RESUMO
Additive manufacturing is a technique that allows the construction of prototypes and has evolved a lot in the last 20 years, innovating industrial fabrication processes in several areas. In chemistry, additive manufacturing has been used in several functionalities, such as microfluidic analytical devices, energy storage devices, and electrochemical sensors. Theophylline and paracetamol are important pharmaceutical drugs where overdosing can cause adverse effects, such as tachycardia, seizures, and even renal failure. Therefore, this paper aims at the development of miniaturized electrochemical sensors using 3D printing and polylactic acid-based conductive carbon black commercial filament for theophylline and paracetamol detection. Electrochemical characterizations of the proposed sensor were performed to prove the functionality of the device. Morphological characterizations were carried out, in which chemical treatment could change the surface structure, causing the improvement of the analytical signal. Thus, the detection of theophylline at a linear range of 5.00-150 µmol L-1 with a limit of detection of 1.2 µmol L-1 was attained, and the detection of paracetamol at a linear range of 1.00-200 µmol L-1 with a limit of detection of 0.370 µmol L-1 was obtained, demonstrating the proposed sensor effectively detected pharmaceutical drugs.
Assuntos
Acetaminofen , Técnicas Eletroquímicas , Poliésteres , Fuligem , Teofilina , Acetaminofen/análise , Fuligem/química , Técnicas Eletroquímicas/métodos , Teofilina/análise , Poliésteres/química , Limite de Detecção , Impressão Tridimensional , MiniaturizaçãoRESUMO
Here, we present the proof-of-concept of a lateral flow assay (LFA) that is capable of detecting small-molecule targets in a noncompetitive manner by deploying a sandwich-type format based on the aptamer kissing complex (AKC) strategy. A fluorescently labeled hairpin aptamer served as the signaling agent, while a specific RNA hairpin grafted onto the strip served as the capture element. The hairpin aptamer switched from an unfolded to a folded form in the presence of the target, resulting in kissing interactions between the loops of the reporter and the capture agents. This design triggered a target-dependent fluorescent signal at the test line. The AKC-based LFA was developed for the detection of adenosine, achieving a detection limit in the micromolar range. The assay revealed the presence of the same analyte in urine. The method also proved effective with another small molecule (theophylline). We believe that the AKC-based LFA approach could overcome many of the shortcomings associated with conventional signal-off methods and competitive processes.
Assuntos
Adenosina , Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Aptâmeros de Nucleotídeos/química , Adenosina/análise , Adenosina/urina , Técnicas Biossensoriais/métodos , Humanos , Teofilina/análise , Teofilina/urina , Limite de Detecção , Corantes Fluorescentes/químicaRESUMO
In vivo sampling and sensitive detection of environmental pollutants and drugs in human body play a crucial role in understanding human health. In this study, in vivo solid-phase microextraction (SPME) swab was fabricated using a SPME fiber and a medical cotton swab for noninvasive sampling and extraction of environmental pollutants and drugs in human oral cavity, nasal cavity and on skin surface. After sampling, SPME was coupled with nano-electrospray ionization mass spectrometry (nanoESI-MS) for desorption, ionization, and detection of the extracted analytes. As a result, limit of detection (LOD) and limit of quantification (LOQ) of nicotine in oral fluid were found to be 1.0 pg/mL (S/N ≥ 3) and 4.0 pg/mL (S/N ≥ 10), respectively. Linear dynamic signal responses of nicotine exhibited excellent linearity (R2 = 0.9996) in human oral fluid ranging from 0.1 to 50 ng/mL. The coefficient of variation (CV) values of SPME swab for five measurements from sample vials and human body were 5.1-6.7% and 22.7-32.6%, respectively. Rapid analysis of a single sample could be completed within 10 min. Overall, our results demonstrated that SPME swab-MS is a promising noninvasive method for enhanced detection of analytes in human body.
Assuntos
Poluentes Ambientais/análise , Boca/química , Nanotecnologia , Pele/química , Microextração em Fase Sólida , Cafeína/análise , Corpo Humano , Humanos , Imidazóis/análise , Nanotecnologia/instrumentação , Nicotina/análise , Microextração em Fase Sólida/instrumentação , Espectrometria de Massas por Ionização por Electrospray/instrumentação , Teobromina/análise , Teofilina/análiseRESUMO
Lanthanum oxide nanomaterials were decorated with carbon black (CB) and grafted with a poly(acrylic acid) nanogel to obtain a composite material (CB-g-PAA/La2O3) for simultaneous determination of acetaminophen (AMP), naproxen (NPX), and theophylline (TPH). The nanogel was synthesized by in-situ free radical polymerization. The composite was dropped onto a glassy carbon electrode (GCE), and the modified GCE displays robust electrocatalytic activity towards AMP, NPX, and TPH, with voltammetric signals that are enhanced compared to a bare GCE. Features of merit for AMP, NPX, and TPH, respectively, include (a) peak potentials of 0.42, 0.85 and 0.12 V (vs. Ag/AgCl), (b) linear ranges from 0.05-887, 0.05-884, and 0.02-888 µM, and (c) detection limits of 20, 35, and 15 nM. The practical applicability of the CB-g-PAA/La2O3/GCE was illustrated by analyzing serum and urine samples. Graphical abstract Schematic presentation of simultaneous electrochemical sensing of acetaminophen (AMP), naproxen (NPX), and theophylline (TPH) in real sample analysis using poly(acrylic acid) nanogel covalently grafted onto a carbon black/La2O3 composite (CB-g-PAA/La2O3/GCE).
