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1.
Talanta ; 273: 125881, 2024 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-38492283

RESUMO

This work describes fabrication of gold electrodes modified with peptide conjugate DAL-PEG-DK5-PEG-OH that enables ultra-sensitive detection of lipopolysaccharide (LPS) isolated from the reference strain of Escherichia coli O26:B6. The initial step of the established procedure implies immobilization of the fully protected DAL-PEG-DK5-PEG-OH peptide on the surface of the gold electrode previously modified by cysteamine. Then side chain- and Fmoc-deprotection was performed in situ on the electrode surface, followed by its incubation in 1 % of BSA solution to block non-specific bindings sites before LPS detection. The efficiency of the modification was confirmed by X-ray Photoelectron Spectroscopy (XPS) measurements. Additionally, the cyclic voltammetry (CV) and electrochemical impendance spectroscopy (EIS) were employed to monitor the effectiveness of each step of the modification. The obtained results confirmed that the presence of the surface-attached covalently bound peptide DAL-PEG-DK5-PEG-OH enables LPS detection by means of CV technique within the range from 5 × 10-13 to 5 × 10-4 g/mL in PBS solution. The established limit of detection (LOD) for EIS measurements was 4.93 × 10-21 g/mL with wide linear detection range from 5 × 10-21 to 5 × 10-14 g/mL in PBS solution. Furthermore, we confirmed the ability of the electrode to detect LPS in a complex biological samples, like mouse urine and human serum. The effectiveness of the electrodes in identifying LPS in both urine and serum matrices was confirmed for samples containing LPS at both 2.5 × 10-15 g/mL and 2.5 × 10-9 g/mL.


Assuntos
Técnicas Biossensoriais , Lipopolissacarídeos , Animais , Camundongos , Humanos , Ouro/química , Peptídeos Antimicrobianos , Endotoxinas , Eletrodos , Peptídeos , Técnicas Eletroquímicas/métodos , Técnicas Biossensoriais/métodos
2.
Biochim Biophys Acta Biomembr ; 1866(3): 184266, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38151198

RESUMO

This work describes the electrochemical studies on the interactions between V57G mutant of human cystatin C (hCC V57G) and membrane bilayer immobilized on the surface of a gold electrode. The electrode was modified with 6-mercaptohexan-1-ol (MCH) and 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC). DMPC was used as a membrane mimetic for monitoring electrochemical changes resulting from the interactions between the functionalized electrode surface and human cystatin C. The interactions between the modified electrode and hCC V57G were investigated by cyclic voltammetry and electrochemical impedance spectroscopy in a phosphate buffered saline (PBS) containing Fe(CN)63-/4- as a redox probe. The electrochemical measurements confirm that fabricated electrode is sensitive to hCC V57G at the concentration of 1 × 10-14 M. The incubation studies carried out at higher concentrations resulted in insignificant changes observed in cyclic voltammetry and electrochemical impedance spectroscopy measurements. The calculated values of surface coverage θR confirm that the electrode is equally covered at higher concentrations of hCC V57G. Measurements of wettability and surface free energy made it possible to determine the influence of individual structural elements of the modified gold electrode on its properties, and thus allowed to understand the nature of the interactions. Contact angle values confirmed the results obtained during electrochemical measurements, indicating the sensitivity of the electrode towards hCC V57G at the concentration of 1 × 10-14 M. In addition, the XPS spectra confirmed the successful anchoring of hCC V57G to the DMPC-functionalized surface.


