567 research outputs found
Nonequilibrium Thermodynamics And Quasielastic Light Scattering From Crystals. Ii. Piezoelectric Crystals
The methodology described and utilized previously by the author, involving the use of nonequilibrium thermodynamics in the analysis of the spectrum of light scattered quasielastically by crystals, is applied to piezoelectric crystals. An additional thermodynamic variable, the electric polarization, is taken into account and assumed to be a relaxing variable. Solution of the resulting equations in the case of KH2PO4 leads, as expected, to the appearance of a polarization relaxation line in the spectrum. The spectrum is calculated for several values of the two relevant parameters: the polarization relaxation frequency and the ratio of C66 isothermal elastic constants at constant electric field and constant polarization. For a fixed polarization-relaxation frequency, it is found that as the ratio of elastic constants decreases (the characteristic behavior as the ferroelectric phase transition in KH2PO4 is approached), the intensity of the relaxation line increases at the expense of the Brillouin lines. In contrast with standard treatments of coupled modes in piezoelectric crystals, no adjustable parameters are involved in determining the spectrum. © 1980 The American Physical Society.22126006601
SYNTHESIS OF RGD PEPTIDOMIMETIC-DRUG CONJUGATES FOR THE TUMOR-TARGETED DELIVERY OF CYTOTOXIC AGENTS
Integrins are a large family of heterodimeric transmembrane glycoprotein receptors, composed by two non-covalently associated subunits (\u3b1 and \u3b2). Integrins \u3b1V\u3b23 and \u3b1V\u3b25 are overexpressed on blood vessels in human tumors but not on vessels in normal tissues and, for this reason, they have become attractive targets for pharmacological studies in oncology. Thus, in this PhD thesis, the synthesis of new Small Molecule-Drug Conjugates (SMDCs) targeting \u3b1v\u3b23 integrin is described. The structure of such SMDCs consists in the connection of three core components: i) LIGAND - the cyclo[DKP-RGD] peptidomimetic, developed by the Gennari and Piarulli group, has been used as integrin-targeting moiety in all the new constructs; ii) CYTOTOXIC PAYLOAD - three different cytotoxic agents (i.e. paclitaxel, monomethyl auristatin E and monomethyl auristatin F) have been included as anticancer drugs; iii) LINKER - specific functional groups (i.e. peptides) have been used to connect the drug and ligand, aiming at the selective drug release in the intra or extracellular tumor environment. These new SMDCs have been subjected to a panel of biochemical and biological assays, for assessing both their structural features (e.g. stability, kinetics of drug release, etc.) and biological activity (e.g. affinity for the purified integrin \u3b1V\u3b23 receptor, selective cytotoxicity against \u3b1v\u3b23-expressing or non-expressing cells, etc.).
As a first project, Chapter II describes the development of a new library of multimeric cyclo[DKP-RGD]-PTX conjugates (monomeric, dimeric, trimeric and tetrameric conjugates) bearing lysosomally-cleavable linkers and their full in vitro biological evaluation. The results achieved with these first compounds prompted the design of next-generation cyclo[DKP-RGD]-PTX conjugates, reported in the following chapters. Chapter III describes the synthesis and in vitro evaluation of cyclo[DKP-RGD]-PTX conjugates bearing extracellularly-cleavable peptide linkers, capable of releasing the payload in the tumor microenvironment, rather than inside cancer cells: this mechanism can be promoted by tumor-associated enzymes present in the tumor stroma (e.g. elastase), which can efficiently cleave the linker and set the drug free to diffuse within the tumor mass. Finally, Chapter IV consists in the development of a small library of conjugates containing the cyclo[DKP-RGD] ligand, a lysosomally-cleavable peptide linker and the highly potent toxins monomethyl auristatin E or F (MMAE and MMAF) as the cytotoxic payloads, which are state-of-the-art tools for targeted anticancer therapy
Myoglobin-biomimetic electroactive materials made by surface molecular imprinting on silica beads and their use as ionophores in polymeric membranes for potentiometric transduction
Myoglobin (Mb) is among the cardiac biomarkers playing a major role in urgent diagnosis of cardiovascular diseases. Its monitoring in point-of-care is therefore fundamental. Pursuing this goal, a novel biomimetic ionophore for the potentiometric transduction of Mb is presented. It was synthesized by surface molecular imprinting (SMI) with the purpose of developing highly efficient sensor layers for near-stereochemical recognition of Mb. The template (Mb) was imprinted on a silane surface that was covalently attached to silica beads by means of self-assembled monolayers. First the silica was modified with an external layer of aldehyde groups. Then, Mb was attached by reaction with its amine groups (on the external surface) and subsequent formation of imine bonds. The vacant places surrounding Mb were filled by polymerization of the silane monomers 3-aminopropyltrimethoxysilane (APTMS) and propyltrimethoxysilane (PTMS). Finally, the template was removed by imine cleavage after treatment with oxalic acid. The results materials were finely dispersed in plasticized PVC selective membranes and used as ionophores in potentiometric transduction. The best analytical features were found in HEPES buffer of pH 4. Under this condition, the limits of detection were of 1.3 × 10−6 mol/L for a linear response after 8.0 × 10−7 mol/L with an anionic slope of −65.9 mV/decade. The imprinting effect was tested by preparing non-imprinted (NI) particles and employing these materials as ionophores. The resulting membranes showed no ability to detect Mb. Good selectivity was observed towards creatinine, sacarose, fructose, galactose, sodium glutamate, and alanine. The analytical application was conducted successfully and showed accurate and precise results
