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1.
Biomater Biosyst ; 7: 100060, 2022 Aug.
Article in English | MEDLINE | ID: mdl-36824490

ABSTRACT

We describe the preparation of a membrane composed of polypyrrole-polystyrene (PPy-PS) and its application in DNA extraction. We adopted the electrospinning technique to prepare polystyrene (PS) membranes, which we used as substrates for incorporating polypyrrole chains through an in situ chemical procedure. As a model system, we initially investigated the use of PPy-PS membranes for the extraction of salmon sperm DNA from aqueous solutions. These studies have shown that the PPy-PS membrane has a maximum adsorption capacity of 236.0 mg of DNA per gram of PPy after 30 min of exposure to a DNA solution (100 mg/L). We incorporated the PPy-PS membranes into centrifugation columns, which we used to carry out experiments for extracting and purification of DNA from curly lettuce leaves. The protocol was initially optimized by first examining the most appropriate concentration of the three components of the lysis buffer (Tris/HCl, NaCl, and EDTA-Na). We then investigated the most adequate volumes of the concentrated surfactant solution (SDS 20%) and that used in the protein and polysaccharide precipitation step (5 M potassium acetate, pH 6.3), factors that directly influence the quality and quantity of the fraction of DNA obtained. For curly lettuce leaves, both in their mature and young stages, the yield and purity of the DNA purified using the PPy-PS membrane were comparable to those obtained using a commercial kit. In both cases, the collected DNA samples presented excellent integrity and quality. These results are suggestive that these composite membranes are competitive with the commercial kits available for the extraction and purification of DNA from plants.

2.
Colloids Surf B Biointerfaces ; 208: 112120, 2021 Dec.
Article in English | MEDLINE | ID: mdl-34597940

ABSTRACT

In 2016, the Zika virus (ZIKV) infection became a major public health problem, after the discovery that an alarming increase in the number of Brazilian newborns with microcephaly could be associated with the occurrence of this viral disease during the pregnancy of their mothers. The urgent need for simple diagnostic methods that allow rapid screening of suspected cases has stimulated the search for low-cost devices capable of detecting specific sequences of nucleic acids. The present work describes the development of nanostructured films formed by bilayers of conjugated polymers for rapid detection of the presence of Zika virus DNA, via fluorescence methods. For this, we initially deposited alternating layers of polyaniline (PANI) and polypyrrole (PPY) on the surface of polyethylene terephthalate (PET) sheets. The films obtained were then characterized by SEM, UV-Vis, ATR-FTIR, and contact angle measurements. For their use as quenchers for the diagnosis of Zika, a single DNA strand-specific for ZIKV was labeled with a fluorophore (FAM-ssDNA). We determined the time required for the saturation of the interaction between probe FAM-ssDNA and the film (180 min) and the time for the maximal hybridization between FAM-ssDNA and target DNA to occur (60 min). The detection limits were estimated as 345 pM and 278 pM for the PET/PPY-PANI and PET/PANI-PPY hybrid films, respectively. The simplicity of the procedure, coupled with the fact that a positive/negative response can be obtained in less than 60 min, suggests that the proposal of using these polymeric bilayer films is a promising methodology for the development of rapid molecular diagnostic tests.


Subject(s)
Pregnancy Complications, Infectious , Zika Virus Infection , Zika Virus , Electric Conductivity , Female , Humans , Infant, Newborn , Polymers , Pregnancy , Pregnancy Complications, Infectious/virology , Pyrroles , Zika Virus/genetics , Zika Virus Infection/diagnosis
3.
Anal Chim Acta ; 1178: 338762, 2021 Sep 15.
Article in English | MEDLINE | ID: mdl-34482873