Assuntos
Bicamadas Lipídicas , Fosfolipídeos , Humanos , Bicamadas Lipídicas/química , Dimiristoilfosfatidilcolina/química , Ouro/química , Cistatina C , Eletrodos
3.
Chemosphere ; 304: 135381, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-35716709

RESUMO

Carbon felts are flexible and scalable, have high specific areas, and are highly conductive materials that fit the requirements for both anodes and cathodes in advanced electrocatalytic processes. Advanced oxidative modification processes (thermal, chemical, and plasma-chemical) were applied to carbon felt anodes to enhance their efficiency towards electro-oxidation. The modification of the porous anodes results in increased kinetics of acetaminophen degradation in aqueous environments. The utilised oxidation techniques deliver single-step, straightforward, eco-friendly, and stable physiochemical reformation of carbon felt surfaces. The modifications caused minor changes in both the specific surface area and total pore volume corresponding with the surface morphology. A pristine carbon felt electrode was capable of decomposing up to 70% of the acetaminophen in a 240 min electrolysis process, while the oxygen-plasma treated electrode achieved a removal yield of 99.9% estimated utilising HPLC-UV-Vis. Here, the electro-induced incineration kinetics of acetaminophen resulted in a rate constant of 1.54 h-1, with the second-best result of 0.59 h-1 after oxidation in 30% H2O2. The kinetics of acetaminophen removal was synergistically studied by spectroscopic and electrochemical techniques, revealing various reaction pathways attributed to the formation of intermediate compounds such as p-aminophenol and others. The enhancement of the electrochemical oxidation rates towards acetaminophen was attributed to the appearance of surface carbonyl species. Our results indicate that the best-performing plasma-chemical treated CFE follows a heterogeneous mechanism with only approx. 40% removal due to direct electro-oxidation. The degradation mechanism of acetaminophen at the treated carbon felt anodes was proposed based on the detected intermediate products. Estimation of the cost-effectiveness of removal processes, in terms of energy consumption, was also elaborated. Although the study was focussed on acetaminophen, the achieved results could be adapted to also process emerging, hazardous pollutant groups such as anti-inflammatory pharmaceuticals.


Assuntos
Acetaminofen , Poluentes Químicos da Água , Acetaminofen/química , Fibra de Carbono , Carvão Vegetal , Eletrodos , Peróxido de Hidrogênio/química , Oxirredução , Oxigênio , Água , Poluentes Químicos da Água/química
4.
Materials (Basel) ; 16(1)2022 Dec 29.
Artigo em Inglês | MEDLINE | ID: mdl-36614648

RESUMO

The search for new molecular recognition systems has become the goal of modern electrochemistry. Creating a matrix in which properties can be controlled to obtain a desired analytical signal is an essential part of creating such tools. The aim of this work was to modify the surface of electrodes based on transparent conductive oxides with the use of selected alkoxysilanes (3-aminopropyltrimethoxysilane, trimethoxy(propyl)silane, and trimethoxy(octyl)silane). Electrochemical impedance spectroscopy and cyclic voltammetry techniques, as well as contact angle measurements, were used to determine the properties of the obtained layers. Here, we prove that not only was the structure of alkoxysilanes taken into account but also the conditions of the modification process-reaction conditions (time and temperature), double alkoxysilane modification, and mono- and binary component modification. Our results enabled the identification of the parameters that are important to ensure the effectiveness of the modification process. Moreover, we confirmed that the selection of the correct alkoxysilane allows the surface properties of the electrode material to be controlled and, consequently, the charge transfer process at the electrode/solution interface, hence enabling the creation of selective molecular recognition systems.

5.
Materials (Basel) ; 14(24)2021 Dec 14.
Artigo em Inglês | MEDLINE | ID: mdl-34947318

RESUMO

Over the past few decades, nanoparticles of iron oxide Fe3O4 (magnetite) gained significant attention in both basic studies and many practical applications. Their unique properties such as superparamagnetism, low toxicity, synthesis simplicity, high surface area to volume ratio, simple separation methodology by an external magnetic field, and renewability are the reasons for their successful utilisation in environmental remediation, biomedical, and agricultural applications. Moreover, the magnetite surface modification enables the successful binding of various analytes. In this work, we discuss the usage of core-shell nanoparticles and nanocomposites based on Fe3O4 for the modification of the GC electrode surface. Furthermore, this review focuses on the heavy metal ions electrochemical detection using Fe3O4-based nanoparticles-modified electrodes. Moreover, the most frequently used electrochemical methods, such as differential pulse anodic stripping voltammetry and measurement conditions, including deposition potential, deposition time, and electrolyte selection, are discussed.