Assuntos
Acetaminofen/análise , Resinas Acrílicas/química , Lantânio/química , Nanogéis/química , Naproxeno/análise , Óxidos/química , Fuligem/química , Teofilina/análise , Acetaminofen/sangue , Acetaminofen/urina , Eletroquímica , Eletrodos , Humanos , Modelos Moleculares , Conformação Molecular , Naproxeno/sangue , Naproxeno/urina , Polimerização , Teofilina/sangue , Teofilina/urinaRESUMO
This study describes a universal fluorometric method for sensitive detection of analytes by using aptamers. It is based on the use of graphene oxide (GO) and cryonase-assisted signal amplification. GO is a strong quencher of FAM-labeled nucleic acid probes, while cryonase digests all types of nucleic acid probes. This makes the platform widely applicable to analytes for which the corresponding aptamers are available. Theophylline and ATP were chosen as model analytes. In the absence of targets, dye-labeled aptamers are in a flexible single strand state and adsorb on the GO. As a result, the probes are non-fluorescent due to the efficient quenching of dyes by GO. Upon the addition of a specific target, the aptamer/target complex desorbed from the GO surface and the probe becomes fluorescent. The released complex will immediately become a substrate for cryonase digestion and subsequently releasing the target to bind to another aptamer to initiate the next round of cleavage. This cyclic reaction will repeat again and again until all the related-probes are consumed and all fluorophores light up, resulting in significant fluorescent signal amplification. The detection limits are 47 nM for theophylline and 22.5 nM for ATP. This is much better than that of known methods. The assay requires only mix-and-measure steps that can be accomplished rapidly. In our perception, the detection scheme holds great promise for the design enzyme-aided amplification mechanisms for use in bioanalytical methods. Graphical abstract A cryonase-assisted signal amplification (CASA) method has been developed by using graphene oxide (GO) conjugated with a fluorophore-labeled aptamer for fluorescence signal generation. It has a large scope because it may be applied to numerous analytes.
Assuntos
Trifosfato de Adenosina/análise , Aptâmeros de Nucleotídeos/química , Técnicas Biossensoriais , Grafite/química , Sondas de Ácido Nucleico/química , Teofilina/análise , Trifosfato de Adenosina/química , Fluorescência , Teofilina/químicaRESUMO
Here, we report a three-dimensional (3D) network molecularly imprinted polymer (MIP) on electrode surface to achieve an efficient and specific detection of theophylline in foodstuffs, using theophylline as the template molecule, mesoporous silica nanospheres (MSNs) as the signal transducer to shuttle electrons, and both phenyltriethoxysilane and pyrrole as functional monomers. The electron microscope images reveal the presence of well-distributed hierarchically MSNs with a pomegranate-like morphology, topped with MIP uniform layer. Electrochemical characterizations were carried out to monitor the properties of the resulting sensing platform based on the MIP/gate effect employing hexacyanoferrate molecules as the electrochemical probe. The data show that due to the high conductivity and electron transfer ability of the prepared theophylline-imprinted membrane, this method exhibits excellent sensitivity and binding affinity with a linear dynamic concentration range in excess of six orders of magnitude and low detection limit (0.66â¯nM), meeting the requirements of theophylline trace analysis.
Assuntos
Impressão Molecular , Nanosferas/química , Polímeros/síntese química , Dióxido de Silício/química , Teofilina/análise , Eletrodos , Limite de Detecção , Polímeros/química , Pirróis/química , Teofilina/químicaRESUMO
Two nine-element sensor arrays, consisting of either three cationic poly(para-phenyleneethynylene)s (PPE) or the same PPEs complexed by cucurbituril[8] (CB[8]) at pH 3, 7, and 13 in water, discriminate 22 different teas and some of their small molecule components, including caffeine, theobromine and theophylline. Both arrays distinguish all of the black, green and oolong teas. The discrimination occurs by differential fluorescence modulation of the components of the sensor array and the treatment of the collected data by linear discriminant analysis. The signal is generated by either simple quenching (PPE only array) or the disruption of the PPE/CB[8] complex and quenching of the complex's or the PPEs' fluorescence through the polyphenolic colorants of the teas. Added amino acids, theobromine, theophylline, and caffeine give a fluorescence turn on of the PPE-CB[8] array, due to the disruption of the self-assembled complex, while for the PPE-alone tongue only caffeine, theobromine, and theophylline elicited useful fluorescence response. Both tongues discriminate different teas without any problem.