Surface Imprinting Approach on Screen Printed Electrodes Coated with Carboxylated PVC for Myoglobin detection with Electrochemical Transduction
A novel surface molecularly-imprinted (MI) material to detect myoglobin (Myo) using gold screen printed electrodes (SPE) was developed. The sensitive detection was carry out by introducing a carboxylic polyvinyl chloride (PVC-COOH) layer on gold SPE surface. Myo was attached to the surface of gold SPE/PVC-COOH and the vacant spaces around it were filled by polymerizing acrylamide and N,N-methylenebisacrylamide (cross-linker). This polymerization was initiated by ammonium persulphate. After removing the template, the obtained material was able to rebind Myo and discriminate it among other interfering species. Various characterization techniques including electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) confirmed the surface modification. This sensor seemed a promising tool for screening Myo in point-of-care
Estudo da Sensibilização aos Aeroalergenos Phl p 1, Phl p 5, Ole e 1 e Ole e 2 em Doentes com Patologia Alérgica Sazonal
Introdução: A sensibilização aos pólenes depende de vários factores nomeadamente do tipo de vegetação local e sabe-se que a sintomatologia não está apenas associada à exposição aos pólens mas também a partÃculas, algumas das quais resultantes da rotura dos grãos de pólen sendo posteriormente aerossolizadas.
Objectivos: Relacionar a sensibilização de doentes com a concentração polÃnica atmosférica e a concentração de alguns dos respectivos aerolergenos.
Métodos: Das consultas externas de Imunoalergologia dos hospitais de Évora e Elvas seleccionaram-se doentes que apresentavam queixas sazonais de rinite alérgica e asma brônquica, aos quais foram realizados testes cutâneos em Prick, standardizados, aos pólenes identificados na região. A 55 doentes foram realizados testes ao extracto de Phleum, aos seus alergénios Phl p 1 e Phl p 5, bem como aos extractos das restantes gramÃneas e a 47 doentes foram realizados testes ao extracto de Olea, aos seus alergénios Ole e 1 e Ole e 2. Monitorizaram-se diariamente as partÃculas polÃnicas e os aeroalergenos mediante 2 colectores especÃficos para cada tipo.
Resultados: A percentagem de doentes que é sensÃvel aos 3 extractos de Phleum (Phleum total, Phl p 1 e Phl p 5) é de 51% , a dos que são sensÃveis aos extractos de Phleum total e Phl p 1 é de 16 % e a dos que são sensÃveis a Phleum total e Phl p5 é de 2%. A percentagem de doentes que é sensÃvel a somente um dos extractos é de 20% e os que não têm qualquer sensibilidade são 11%. A percentagem de doentes que é sensÃvel aos 3 extractos de Olea (Olea total, Ole e 1 e Ole e 2) é de 23%, a dos que são sensÃveis à Olea total e Ole e 1 é de 21 % e a dos que são sensÃveis à Olea total e Ole e 2 é de apenas 4%. A percentagem dos que são sensÃveis a um dos extractos é de 19% e a dos que não apresentaram qualquer sensibilidade é de 32%.
Conclusões: Podemos concluir que 89% dos doentes mostraram ser sensÃveis aos alergénios da gramÃnea Phleum pratense pois mostraram positividade tanto ao extracto de Phleum e/ou aos seus alergénios Phl p 1 e Phl p 5. Em relação à oliveira, 44% dos doentes são alérgicos a este pólen pois são sensÃveis ao seu alergénio major, Ole e 1. Em ambos os casos estão correlacionados com os aeroalergenos detectados nas amostras de ar, sendo a sensibilidade aos pólenes de gramÃneas maior que a sensibilidade ao pólen de oliveira. Mais estudos devem ser realizados para despiste de reacções cruzadas nomeadamente quanto à Olea com outras plantas da região
Artificial antibodies for troponin T by its imprinting on the surface of multiwalled carbon nanotubes: Its use as sensory surfaces
A novel artificial antibody for troponin T (TnT) was synthesized by molecular imprint (MI) on the surface of multiwalled carbon nanotubes (MWCNT). This was done by attaching TnT to the MWCNT surface, and filling the vacant spaces by polymerizing under mild conditions acrylamide (monomer) in N,N′-methylenebisacrylamide (cross-linker) and ammonium persulphate (initiator). After removing the template, the obtained biomaterial was able to rebind TnT and discriminate it among other interfering species. Stereochemical recognition of TnT was confirmed by the non-rebinding ability displayed by non-imprinted (NI) materials, obtained by imprinting without a template. SEM and FTIR analysis confirmed the surface modification of the MWCNT. The ability of this biomaterial to rebind TnT was confirmed by including it as electroactive compound in a PVC/plasticizer mixture coating a wire of silver, gold or titanium. Anionic slopes of 50 mV decade−1 were obtained for the gold wire coated with MI-based membranes dipped in HEPES buffer of pH 7. The limit of detection was 0.16 μg mL−1. Neither the NI-MWCNT nor the MWCNT showed the ability to recognize the template. Good selectivity was observed against creatinine, sucrose, fructose, myoglobin, sodium glutamate, thiamine and urea. The sensor was tested successfully on serum samples. It is expected that this work opens new horizons on the design of new artificial antibodies for complex protein structures
Novel biosensing device for point-of-care applications with plastic antibodies grown on Au-Screen Printed Electrodes
A gold screen printed electrode (Au-SPE) was modified by merging Molecular Imprinting and Self-Assembly Monolayer techniques for fast screening cardiac biomarkers in point-of-care (POC). For this purpose, Myoglobin (Myo) was selected as target analyte and its plastic antibody imprinted over a glutaraldehyde (Glu)/cysteamine (Cys) layer on the gold-surface. The imprinting effect was produced by growing a reticulated polymer of acrylamide (AAM) and N,N′-methylenebisacrylamide (NNMBA) around the Myo template, covalently attached to the biosensing surface. Electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) studies were carried out in all chemical modification steps to confirm the surface changes in the Au-SPE.