ABSTRACT

We report the synthesis and characterization of a new hybrid magnetic composite formed by the enveloping of magnetic iron oxide nanoparticles (γ-NP) with chains of the conductive polymer PEDOT, and its use for the efficient separation of DNA molecules from complex biological samples, allowing the high yield separation of a pure and high-quality DNA fraction. The successful formation of the γ-NP/PEDOT composite was confirmed by Fourier transform infrared spectroscopy, scanning electron microscopy, UV visible spectroscopy (UV-Vis), and magnetic hysteresis loop measurements. The nanocomposites showed an excellent capacity of DNA adsorption (Qe âˆ¼ 248 mg/g) in a model system consisting of salmon sperm DNA. When the γ-NP/PEDOT was used in protocols to extract the DNA from complex samples, the corresponding yield was in the range of 6.4 µg (blood) and 7.3 µg (bacteria), as evaluated quality by UV-Vis, PCR analysis, and electrophoresis assays. We also established that the captured DNA does not need to be detached from the nanocomposite for use as seeding material in PCR amplification experiments. These results and the simplicity of the protocols indicate that the γ-NP/PEDOT composite is a promising DNA absorbent, being competitive with the commercially available magnetic purification kits.


Subject(s)
Nanocomposites , Bridged Bicyclo Compounds, Heterocyclic , DNA/genetics , Polymers , Spectroscopy, Fourier Transform Infrared
4.
Talanta ; 234: 122636, 2021 Nov 01.
Article in English | MEDLINE | ID: mdl-34364445

ABSTRACT

We report the preparation of flexible polystyrene/polypyrrole (PS/PPy) mats and their successful use as a resistive humidity sensor. These composite membranes were prepared by first obtaining PS films through the electrospinning technique, and then incorporating PPy chains by an in situ chemical polymerization of the pyrrole monomer. The PS fibers were homogeneously distributed, with diameters that obeyed a normal distribution with an average value of (1.04 ± 0.12) µm. The deposition of conducting PPy chains on the surface of the PS fibers was confirmed after characterizing the PS/PPy mats by Fourier transform infrared (FTIR) spectroscopy, scanning electron microscopy (SEM), contact angle measurements, and electrochemical impedance spectroscopy (EIS). When used as humidity sensors, the PS/PPy mats exhibited a sensor response of 128.6%, with fast response ((54.9 ± 3.5)s) and recovery times ((76.8 ± 11.1)s), and stable response under different humidity conditions over several days. These performance characteristics compare favorably to those of previous resistive humidity sensors discussed in the literature.


Subject(s)
Polymers , Pyrroles , Humidity , Polystyrenes
5.
J Environ Sci (China) ; 100: 62-73, 2021 Feb.
Article in English | MEDLINE | ID: mdl-33279054

ABSTRACT

We report the preparation of poly(3,4-ethylene dioxythiophene) (PEDOT)-modified polyvinylidene fluoride electrospun fibers and their use as a novel adsorbent material for the removal of the anionic dye Methyl Orange (MO) from aqueous media. This novel adsorbent material can be used to selectively remove MO on a wide pH range (3.0-10.0), with a maximum capacity of 143.8 mg/g at pH 3.0. When used in a recirculating filtration system, the maximum absorption capacity was reached in a shorter time (20 min) than that observed for batch mode experiments (360 min). Based on the analyses of the kinetics and adsorption isotherm data, one can conclude that the predominant mechanism of interaction between the membrane and the dissolved dye molecules is electrostatic. Besides, considering the estimated values for the Gibbs energy, and entropy and enthalpy changes, it was established that the adsorption process is spontaneous and occurs in an endothermic manner. The good mechanical and environmental stability of these membranes allowed their use in at least 20 consecutive adsorption/desorption cycles, without significant loss of their characteristics. We suggest that the physical-chemical characteristics of PEDOT make these hybrid mats a promising adsorbent material for use in water remediation protocols and effluent treatment systems.


Subject(s)
Water Pollutants, Chemical , Adsorption , Azo Compounds , Bridged Bicyclo Compounds, Heterocyclic , Hydrogen-Ion Concentration , Kinetics , Polymers , Polyvinyls , Thermodynamics
6.
Langmuir ; 36(11): 2920-2929, 2020 03 24.
Article in English | MEDLINE | ID: mdl-32119558