6.
J Mech Behav Biomed Mater ; 121: 104648, 2021 09.
Artigo em Inglês | MEDLINE | ID: mdl-34153624

RESUMO

Contemporary prosthetic materials are characterized by highly specific preparation for a given application. This means that at the stage of their creation, not only their function is taken into account, but also the long-term behavior of this material during use. In the case of telescopic crowns, an important factor not yet appearing in the research is the aspect of adhesion force and its dependence on the type of biomaterial, but also the properties of human saliva. The use of artificial saliva, which creates a lubricating layer, reduces the wear on the surface of the telescopic crowns by reducing friction. The impact of artificial saliva on the formation of chemical bonds between prosthetic elements, thus contributing to the so-called retention force has not yet been studied. In this work, two types of measurements of gold telescopic crown materials in the aspect of the adhesion process are presented. Obtained results allowed to fully characterize this phenomenon. We modeled the load force between the microcircuit and the surface under study to suit the conditions between the primary and secondary crowns in the patient's mouth.


Assuntos
Coroas , Revestimento de Dentadura , Materiais Dentários , Planejamento de Prótese Dentária , Fricção , Humanos , Microscopia de Força Atômica
7.
Bioelectrochemistry ; 139: 107742, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33517203

RESUMO

This work describes the modification of a gold electrode with the BMS-8 compound that interacts with the Programmed Death-Ligand 1 (PD-L1), an immune checkpoint protein. The results show that we can confirm the presence of the sPD-L1 in the concentration range of 10-18 to 10-8 M using electrochemical impedance spectroscopy (EIS) with a limit of detection (LOD) of 1.87 × 10-14 M for PD-L1 (S/N = 3.3) and at a concentration of 10-14 M via cyclic voltammetry (CV). Additionally, high-resolution X-ray photoelectron spectroscopy (XPS), contact angle, and surface free energy measurements were applied to confirm the functionalization of the electrode. We investigated the selectivity of the electrode for other proteins: Programmed Death-1 (PD-1), cluster of differentiation 160 (CD160), and B- and T-lymphocyte attenuator (BTLA) at concentrations of 10-8 M. Differentiation between PD-L1 and PD-1 was achieved based on the analysis of the capacitance effect frequency dispersion at the surface of the modified Au electrode with BMS-8 after incubation at various concentrations of PD-L1 and PD-1 proteins in the range of 10-18 to 10-8 M. Significant differences were observed in the heterogeneity of PD-L1 and PD-1. The results of the quasi-capacitance studies demonstrate that BMS-8 strongly and specifically interacts with the PD-L1 protein.


Assuntos
Antígeno B7-H1/análise , Técnicas Biossensoriais/métodos , Espectroscopia Dielétrica/métodos , Técnicas Eletroquímicas/métodos , Neoplasias/diagnóstico , Receptor de Morte Celular Programada 1/análise , Biomarcadores Tumorais/análise , Capacitância Elétrica , Eletrodos , Ouro/química , Humanos , Limite de Detecção , Nanopartículas Metálicas/química , Sensibilidade e Especificidade
8.
Materials (Basel) ; 13(22)2020 Nov 16.
Artigo em Inglês | MEDLINE | ID: mdl-33207805

RESUMO

The present work describes synthesis, characterization, and use of a new dansyl-labelled Ag@SiO2 nanocomposite as an element of a new plasmonic platform to enhance the fluorescence intensity. Keeping in mind that typical surface plasmon resonance (SPR) characteristics of silver nanoparticles coincide well enough with the absorption of dansyl molecules, we used them to build the core of the nanocomposite. Moreover, we utilized 10 nm amino-functionalized silica shell as a separator between silver nanoparticles and the dansyl dye to prevent the dye-to-metal energy transfer. The dansyl group was incorporated into Ag@SiO2 core-shell nanostructures by the reaction of aminopropyltrimethoxysilane with dansyl chloride and we characterized the new dansyl-labelled Ag@SiO2 nanocomposite using transmission electron microscopy (TEM) and Fourier-transform infrared spectroscopy (FTIR). Additionally, water wettability measurements (WWM) were carried out to assess the hydrophobicity and hydrophilicity of the studied surface. We found that the nanocomposite deposited on a semitransparent silver mirror strongly increased the fluorescence intensity of dansyl dye (about 87-fold) compared with the control sample on the glass, proving that the system is a perfect candidate for a sensitive plasmonic platform.