Assuntos
Alcinos/química , Técnicas Biossensoriais/métodos , Cafeína/análise , Camellia sinensis/química , Éteres/química , Chá/química , Teobromina/análise , Teofilina/análise , Folhas de Planta/química , Chá/normasRESUMO
In present study, a novel and facile electrochemical sensor based on glassy carbon electrode (GCE) modified with nitrogen-doped carbon nanotubes (N-CNT) decorated with poly (L-Cysteine) (PLCY) were fabricated and applied for the simultaneous voltammetric determination of theophylline (THEO) and caffeine (CAF). The morphology and structure of multilayer film modified on the surface of glassy carbon electrode were investigated successfully by Scanning Electron Microscope (SEM), Transmission Electron Microscope (TEM) and Raman Spectroscopy. And the properties of the modified electrode were investigated by Chronocoulometry (CC), Cyclic Voltammetry (CV) and Differential Pulse Voltammetry (DPV) were utilized to investigate the electrochemical behavior of THEO and CAF on the composite film modified electrode. The results showed that the determination towards THEO and CAF can be operated at the same potential window with the oxidation current peak separated and non-interfering respectively. Compared to the bare GCE, the PLCY/N-CNT/GCE can signally meliorate the electrocatalytic activity towards the oxidation of THEO and CAF with a remarkably increase in the anodic peak currents of 495.94% and 465.48%. Under the optimal conditions, the fabrication multilayer film sensor had excellent performances in determination towards THEO and CAF with a wide linear dynamic range from 0.10 to 70.0µM and 0.40-140.0µM, low detection limit (S/N = 3) of 0.033µM and 0.20µM, respectively. The PLCY/N-CNT/GCE sensor also had advantages as easy-made, high-sensitivity, stability and reproducibility. Moreover, it was successfully used to analyze the THEO and CAF in green tee, oral theophylline sustained release tablets and energy drink sample with a satisfactory result.
Assuntos
Cafeína/análise , Eletroquímica/instrumentação , Limite de Detecção , Nanotubos de Carbono/química , Nitrogênio/química , Peptídeos/química , Teofilina/análise , Cafeína/química , Eletrodos , Concentração de Íons de Hidrogênio , Modelos Moleculares , Conformação Molecular , Teofilina/química , Fatores de TempoRESUMO
Two chromatgraphic methods were developed for determination of Paracetamol (PCM) and Pamabrom (PAM) in presence of P-aminophenol (PAP) and Theophylline (THEO) as potential impurities of both drugs respectively. First method is HPTLC which depends on separation and quantitation of the studied drugs on aluminum plates pre-coated with silica gel 60 F254 as a stationary phase using chloroform:methanol:ethyl acetate:glacial acetic acid (8:0.8:0.6:0.2, v/v/v/v) as mobile phase followed by densitometric measurement of the bands at 254 nm. Second method is RP-HPLC which comprises separation of the studied drugs on a Phenomenex C8 column by gradient elution using mobile phase consisting of sodium dihydrogen phosphate buffer (0.05 M): methanol:acetonitrile (85:10:5, v/v/v) at a flow rate of 1 mL/min for first 7.5 min and (70:20:10, v/v/v) at a flow rate of 1.5 mL/min for the next 5 min. The proposed methods were successfully applied for determination of the potential impurities of PCM and PAM after resolving them from the pure drugs. The developed methods have been validated and proved to meet the requirements delineated by ICH guidelines with respect to linearity, accuracy, precision, specificity and robustness. The validated methods were successfully applied for determination of the studied drugs in their pharmaceutical formulation. The results were statistically compared to those obtained by the reported RP-HPLC method where no significant difference was found; indicating the ability of proposed methods to be used for routine quality control analysis of these drugs.