ABSTRACT

Phosphate ions perform a variety of functions in metabolic processes and are essential for all living organisms. The determination of the concentration of phosphate ions is useful in clinical diagnosis of various diseases as an inadequate phosphate level could lead to many health problems. In the search for a cost-effective method of fast monitoring, we investigated the use of cobalt ferrite nanoparticles (CoFeNPs) in the selective recognition of phosphate ions dissolved in aqueous media and more complex samples, such as human blood serum. We prepared these NPs by a chemical coprecipitation route and subjected them to annealing at 600 °C for 1 h. The successful formation of the NPs was confirmed by Fourier transform infrared spectroscopy, X-ray diffraction, transmission electron microscopy, and hysteresis loop measurements. The NPs exhibited a ferrimagnetic behavior, a spinel-type crystalline structure, and hexagonal shape in the nanoscale range. We demonstrated that CoFeNPs containing immobilized fluorescent-labeled single-chain DNA (ssDNA*) probes can be applied for the fast selective detection of phosphate ions dissolved in a liquid medium. We have explored the fact that phosphate groups can displace ssDNA* probes attached to the nanoparticles, therefore causing a perceptible change in the fluorescence signal of the supernatant liquid. This detection method has been tested for the sensing of phosphate ions present both in aqueous solutions and in biological samples, with excellent selectivity and a low limit of detection (∼1.75 nM).

7.
Anal Chim Acta ; 1047: 214-224, 2019 Jan 24.
Article in English | MEDLINE | ID: mdl-30567653

ABSTRACT

When fluorophores attach to nanostructured films of intrinsically conducting polymers (ICPs), a quenching of their fluorescence may occur. We have exploited these characteristics for the development of polymeric films that can be used in a simple and efficient molecular diagnosis protocol based on the selective detection of nucleic acids. Our procedure rests on the fact that the fluorescence of 6-carboxyfluorescein-labeled single-stranded DNA (FAM-ssDNA) probes is quenched upon their immobilization on nanostructured ICP - polypyrrole (PPY) and polyaniline (PANI) - films deposited on polyethylene terephthalate (PET) substrates. Hybridization occurs whenever a sample with the complementary sequence is brought in contact with the immobilized probe, with the newly formed ds-DNA chains detaching from the flexible polymeric film and causing the restoration of the fluorescence. This sensing system exhibits a low background signal that depends on both the thickness and hydrophobicity of the films. As a model system, we used a FAM-ssDNA probe specific for the Leishmania infantum parasite. The results confirm this procedure as a simple, fast and highly sensitive scheme for the recognition of the target DNA, with a detection limit of the 1.1 nM and 1.3 nM for the PPY/PET and PANI/PET films, respectively. In addition, this biosensor has excellent stability and exhibits a good and reproducible performance even when used for the direct detection of ssDNA in relatively complex biological samples.


Subject(s)
Aniline Compounds/chemistry , Biosensing Techniques/methods , DNA Probes/chemistry , DNA/analysis , DNA/chemistry , Polymers/chemistry , Pyrroles/chemistry , DNA/genetics , DNA Probes/genetics , Electric Conductivity , Fluoresceins/chemistry , Fluorescence , Fluorescent Dyes/chemistry , Leishmania infantum/genetics , Limit of Detection , Nucleic Acid Hybridization , Reproducibility of Results , Spectrometry, Fluorescence/methods
8.
J Chem Phys ; 148(19): 194304, 2018 May 21.
Article in English | MEDLINE | ID: mdl-30307241

ABSTRACT

We examine the relative contribution of ballistic and elastic cotunneling mechanisms to the charge transport through a single decanedithiol molecule linked to two terminal clusters of gold atoms. For this, we first introduced a conceptual model that permits a generalization of the Breit-Wigner scattering formalism where the cation, anion, and neutral forms of the molecule can participate with different probabilities of the charge transfer process, but in a simultaneous manner. We used a density functional theory treatment and considered the fixed geometry of each charge state to calculate the corresponding eigenvalues and eigenvectors of the extended system for different values of the external electric field. We have found that for the ballistic transport the HOMO and LUMO of the neutral species play a key role, while the charged states give a negligible contribution. On the other hand, an elastic cotunneling charge transfer can occur whenever a molecular orbital (MO) of the cation or anion species, even if localized in just one side of the molecule-gold clusters complex, has energy close to that of a delocalized MO of the neutral species. Under these conditions, a conduction channel is formed throughout the entire system, in a process that is controlled by the degree of resonance between the MOs involved. Our results indicate that while different charge transfer mechanisms contribute to the overall charge transport, quantum effects such as avoided-crossing situations between relevant frontier MOs can be of special importance. In these specific situations, the interchange of spatial localization of two MOs involved in the crossing can open a new channel of charge transfer that otherwise would not be available.