9.
Molecules ; 25(17)2020 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-32882948

RESUMO

In this paper, we described the synthesis procedure of TiO2@SiO2 core-shell modified with 3-(aminopropyl)trimethoxysilane (APTMS). The chemical attachment of Fmoc-glycine (Fmoc-Gly-OH) at the surface of the core-shell structure was performed to determine the amount of active amino groups on the basis of the amount of Fmoc group calculation. We characterized nanostructures using various methods: transmission electron microscope (TEM), scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA) and X-ray photoelectron spectroscopy (XPS) to confirm the modification effectiveness. The ultraviolet-visible spectroscopy (UV-vis) measurement was adopted for the quantitative determination of amino groups present on the TiO2@SiO2 core-shell surface by determination of Fmoc substitution. The nanomaterials were functionalized by Fmoc-Gly-OH and then the fluorenylmethyloxycarbonyl (Fmoc) group was cleaved using 20% (v/v) solution of piperidine in DMF. This reaction led to the formation of a dibenzofulvene-piperidine adduct enabling the estimation of free Fmoc groups by measurement the maximum absorption at 289 and 301 nm using UV-vis spectroscopy. The calculations of Fmoc loading on core-shell materials was performed using different molar absorption coefficient: 5800 and 6089 dm3 × mol-1 × cm-1 for λ = 289 nm and both 7800 and 8021 dm3 × mol-1 × cm-1 for λ = 301 nm. The obtained results indicate that amount of Fmoc groups present on TiO2@SiO2-(CH2)3-NH2 was calculated at 6 to 9 µmol/g. Furthermore, all measurements were compared with Fmoc-Gly-OH used as the model sample.


Assuntos
Fluorenos/química , Glicina/análogos & derivados , Fluorenos/síntese química , Glicina/síntese química , Glicina/química , Nanopartículas/química , Nanopartículas/ultraestrutura , Espectroscopia Fotoeletrônica , Espectroscopia de Infravermelho com Transformada de Fourier , Termogravimetria
10.
Chemosphere ; 260: 127605, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32688319

RESUMO

Polychlorinated dibenzo-p-dioxins, dibenzofurans (PCDD/F) and their precursors - pentachlorophenol (PCP) and triclosan (TCS), constitute a group of persistent, highly toxic multimedia pollutants, being easily transported via atmosphere over long distances, thus particularly threatening to the polar areas. The global fate of PCDD/Fs is temperature-dependent, and their transfer and immobilization at the Poles are described by the grasshopper effect and the cold trap phenomenon. The aim of this interdisciplinary study was to perform a preliminary assessment of the present state of pollution of Arctic and Antarctic marine sediments by PCP and TCS along with determination of PCDD/Fs contamination by immunoassay. Sediments from 20 stations were collected during two polar expeditions (2013-2016). The study area covered Hornsund Fjord and the southwest coast of Wedel-Jarlsberg Land (Arctic) - Skodde Bay, Nottingham Bay, Isbjørnhamna Bay and Admiralty Bay (Antarctica) - Suszczewski Cove, Halfmoon Cove and Herve Cove. The studied contaminants were quantified in 60% of the collected sediments, with almost half exceeding the environmentally safe levels according European regulations and worldwide literature. The determined levels of PCP, TCS and PCDD/F in Arctic and Antarctic sediments were to be comparable to those reported in the southern Baltic Sea located in the intense industrialized mid-latitudes. Maximum concentrations were observed in the vicinity of retreating, marine terminating glaciers. This observation confirms reemission of POPs into the global cycle with respect to the worldwide ocean warming. The results of this study should gain attention of the international and regional environmental agencies as well as the main chlorine production decision makers.


