications, including in the biomedical field.publishe

Novel Prostate Specific Antigen plastic antibody designed withcharged binding sites for an improved protein binding and itsapplication in a biosensor of potentiometric transduction

This work shows that the synthesis of protein plastic antibodies tailored with selected charged monomersaround the binding site enhances protein binding. These charged receptor sites are placed over a neutralpolymeric matrix, thus inducing a suitable orientation the protein reception to its site. This is confirmed bypreparing control materials with neutral monomers and also with non-imprinted template. This concepthas been applied here to Prostate Specific Antigen (PSA), the protein of choice for screening prostate can-cer throughout the population, with serum levels >10 ng/mL pointing out a high probability of associatedcancer.Protein Imprinted Materials with charged binding sites (C/PIM) have been produced by surfaceimprinting over graphene layers to which the protein was first covalently attached. Vinylben-zyl(trimethylammonium chloride) and vinyl benzoate were introduced as charged monomers labellingthe binding site and were allowed to self-organize around the protein. The subsequent polymerizationwas made by radical polymerization of vinylbenzene. Neutral PIM (N/PIM) prepared without orientedcharges and non imprinted materials (NIM) obtained without template were used as controls.These materials were used to develop simple and inexpensive potentiometric sensor for PSA. Theywere included as ionophores in plasticized PVC membranes, and tested over electrodes of solid or liq-uid conductive contacts, made of conductive carbon over a syringe or of inner reference solution overmicropipette tips. The electrodes with charged monomers showed a more stable and sensitive response,with an average slope of -44.2 mV/decade and a detection limit of 5.8 × 10−11mol/L (2 ng/mL). The cor-responding non-imprinted sensors showed lower sensitivity, with average slopes of -24.8 mV/decade.The best sensors were successfully applied to the analysis of serum, with recoveries ranging from 96.9to 106.1% and relative errors of 6.8%

Betão fibroso de comportamento melhorado ao fogo

As Universidades do Minho e de Coimbra estão a desenvolver um betão reforçado com fibras (BRF), de elevada ductilidade, com comportamento melhorado sob a acção de temperaturas elevadas. Este betão inclui fibras de polipropileno e fibras de aço. Nesta comunicação, apresentam-se os resultados relativos à avaliação da influência de temperaturas elevadas nas seguintes propriedades mecânicas residuais do betão fibroso desenvolvido: módulo de elasticidade, resistência à compressão e resistência à flexão. A aplicabilidade de ensaios não destrutivos na avaliação das propriedades residuais do betão é explorada.Os autores pretendem agradecer o apoio disponibilizado pela FCT (Projecto FICOFIRE - POCTI/ECM/57518/2004), Spie Batignolles, Civitest, Secil, Sika, DuroEuropa e Miguel Freitas. Os quarto e quinto autores agradecem o apoio financeiro obtido por intermédio das respectivas bolsas, ao abrigo do projecto acima referido

Realtime local navigation for the blind: detection of lateral doors and sound interface

Worldwide there are about 285 million visually impaired persons, of which 39 million are blind and the others have low vision. Almost all systems designed to assist them are quite complex and expensive, but most blind persons do not have advanced technical assistance and they are rather poor. We are therefore developing a low-cost navigation aid which can be afforded by almost all blind persons: basically, the ultimate goal is to use only a mobile phone with a built-in camera. This aid complements the white cane, it is easily portable, and it is not a hindrance when walking with the cane. The system will have an easy and intuitive interface, yet providing assistance in local and global navigation in realtime. In this paper we present the progress concerning local navigation. Path and obstacle detection just beyond the reach of the cane is now supplemented by detection of doors in corridors. This is necessary for localization, i.e., for developing a better impression of the environment and for finding a specific room. A sophisticated sound interface can assist the user for centering on paths like sidewalks and corridors, alerting to looming obstacles for avoiding them

Sarcosine oxidase composite screen-printed electrode for sarcosine determination in biological samples

XIX Meeting of the Portuguese Electrochemical Society - XVI Iberic Meeting of ElectrochemistryProstate Cancer (PCa) is the most common form of cancer in men in Europe with a 61.4 % incidence among all cancer cases and a 12.1 % mortality [1] and, therefore, its early detection is fundamental for increasing the survival rate. Currently, diagnosis and management of patients with PCa is only based on the determination of the biomarker Prostate Specific Antigen (PSA). However, the method used for PCa detection has poor sensitivity and specificity, leading to false negative and false positive test results and many patients are sent to unnecessary biopsy procedures [2]. Therefore, there is a need to seek for new biomarkers and more effective screening. In this work, a biosensor device was developed for the quantification of sarcosine via electrochemical detection of H2O2 (at 0.6 V) generated from the catalyzed oxidation of sarcosine. The detection was carried out after the modification of carbon screen printed electrodes (SPEs) by immobilization of sarcosine oxidase (SOX) on the electrode surface. The strategies used herein included the activation of the carbon films by an electrochemical step and the formation of an NHS/EDAC layer to bond the enzyme to the electrode, the use of metallic or semiconductor nanoparticles layer previously or during the enzyme immobilization. In order to improve the sensor stability and selectivity a polymeric layer with extra enzyme content was further added. The proposed methodology for the detection of sarcosine allowed obtaining a limit of detection (LOD) of 1.6x10-5 mM, using a linear concentration range between 1x10-5 and 1x10-4 mM. The biosensor was successfully applied to the analysis of sarcosine in urine samples.

Kidney targeting and puncturing during percutaneous nephrolithotomy: recent advances and future perspectives

Background and Purpose: Precise needle puncture of the kidney is a challenging and essential step for successful percutaneous nephrolithotomy (PCNL). Many devices and surgical techniques have been developed to easily achieve suitable renal access. This article presents a critical review to address the methodologies and techniques for conducting kidney targeting and the puncture step during PCNL. Based on this study, research paths are also provided for PCNL procedure improvement. METHODS: Most relevant works concerning PCNL puncture were identified by a search of Medline/PubMed, ISI Web of Science, and Scopus databases from 2007 to December 2012. Two authors independently reviewed the studies. RESULTS: A total of 911 abstracts and 346 full-text articles were assessed and discussed; 52 were included in this review as a summary of the main contributions to kidney targeting and puncturing. CONCLUSIONS: Multiple paths and technologic advances have been proposed in the field of urology and minimally invasive surgery to improve PCNL puncture. The most relevant contributions, however, have been provided by the application of medical imaging guidance, new surgical tools, motion tracking systems, robotics, and image processing and computer graphics. Despite the multiple research paths for PCNL puncture guidance, no widely acceptable solution has yet been reached, and it remains an active and challenging research field. Future developments should focus on real-time methods, robust and accurate algorithms, and radiation free imaging techniques.The authors acknowledge Foundation for Science and Technology (FCT) for the fellowships references: SFRH/BPD/46851/2008 and SFRH/BD/74276/2010