9.
Colloids Surf B Biointerfaces ; 160: 724-731, 2017 Dec 01.
Article in English | MEDLINE | ID: mdl-29035820

ABSTRACT

Electrical impedance spectroscopy (EIS) appears a promising label-free methodology for the investigation of processes related to the aggregation of macromolecules in solution. Here, we explore the EIS technique as a convenient tool for studying the irreversible aggregation of human insulin and describing its corresponding fibrillation kinetics. The in situ measurement of the electrical response of pure insulin solutions at 60°C allows for the real-time monitoring of the protein fibrillation as a function of the incubation time. The fitting of the EIS data through an equivalent circuit based on a constant phase element provides a simple set of electric parameters whose abrupt changes can be associated to transitions occurring in the organization of the macromolecules. For establishing the reliability of the method proposed, we have compared the protein aggregation profile collected from the EIS data to that obtained from a conventional fluorescence methodology where Thioflavin T (ThT) is used as a dye probe. The description of the fibrillation process is quite similar in both cases, since characteristic times of the same order were found for the consecutive processes associated to the initial lag phase of insulin fibrillation, to the rapid growth of amyloidal aggregates and to the final saturation step. Our results suggest that in situ EIS can be considered as a promising approach for the real-time label-free monitoring of protein fibril formation.


Subject(s)
Amyloid/chemistry , Dielectric Spectroscopy/methods , Insulin/chemistry , Protein Aggregation, Pathological , Humans , Kinetics , Protein Aggregates , Reproducibility of Results , Time Factors
10.
Colloids Surf B Biointerfaces ; 148: 576-584, 2016 Dec 01.
Article in English | MEDLINE | ID: mdl-27693719

ABSTRACT

In the last ten years, conjugated polymers started to be used in the immobilization of nucleic acids via non-covalent interactions. In the present study, we describe the construction and use of an electrochemical DNA biosensor based on a nanostructured polyaniline-gold composite, specifically developed for the detection of the BCR/ABL chimeric oncogene. This chromosome translocation is used as a biomarker to confirm the clinical diagnosis of both chronic myelogenous leukemia (CML) and acute lymphocytic leukemia (ALL). The working principle of the biosensor rests on measuring the conductivity resulting from the non-covalent interactions between the hybrid nanocomposite and the DNA probe. The nanostructured platform exhibits a large surface area that enhances the conductivity. Positive cases, which result from the hybridization between DNA probe and targeted gene, induce changes in the amperometric current and in the charge transfer resistance (RCT) responses. Atomic force microscopy (AFM) images showed changes in the genosensor surface after exposure to cDNA sample of patient with leukemia, evidencing the hybridization process. This new hybrid sensing-platform displayed high specificity and selectivity, and its detection limit is estimated to be as low as 69.4 aM. The biosensor showed excellent analytical performance for the detection of the BCR/ABL oncogene in clinical samples of patients with leukemia. Hence, this electrochemical sensor appears as a simple and attractive tool for the molecular diagnosis of the BCR/ABL oncogene even in early-stage cases of leukemia and for the monitoring of minimum levels of residual disease.


Subject(s)
Fusion Proteins, bcr-abl/genetics , Metal Nanoparticles/chemistry , Nanocomposites/chemistry , Polymers/chemistry , Aniline Compounds/chemistry , Biosensing Techniques/methods , DNA Probes/chemistry , DNA Probes/genetics , DNA, Neoplasm/chemistry , DNA, Neoplasm/genetics , Electric Conductivity , Electrochemical Techniques/methods , Gold/chemistry , Humans , Leukemia/diagnosis , Leukemia/genetics , Microscopy, Atomic Force , Polymerase Chain Reaction , Reproducibility of Results , Sensitivity and Specificity
11.
Colloids Surf B Biointerfaces ; 135: 833-839, 2015 Nov 01.
Article in English | MEDLINE | ID: mdl-25847459