Assuntos
Dibenzofuranos/análise , Monitoramento Ambiental , Poluentes Ambientais/análise , Dibenzodioxinas Policloradas/análise , Regiões Antárticas , Benzofuranos/análise , Mudança Climática , Dibenzofuranos Policlorados , Dioxinas , Poluição Ambiental , Sedimentos Geológicos/química , Pentaclorofenol
11.
Opt Express ; 28(11): 15934-15942, 2020 May 25.
Artigo em Inglês | MEDLINE | ID: mdl-32549427

RESUMO

In this work, we present a direct electrochemical biofunctionalization of an indium-tin-oxide-coated lossy-mode resonance optical fiber sensor. The functionalization using a biotin derivative was performed by cyclic voltammetry in a 10 mM biotin hydrazide solution. All stages of the experiment were simultaneously verified with optical and electrochemical techniques. Performed measurements indicate the presence of a poly-biotin layer on the sensor's surface. Furthermore, dual-domain detection of 0.01 and 0.1 mg/mL of avidin confirms the sensor's viability for label-free detection.

12.
Spectrochim Acta A Mol Biomol Spectrosc ; 222: 117226, 2019 Nov 05.
Artigo em Inglês | MEDLINE | ID: mdl-31181504

RESUMO

Six novel amino acid chromophores were synthesized and their spectroscopic, acid-base, and electrochemical properties are discussed in this work. In studied compounds, selected amino acid residues (l-Aspartic acid, l-Glutamic acid, l-Glutamine, l-Histidine, l-Lysine, l-Arginine) are attached to the 1-(piperazine) 9,10-anthraquinone skeleton via the amide bond between the carboxyl group of amino acid and nitrogen atom of the piperazine ring. All derivatives have been characterized using a variety of spectroscopic techniques (mass spectrometry, 1HNMR, UV-Vis, IR spectroscopy), acid-base (electrochemical and UV-Vis) titrations, and cyclic voltammetry methods. Basing on observed experimental effects, supported by quantum chemical simulations, the structure-properties links were established. They are indicative of the specific interactions within and/or in-between amino acid side groups, which are prone to form both, intra- and intermolecular hydrogen bonds as well as electrostatic interactions with the anthraquinone system.


Assuntos
Aminoácidos/química , Antraquinonas/química , Corantes/química , Aminoácidos/síntese química , Antraquinonas/síntese química , Corantes/síntese química , Técnicas Eletroquímicas , Ligação de Hidrogênio , Modelos Moleculares , Prótons , Espectrofotometria Ultravioleta
13.
Sensors (Basel) ; 18(5)2018 Apr 27.
Artigo em Inglês | MEDLINE | ID: mdl-29702595

RESUMO

In this work an application of optical fiber sensors for real-time optical monitoring of electrochemical deposition of ketoprofen during its anodic oxidation is discussed. The sensors were fabricated by reactive magnetron sputtering of indium tin oxide (ITO) on a 2.5 cm-long core of polymer-clad silica fibers. ITO tuned in optical properties and thickness allows for achieving a lossy-mode resonance (LMR) phenomenon and it can be simultaneously applied as an electrode in an electrochemical setup. The ITO-LMR electrode allows for optical monitoring of changes occurring at the electrode during electrochemical processing. The studies have shown that the ITO-LMR sensor’s spectral response strongly depends on electrochemical modification of its surface by ketoprofen. The effect can be applied for real-time detection of ketoprofen. The obtained sensitivities reached over 1400 nm/M (nm·mg−1·L) and 16,400 a.u./M (a.u.·mg−1·L) for resonance wavelength and transmission shifts, respectively. The proposed method is a valuable alternative for the analysis of ketoprofen within the concentration range of 0.25⁻250 μg mL−1, and allows for its determination at therapeutic and toxic levels. The proposed novel sensing approach provides a promising strategy for both optical and electrochemical detection of electrochemical modifications of ITO or its surface by various compounds.