The analytical features of the resulting biosensor were studied by different electrochemical techniques, including EIS, square wave voltammetry (SWV) and potentiometry. The limits of detection ranged from 0.13 to 8 μg/mL. Only potentiometry assays showed limits of detection including the cut-off Myo levels. Quantitative information was also produced for Myo concentrations ≥0.2 μg/mL. The linear response of the biosensing device showed an anionic slope of ~70 mV per decade molar concentration up to 0.3 μg/mL. The interference of coexisting species was tested and good selectivity was observed. The biosensor was successfully applied to biological fluids
Protein-responsive polymers for point-of-care detection of cardiac biomarker
This work describes a novel use for the polymeric film, poly(o-aminophenol) (PAP) that was made responsive to a specific protein. This was achieved through templated electropolymerization of aminophenol (AP) in the presence of protein. The procedure involved adsorbing protein on the electrode surface and thereafter electroploymerizing the aminophenol. Proteins embedded at the outer surface of the polymeric film were digested by proteinase K and then washed away thereby creating vacant sites. The capacity of the template film to specifically rebind protein was tested with myoglobin (Myo), a cardiac biomarker for ischemia. The films acted as biomimetic artificial antibodies and were produced on a gold (Au) screen printed electrode (SPE), as a step towards disposable sensors to enable point-of-care applications.
Raman spectroscopy was used to follow the surface modification of the Au-SPE. The ability of the material to rebind Myo was measured by electrochemical techniques, namely electrochemical impedance spectroscopy (EIS) and square wave voltammetry (SWV). The devices displayed linear responses to Myo in EIS and SWV assays down to 4.0 and 3.5 μg/mL, respectively, with detection limits of 1.5 and 0.8 μg/mL. Good selectivity was observed in the presence of troponin T (TnT) and creatine kinase (CKMB) in SWV assays, and accurate results were obtained in applications to spiked serum. The sensor described in this work is a potential tool for screening Myo in point-of-care due to the simplicity of fabrication, disposability, short time response, low cost, good sensitivity and selectivity
Smart Plastic Antibody Material (SPAM) tailored on disposable screen printed electrodes for protein recognition: application to Myoglobin detection
This work introduces two major changes to the conventional protocol for designing plastic antibodies: (i) the imprinted sites were created with charged monomers while the surrounding environment was tailored using neutral material; and (ii) the protein was removed from its imprinted site by means of a protease, aiming at preserving the polymeric network of the plastic antibody. To our knowledge, these approaches were never presented before and the resulting material was named here as smart plastic antibody material (SPAM).
As proof of concept, SPAM was tailored on top of disposable gold-screen printed electrodes (Au-SPE), following a bottom-up approach, for targeting myoglobin (Myo) in a point-of-care context. The existence of imprinted sites was checked by comparing a SPAM modified surface to a negative control, consisting of similar material where the template was omitted from the procedure and called non-imprinted materials (NIMs). All stages of the creation of the SPAM and NIM on the Au layer were followed by both electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV). AFM imaging was also performed to characterize the topography of the surface.
There are two major reasons supporting the fact that plastic antibodies were effectively designed by the above approach: (i) they were visualized for the first time by AFM, being present only in the SPAM network; and (ii) only the SPAM material was able to rebind to the target protein and produce a linear electrical response against EIS and square wave voltammetry (SWV) assays, with NIMs showing a similar-to-random behavior. The SPAM/Au-SPE devices displayed linear responses to Myo in EIS and SWV assays down to 3.5 μg/mL and 0.58 μg/mL, respectively, with detection limits of 1.5 and 0.28 μg/mL. SPAM materials also showed negligible interference from troponin T (TnT), bovine serum albumin (BSA) and urea under SWV assays, showing promising results for point-of-care applications when applied to spiked biological fluids
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