ABSTRACT

Unusual methods for specific detection of pathogenic bacteria are becoming key points for control and identification of problems related to health and (bio)safety. In this context, this work aims to propose a new approach for the development of nanostructured biosensors based on carbon nanotubes (CNTs) and antimicrobial peptides for bacterial detection. Firstly, the antimicrobial peptide clavanin A (ClavA) was chemically immobilized on CNTs and surface-immobilized ClavA was used to detect Klebsiella pneumoniae, Enterococcus faecalis, Escherichia coli and Bacillus subtilis in a direct assay format. We used electrochemical impedance spectroscopy technique to evaluate the effectiveness and sensitivity of the ClavA-based biosensors by measuring the modifications in their electrochemical responses before and after incubation in presence of different bacteria concentrations. The biosensor was able to discriminate between bacteria concentrations in the 10(2)-10(6)CFU mL(-1) range. Atomic force microscopy analysis confirmed the biosensor functionality for bacterial recognition. This new sensor system was capable of differentiating between Gram-positive and Gram-negative bacteria, since ClavA showed different affinities toward the pathogenic bacteria species.


Subject(s)
Bacteria/isolation & purification , Blood Proteins/chemistry , Nanostructures , Nanotubes, Carbon , Bacteria/classification , Biosensing Techniques , Dielectric Spectroscopy , Microscopy, Atomic Force
12.
ACS Appl Mater Interfaces ; 7(13): 7231-40, 2015 Apr 08.
Article in English | MEDLINE | ID: mdl-25761543

ABSTRACT

We describe the in situ preparation of a multipurpose hierarchical polyaniline-polystyrene (PANI-PS) composite based in the chemical polymerization of PANI on nonwoven (NW) electrospun PS mats. We performed a detailed study of the properties of these materials to select the best strategies to incorporate PANI chains into pristine NW PS mats without compromising the original porosity and mechanical flexibility of the matrices. The resulting composites presented nanostructured PANI chains highly dispersed in the interior of the NW PS mat and showed good electrical properties and surface-wetting characteristics that could be easily controlled. In particular, we show that these NW PANI-PS mats exhibit interesting properties in their interaction with heavy metal ions. For instance, their high adsorption capacities toward dispersed Hg(II), Cd(II), Pb(II), Cr(VI), and Cu(II) ions make them promising materials for water remediation, by providing a simple manner of collecting and removing these metals from aqueous systems. In fact, the NW electrospun mats here presented do not suffer from the usual limitations found in materials commonly employed as adsorbents, such as a tendency to agglomerate or accumulate in the environment because of difficulties of properly recovering them after use. To better understand the nature of each pairwise metal-PANI interaction, we performed a thorough investigation of the optical and electrical changes induced by the metal adsorption in the NW PANI-PS mats. As a consequence of their interaction with the metal ions, the visual aspect of the mats change, a fact more evident in the case of Cr(VI) removal, when the matrices vary their color from green to purple. These changes are related to the variation of the oxidation state of the PANI chains: as the ion metals are progressively adsorbed into the mat, they promote the conversion in varying degrees of the PANI chains from salt emeraldine to the pernigraniline form, and the mats become more resistive. We implemented an electrical impedance investigation of the charge transport characteristics of NW PANI-PS mat, and the results indicate that they are sensitive to the type of metal ion adsorbed and that the amount of ions adsorbed in each case is mostly related to the standard electrode potential of the metal considered.


Subject(s)
Aniline Compounds/chemistry , Metals, Heavy/isolation & purification , Nanocomposites/chemistry , Nanofibers/chemistry , Polystyrenes/chemistry , Water Pollutants, Chemical/isolation & purification , Adsorption , Materials Testing , Metals, Heavy/chemistry , Nanocomposites/ultrastructure , Nanofibers/ultrastructure , Particle Size , Porosity , Tensile Strength , Water Pollutants, Chemical/chemistry , Water Purification/methods
13.
J Chem Phys ; 141(12): 124712, 2014 Sep 28.
Article in English | MEDLINE | ID: mdl-25273466