Assuntos
Fibras Ópticas , Técnicas Biossensoriais , Técnicas Eletroquímicas , Cetoprofeno , Compostos de Estanho
14.
Acta Biochim Pol ; 64(4): 693-698, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29247505

RESUMO

The aim of this study was to investigate the soil microbial communities of a phosphogypsum waste heap. The soil microbial community structures can differ over time, as they are affected by the changing environmental conditions caused by a long-term exposure to different kinds of pollutions, like is the case of soil in the post-production waste area in Wislinka (in the northern part of Poland) currently undergoing restoration. Our analyses indicated that the most abundant phyla were Proteobacteria, Acidobacteria, and Actinobacteria, and generally such an abundance is common for most of the studied soils. The most dominant class were Alphaproteobacteria, with their participation in 33.46% of the total reads. Among this class, the most numbered order was Sphingomonadales, whereas among this order the Sphingomonadaceae family was the most abundant one. The Sphingomonadaceae family is currently in the center of interest of many researchers, due to the ability of some of its members to utilize a wide range of naturally occurring organic compounds and many types of environmental contaminants. This kind of knowledge about microbial populations can support efforts in bioremediation and can improve monitoring changes in the contaminated environments.


Assuntos
Sulfato de Cálcio , Microbiota/fisiologia , Fósforo , Microbiologia do Solo , Poluentes do Solo , Sequenciamento de Nucleotídeos em Larga Escala , Microbiota/genética , Polônia , RNA Ribossômico 16S , Solo/química
15.
Sci Rep ; 7(1): 15707, 2017 Nov 16.
Artigo em Inglês | MEDLINE | ID: mdl-29146948

RESUMO

According to the World Health Organization (WHO), almost 2 billion people each year are infected worldwide with flu-like pathogens including influenza. This is a contagious disease caused by viruses belonging to the family Orthomyxoviridae. Employee absenteeism caused by flu infection costs hundreds of millions of dollars every year. To successfully treat influenza virus infections, detection of the virus during the initial development phase of the infection is critical, when tens to hundreds of virus-associated molecules are present in the patient's pharynx. In this study, we describe a novel universal diamond biosensor, which enables the specific detection of the virus at ultralow concentrations, even before any clinical symptoms arise. A diamond electrode is surface-functionalized with polyclonal anti-M1 antibodies, which then serve to identify the universal biomarker for the influenza virus, M1 protein. The absorption of the M1 protein onto anti-M1 sites of the electrode change its electrochemical impedance spectra. We achieved a limit of detection of 1 fg/ml in saliva buffer for the M1 biomarker, which corresponds to 5-10 viruses per sample in 5 minutes. Furthermore, the universality of the assay was confirmed by analyzing different strains of influenza A virus.


Assuntos
Anticorpos/metabolismo , Técnicas Biossensoriais/métodos , Boro/química , Diamante/química , Vírus da Influenza A/isolamento & purificação , Biomarcadores/análise , Espectroscopia Dielétrica , Eletrodos , Humanos , Propriedades de Superfície
16.
World J Microbiol Biotechnol ; 32(10): 163, 2016 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-27562597

RESUMO

Yersinia sp. bacteria owe their viability and pathogenic virulence to the YopH factor, which is a highly active bacterial protein tyrosine phosphatase. Inhibition of YopH phosphatase results in the lack of Yersinia sp. pathogenicity. We have previously described that aurintricarboxylic acid inhibits the activity of YopH at nanomolar concentrations and represents a unique mechanism of YopH inactivation due to a redox process. This work is a continuation of our previous studies. Here we show that modifications of the structure of aurintricarboxylic acid reduce the ability to inactivate YopH and lead to higher cytotoxicity. In the present paper we examine the inhibitory properties of aurintricarboxylic acid analogues, such as eriochrome cyanine R (ECR) and pararosaniline. Computational docking studies we report here indicate that ATA analogues are not precluded to bind in the YopH active site and in all obtained binding conformations ECR and pararosaniline bind to YopH active site. The free binding energy calculations show that ECR has a stronger binding affinity to YopH than pararosaniline, which was confirmed by experimental YopH enzymatic activity studies. We found that ATA analogues can reversibly reduce the enzymatic activity of YopH, but possess weaker inhibitory properties than ATA. The ATA analogues induced inactivation of YopH is probably due to oxidative mechanism, as pretreatment with catalase prevents from inhibition. We also found that ATA analogues significantly decrease the viability of macrophage cells, especially pararosaniline, while ATA reveals only slight effect on cell viability.


Assuntos
Ácido Aurintricarboxílico/análogos & derivados , Proteínas da Membrana Bacteriana Externa/química , Benzenossulfonatos/química , Proteínas Tirosina Fosfatases/química , Corantes de Rosanilina/química , Toluidinas/química , Yersinia/efeitos dos fármacos , Animais , Ácido Aurintricarboxílico/química , Ácido Aurintricarboxílico/farmacologia , Proteínas da Membrana Bacteriana Externa/antagonistas & inibidores , Benzenossulfonatos/farmacologia , Domínio Catalítico/efeitos dos fármacos , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Camundongos , Modelos Moleculares , Simulação de Acoplamento Molecular , Simulação de Dinâmica Molecular , Estrutura Molecular , Ligação Proteica , Proteínas Tirosina Fosfatases/antagonistas & inibidores , Corantes de Rosanilina/farmacologia , Toluidinas/farmacologia , Yersinia/enzimologia
17.
Oncotarget ; 6(21): 18364-73, 2015 Jul 30.
Artigo em Inglês | MEDLINE | ID: mdl-26286963

RESUMO

YopH is a bacterial protein tyrosine phosphatase, which is essential for the viability and pathogenic virulence of the plague-causing Yersinia sp. bacteria. Inactivation of YopH activity would lead to the loss of bacterial pathogenicity. We have studied the inhibitory properties of aurintricarboxylic acid (ATA) against YopH phosphatase and found that at nanomolar concentrations ATA reversibly decreases the activity of YopH. Computational docking studies indicated that in all binding poses ATA binds in the YopH active site. Molecular dynamics simulations showed that in the predicted binding pose, ATA binds to the essential Cys403 and Arg409 residues in the active site and has a stronger binding affinity than the natural substrate (pTyr). The cyclic voltammetry experiments suggest that ATA reacts remarkably strongly with molecular oxygen. Additionally, the electrochemical reduction of ATA in the presence of a negative potential from -2.0 to 2.5 V generates a current signal, which is observed for hydrogen peroxide. Here we showed that ATA indicates a unique mechanism of YopH inactivation due to a redox process. We proposed that the potent inhibitory properties of ATA are a result of its strong binding in the YopH active site and in situ generation of hydrogen peroxide near catalytic cysteine residue.


Assuntos
Ácido Aurintricarboxílico/química , Proteínas da Membrana Bacteriana Externa/química , Proteínas Tirosina Fosfatases/química , Fatores de Virulência/química , Algoritmos , Ácido Aurintricarboxílico/metabolismo , Ácido Aurintricarboxílico/farmacologia , Proteínas da Membrana Bacteriana Externa/antagonistas & inibidores , Proteínas da Membrana Bacteriana Externa/metabolismo , Humanos , Cinética , Conformação Molecular , Simulação de Dinâmica Molecular , Estrutura Molecular , Oxirredução , Peste/microbiologia , Ligação Proteica , Estrutura Terciária de Proteína , Proteínas Tirosina Fosfatases/antagonistas & inibidores , Proteínas Tirosina Fosfatases/metabolismo , Virulência , Fatores de Virulência/antagonistas & inibidores , Fatores de Virulência/metabolismo , Yersinia pestis/metabolismo , Yersinia pestis/patogenicidade , Yersinia pestis/fisiologia
18.
Biometals ; 28(2): 307-20, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25656562