ABSTRACT

Based on a quantum chemical valence formalism that allows the rigorous construction of best-localized molecular orbitals on specific parts of an extended system, we examined the separability of individual components of model systems relevant to the description of electron transport in molecular devices. We started by examining how to construct the maximally localized electronic density at the tip of a realistic model of a gold electrode. By varying the number of gold atoms included in the local region where to project the total electronic density, we quantitatively assess how many molecular orbitals are entirely localized in that region. We then considered a 1,4-benzene-di-thiol molecule connected to two model gold electrodes and examined how to localize the electronic density of the total system in the extended molecule, a fractional entity comprising the organic molecule plus an increasing number of the closest metal atoms. We were able to identify in a rigorous manner the existence of three physically different electronic populations, each one corresponding to a distinct set of molecular orbitals. First, there are those entirely localized in the extended molecule, then there is a second group of those completely distributed in the gold atoms external to that region, and, finally, there are those delocalized over the entire system. This latter group can be associated to the shared electronic population between the extended molecule and the rest of the system. We suggest that the treatment here presented could be useful in the theoretical analysis of the electronic transport in nanodevices whenever the use of localized molecular states are required by the physics of the specific problem, such as in cases of weak coupling and super-exchange limits.

14.
Biochim Biophys Acta ; 1838(11): 2899-909, 2014 Nov.
Article in English | MEDLINE | ID: mdl-25109938

ABSTRACT

This work aims to investigate the possible mechanism of action of the homologue peptide Pa-MAP based on the Antarctic fish Pleuronectes americanus, through a study by electrical impedance spectroscopy (EIS) of models of bilayer lipid membranes supported (BLM-s) on solid substrates. For comparison and validation of the data obtained by EIS, we also conducted a study evaluating the human peptide LL-37, whose mechanism of action is well described in the literature: its dielectric response was found to be similar to that of Pa-MAP. The results obtained indicate that Pa-MAP has a good potential for use as a membrane-disrupting peptide and also suggest that the corresponding mechanism of action occurs according to the carpet model followed by a detergent-like effect. The addition of either one of these peptides at different concentrations resulted in a drastic decrease in the membrane's resistance, after just 1min of exposure. Additionally, it was seen that the peptides Pa-MAP and LL-37 may act on membranes with different charges, in an indication of a possible broad spectrum antimicrobial activity. These interactions with different membrane compositions have been attributed to the peptides' structure, mainly due to the presence of many hydrophobic amino acid residues, as observed by in silico studies. Here, we describe the Pa-MAP mechanism of action for the first time. Furthermore, we report the data demonstrating that EIS can be used for studies of peptide-membranes interaction, even when small changes on the surface of the electrode can be detected.

15.
Colloids Surf B Biointerfaces ; 117: 549-54, 2014 May 01.
Article in English | MEDLINE | ID: mdl-24447687

ABSTRACT

In this paper, we report the use of Concanavalin A (ConA) and electrosynthesized polyaniline (PANI) thin films for the development of a new electrochemical sensor that allows the specific detection of two bacterial toxins: lipopolysaccharide (LPS) from Escherichia coli and lipoteichoic acid from Staphylococcus aureus. The impedimetric sensor is fabricated by using glutaraldehyde to self-assemble ConA lectin on PANI-modified steel electrodes through covalent binding. ConA acts as a recognition element for bacterial toxins. Electrical impedance spectroscopy (EIS) and scanning electron microscope (SEM) were applied to characterize the assembly process on the modified electrode. The EIS measurements revealed that the resistance charge transfer (RCT) of the electrode/electrolyte interface increases considerably after the ConA lectin interacts with specific carbohydrate moieties present in the molecule of the bacterial toxin. Our results showed that the ConA lectin retained its activity after immobilization on the PANI surface and also the existence of electrochemical impedance response of the bioelectrode which is linear to the extent of the lectin-toxin interaction, with maximum values of RCT for E. coli (14.40 kΩ), and S. aureus (17.80 kΩ). We have observed that electrosynthesized PANI is an excellent support layer for the covalent binding of lectins on the electrode surface. Thus, the recognition system provides an appropriate biomimetic interface for detection of specific constituents of gram-positive and gram-negative bacteria.