RESUMO

The aim of this work was to find a relationship between physicochemical properties of the oxovanadium(IV) complexes, namely [VO(ODA)(H2O)2], [VO(ODA)(phen)]·1.5H2O and [VO(ODA)(bipy)]·2H2O (ODA = oxydiacetate) as well as [VO(H2O)5](2+), and their biological activity. A potentiometric titration method has been used to characterize the stability of the complexes in aqueous solutions. Furthermore, the reactivity of the complexes towards superoxide free radicals was assessed by employing the NBT assay as well as a cyclic voltammetry (CV) technique. Additionally, the investigations of the antioxidant properties of the complexes were complemented by studying their reactivity towards organic radicals (the ABTS and DPPH tests). Finally, the biological properties of the complexes were investigated in relation to their cytoprotective activity against the oxidative damage generated exogenously by using hydrogen peroxide in the Hippocampal neuronal cell line HT22 (the MTT and LDH tests). The obtained results showed that all the compounds under study display antioxidant properties but a concentration-depended protective effect against the oxidative damage was found for [VO(ODA)(bipy)]·2H2O only.


Assuntos
Sequestradores de Radicais Livres/farmacologia , Fármacos Neuroprotetores/farmacologia , Vanadatos/química , 2,2'-Dipiridil/química , Acetatos/química , Animais , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Avaliação Pré-Clínica de Medicamentos , Estabilidade de Medicamentos , Sequestradores de Radicais Livres/química , Hipocampo/citologia , Concentração de Íons de Hidrogênio , Camundongos , Fármacos Neuroprotetores/química , Fenantrolinas/química , Soluções , Superóxidos/química
19.
Spectrochim Acta A Mol Biomol Spectrosc ; 137: 979-86, 2015 Feb 25.
Artigo em Inglês | MEDLINE | ID: mdl-25282027

RESUMO

A series of novel polyether derivatives of 9,10-anthraquinone (AQ) was synthesized and characterized by means of UV-Vis spectroscopy, acid-base titration and complexometric titration. The results were compared with 1-NEt2AQ and 1-NHEtAQ--model compounds of alkylaminoanthraquinones. Acetonitrile and methanol were used as solvents for determination of spectroscopic and acid-base properties. Complexometric titrations were carried out exclusively in acetonitrile. Spectral characteristic of these compounds strongly depends on pH. Addition of acid causes the decrease of absorption intensity and in some cases also a shift of the visible range band. The weakest base is the compound (2), and the strongest--compound (1), both in methanol and acetonitrile solution. The introduction of an additional substituent in the position 8 of the anthraquinone compound increases its basicity. The presence of metal ions causes changes in intensity of absorption (decrease for compounds (2) and (3) and increase with bathochromic shift for (1) and (4)). For the determination of the coordination properties aluminum (III) ions were chosen. The highest complex stability constant with Al (III) ions is observed for compound (1), and the weakest for compound (3). The elongation of the polyether chain decreases the stability of the complex formed.


Assuntos
Antraquinonas/química , Poliésteres/química , Acetonitrilas/química , Metanol/química , Espectrofotometria Ultravioleta/métodos
20.
Forensic Sci Int ; 232(1-3): e28-32, 2013 Oct 10.
Artigo em Inglês | MEDLINE | ID: mdl-23938207

RESUMO

This article, written as a result of cooperation between a police forensic laboratory and an academic institution, outlines the possibility of applying single-crystal X-ray diffraction analysis as an effective method of identifying designer drugs in forensic analysis. This technique allows crystalline samples to be determined with full assurance about their identity, even in the case of new substances for which no reference standards yet exist. Here, single-crystal X-ray diffraction measurements of single-crystal specimens obtained from two samples were performed. Solution and refinement of the structures demonstrated that the target compounds were metaphedrone and pentedrone hydrochlorides - synthetic cathinone derivatives used as recreational stimulants. In addition to the identification of the title compounds, this paper gives a first report on their crystal structures. Once the CIF-files containing the crystal structure data of the title compounds have been deposited in the Cambridge Structural Database - the world repository of small molecule crystal structures - it will be possible to identify single crystals of the title compounds quickly on the basis of simple parameters (lattice parameters a, b, c, α, ß, γ and unit cell volume). This description of the relationship between the geometrical parameters of moieties and the analysis of intermolecular interactions occurring in crystals of the title compounds extends knowledge about the synthetic derivatives of cathinone and may play a role in future studies, leading to a better understanding of their characteristic properties.

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