Subject(s)
Aniline Compounds/chemistry , Bacterial Toxins/analysis , Biosensing Techniques/methods , Concanavalin A/chemistry , Dielectric Spectroscopy/methods , Escherichia coli/chemistry , Microscopy, Electron, Scanning , Staphylococcus aureus/chemistry
16.
Curr Protein Pept Sci ; 14(7): 543-55, 2013 Nov.
Article in English | MEDLINE | ID: mdl-23968347

ABSTRACT

Antimicrobial peptides (AMPs) have been isolated from a wide variety of organisms that include microorganisms, plants, insects, frogs and mammals. As part of the innate immune system expressed in many tissues, AMPs are able to provide protection against invasion of foreign microorganisms and exhibit a broad spectrum of activity against bacteria, fungi and/or virus. Non-AMPs cell-penetrating peptides have been used as carriers for overcoming the membrane barrier and helping in the delivery of various molecules into the cell. Physicochemical peptide-lipid interactions studies can provide us with reliable molecular information about microbe defense response, including the elucidation of the prevailing mechanisms of its action, such as the barrel-stave, toroidal pore, carpet and detergent-like models. In this paper, we present an overview of the peptide-lipid mechanisms of interaction as well as discuss alternative techniques that could help to elucidate the peptides functionality. Quartz crystal microbalance (QCM), surface plasmon resonance (SPR) spectroscopy and electrochemical impedance spectroscopy (EIS) are useful techniques to investigate in details of the peptide-membrane interaction. The techniques here discussed could also offer specific and low-cost methods that can to shed some light over the different modes of action of AMPs, contributing to the development of drugs against infectious diseases.


Subject(s)
Biophysics/methods , Cell Membrane/metabolism , Optical Phenomena , Peptides/metabolism , Amino Acid Sequence , Electric Impedance , Molecular Docking Simulation , Molecular Sequence Data , Peptides/chemistry
17.
J Colloid Interface Sci ; 396: 258-63, 2013 Apr 15.
Article in English | MEDLINE | ID: mdl-23465186

ABSTRACT

We describe a new DNA biosensor for the detection of toxigenic Penicillium sclerotigenum in pure culture or infected yams. The P. sclerotigenum detection takes place on a self-assembled monolayer of a (magnetite)/(poly(allylamine hydrochloride)) (Fe3O4-PAH) composite that serves as an anchoring layer for the DNA hybridization interaction. Electrical impedance spectroscopy (EIS) was used to evaluate and quantify the hybridization degree. The Fe3O4-PAH composite is a good platform for the immobilization of biomolecules, due to the presence of many possible binding sites for nucleotides and to its large surface-to-volume ratio and good biocompatibility. The biosensor was capable of not only qualitatively detecting the presence of the fungus genome at low concentrations, but also shown a good quantitative impedimetric response its electrical resistance was monitored along the time of exposure. A Fe3O4-PAH-probe biosensor would require only small volumes and low concentrations of the analyte when used, for instance, in detecting P. sclerotigenum contamination of food, besides presenting many comparative advantages, such as selectivity, specificity and reproducibility, relative to alternative techniques.


Subject(s)
Biosensing Techniques/methods , Dioscorea/microbiology , Penicillium/isolation & purification , Dielectric Spectroscopy , Ferrosoferric Oxide , Metal Nanoparticles , Nucleic Acid Hybridization , Polyamines/chemistry , Reproducibility of Results , Sensitivity and Specificity
18.
J Colloid Interface Sci ; 362(2): 517-23, 2011 Oct 15.
Article in English | MEDLINE | ID: mdl-21816406

ABSTRACT

A sensitive and selective biosensor for dengue serotyping was successfully developed. The biosensor uses a novel gold nanoparticles-polyaniline hybrid composite (AuNpPANI) for the immobilization of Bauhinia monandra lectin (BmoLL). The nanocomposite was applied to a bare gold electrode surface by chemical adsorption, and BmoLL was subsequently electrostatically adsorbed to the nanocomposite-modified surface. Atomic force microscopy (AFM), cyclic voltammetry (CV) and electrochemical impedance (EI) techniques were applied to evaluate the immobilization of BmoLL on AuNpPANI. The AFM images for AuNpPANI-BmoLL-DEN systems indicate a homogenous, compact and dense film of the conjugate. In the EI analyses, an obvious difference of the electron transfer resistance between the AuNpPANI-modified electrode and the bare gold electrode was observed. Among three dengue serotypes studied, dengue serotype 2 (DEN2) has higher values for R(CT), and lower values for both n and Q. These are indications of a larger blocking effect and smaller capacitive dispersion, resulting from the higher agglutination of glycoproteins from the DEN2 sera. The selective BmoLL recognition for various dengue serotypes may be attributed to different patterns of glycoproteins in the sera produced by the glycoprotein immunoresponse from patients infected by the dengue virus.


Subject(s)
Dengue Virus/classification , Dengue/diagnosis , Electrochemical Techniques/instrumentation , Lectins , Nanocomposites , Bauhinia/chemistry , Biosensing Techniques/methods , Electrodes , Gold , Humans , Nanocomposites/chemistry , Serotyping
19.
Colloids Surf B Biointerfaces ; 88(1): 100-7, 2011 Nov 01.
Article in English | MEDLINE | ID: mdl-21752609

ABSTRACT

This work is dedicated to the investigation of the prevailing molecular interactions between Cratylia mollis (Cramoll) and Concanavalin A (Con A) lectins and ionic (sodium dodecylsulfate, SDS) and non-ionic (Triton X-100, TX-100) surfactants, where we have used electrical impedance spectroscopy to map the dielectric characteristics of mixed lectin/surfactant solutions. The disorder induced in the lectin conformation is proportional to the extent of the access of the surfactant to the fluorophore present in the protein, resulting in its progressive unfolding. The complete unfolding of the lectin is associated to the formation of micelles in the core of the protein, each one of them containing a large number of detergent molecules, and therefore the process can be accompanied by measuring the electrical response of the binary surfactant/lectin system. For instance, the change in the real part of the impedance as a function of the relative concentration of the surfactant in the binary solution exhibits a breaking in its linear behavior that can be taken as indicative of a qualitative change in the environment surrounding the protein residue. We consider these results strong evidence in favor of using impedance spectroscopy methods for the analysis of protein-surfactant associations and for the characterization of the interactions that must prevail when the protein unfolds as the relative surfactant concentration is increased in aqueous solutions of these binary systems.


Subject(s)
Concanavalin A/chemistry , Dielectric Spectroscopy/methods , Electric Impedance , Octoxynol/chemistry , Protein Unfolding , Sodium Dodecyl Sulfate/chemistry , Spectrometry, Fluorescence , Surface-Active Agents/chemistry
20.
J Phys Chem A ; 115(27): 7994-8002, 2011 Jul 14.
Article in English | MEDLINE | ID: mdl-21678926

ABSTRACT

The structure and spectroscopic properties of a diluted compound can be deeply affected by its interaction with the neighboring molecules of the solvent, and the associated solvatochromism is an effect that becomes more noticeable with the increase in both the dipole moment of the solute and the polarity of the medium. The correct description of the complex set of interactions that prevail in the solvation process remains a challenge for theoreticians not only when interpreting an observed behavior but also when considering the possible existence of novel properties in untested solute-solvent systems. On the basis of an ab initio study, we examine here how the presence of solvents of different polarities should affect the electronic properties of a family of molecules, formally related to Betaine-30 (aka Reichardt's dye), whose donor (D) and acceptor (A) groups are terminally connected to conjugated chains of different sizes. Because these molecules exhibit elevated ground-state dipole moment that should strongly interact with molecules of a polar solvent, a large hypsochromic shift is predicted for them. However, in a recent gas-phase study of these molecules, we have established the existence of an "inversion" in the spatial localization of their frontier orbitals when the size of the conjugated bridge connecting the D and A groups is progressively increased. This fact has led us to suggest that the increase in size of dissolved betaines should be accompanied by a large variation in their solvatochromic properties. In this work, we first use the self-consistent reaction field approach at the configuration interaction level to estimate the expected bathochromic shift in the absorption spectra (positive solvatochromism) in the largest members of the investigated betaine family when dissolved in different low polarity solvents and then discuss the conformational changes as a consequence of the solute--solvent interactions. We then use these results to interpret the observed solvatochromic properties of push--pull molecules of varying size and discuss the corresponding implications on their photochemical properties.


Subject(s)
Betaine/chemistry , Models, Chemical , Solvents/chemistry , Molecular Conformation , Molecular Weight , Photochemical Processes
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