Assuntos
Acetaminofen/análise , Aminofenóis/análise , Cromatografia Líquida de Alta Pressão/métodos , Contaminação de Medicamentos , Propanolaminas/análise , Teofilina/análogos & derivados , Teofilina/análise , Ácido Acético/química , Acetonitrilas/química , Soluções Tampão , Calibragem , Clorofórmio/química , Combinação de Medicamentos , Concentração de Íons de Hidrogênio , Luz , Metanol/química , Fosfatos/química , Reprodutibilidade dos Testes , Comprimidos/análise , Comprimidos com Revestimento Entérico/análiseRESUMO
RNA probes constitute an important class of functional nucleic acids (FNAs). However, because of their notorious vulnerability to enzymatic degradation, extremely careful and special protocols must be followed when dealing with RNA probes. To fully use the large number of RNA FNAs available for bioanalysis and biomedicine, it is important to explore effective methods to protect RNA probes from enzymatic digestion. In this work, we systematically demonstrate that graphene oxide (GO) can effectively protect RNA probes from enzymatic digestion. Based on this finding, we propose an effective way to design robust RNA biosensors by simply mixing RNA probes with GO for analysis of nucleic acids, proteins, and small molecules. The entire assay is sensitive, selective, rapid, and more importantly, does not require any special protocols. The ability to protect ssRNA from enzymatic digestion by GO offers an exciting new way to stabilize ssRNA, which will not only provide new opportunities to utilize the large number of currently available, yet rarely explored, RNA FNAs for bioanalysis but also offer a new solution to protect important ssRNA molecules, such as microRNA and antisense ssRNA, for a great variety of biomedical applications.
Assuntos
Técnicas Biossensoriais , Grafite/química , Sondas RNA/metabolismo , Aptâmeros de Nucleotídeos/química , Sequência de Bases , DNA/análise , MicroRNAs/química , MicroRNAs/metabolismo , Óxidos/química , Sondas RNA/química , RNA Antissenso/química , RNA Antissenso/metabolismo , Ribonuclease H/metabolismo , Espectrometria de Fluorescência , Teofilina/análise , Fator A de Crescimento do Endotélio Vascular/análiseRESUMO
A novel, simple, accurate and precise RP-HPLC method for simultaneous determination of levosalbutamol sulfate and theophylline has been developed and validated. Separation was achieved on a Phenomenex; C18 column (250 mm × 4.6 mm i.d., 5 µm) using methanol: 10 mM TBAHS(tetrabutyl ammonium hydrogen sulfate) (50:50, v/v) as mobile phase at flow rate of 1.0 mL.min-1. The UV detection wavelength was 274 nm. The linearity is obeyed over a concentration range of 0.5-150 µg.mL-1 with correlation coefficient of 0.999 for both the drugs. The proposed method was validated by determining accuracy, precision, stability and system suitability parameters. The method was found to be robust. Specificity of the method was determined by subjecting the drugs to various stress conditions like acid, alkali, oxidation, thermal and photolytic degradation. The method was used successfully for the simultaneous determination of levosalbutamol sulfate and theophylline in syrup dosage form.
Desenvolveu-se e validou-se método de RP-HPLC novo, simples, exato e preciso de determinação simultânea do sulfato de levossalbutamol e teofilina.. A separação foi efetuada em uma coluna Phenomenex; C18 (250 mm x 4,6 mm d.i., 5 µm) utilizando metanol: TBAHS (hidrogenossulfato de tetrabutilamônio) 10 mM (50:50, v/v) como fase móvel, com fluxo de 1,0 mL.min-1. O comprimento de onda de detecção no UV foi 274 nm. Observou-se linearidade na faixa de concentração de 0,5-150 µg mL-1, com coeficiente de correlação de 0,999 para ambos os fármacos. O método proposto foi validado determinando-se exatidão, precisão, estabilidade e parâmetros de adequação do sistema. O método mostrou-se robusto. A especificidade do método foi determinada submetendo os fármacos a várias condições de estresse, como ácido, álcali, oxidação, degradação térmica e fotolítica. O método foi usado com sucesso para a determinação simultânea do sulfato de levossalbutamol e teofilina na forma de xarope.
Assuntos
Teofilina/análise , Cromatografia Líquida de Alta Pressão/métodos , Levalbuterol/análise , Formas de DosagemRESUMO
The purpose of the present study was to investigate the interaction between ketotifen fumarate and anhydrous theophylline in aqueous media of various pH (1.2 and 6.8). Using Job's continuous-variation analysis and Ardon's spectrophotomeric measurement methods, the values of the stability constants of theophylline with ketotifen were determined at a fixed temperature (37 ºC) at various pH. The stability constants, ranging between 5.66 and 9.92, were derived from Ardon's plot, indicating that comparatively stable complexes had formed as a result of an interaction between the drugs. However, following the interaction of theophylline with ketotifen, stability constants were <1 at gastric pH (1.2) and intestinal pH (6.8). Concurrent administration of ketotifen and theophylline could result in the formation of a stable complex and this is likely to reduce the therapeutic activities of both drugs.
O objetivo do presente estudo foi investigar a interação entre o fumarato de cetotifeno e a teofilina anidra em meios aquosos com vários pH (1,2 e 6,8). Utilizando a análise da variação contínua de Job e os métodos de medida espectrofotométrica de Ardon, os valores das constantes de estabilidade da teofilina com o cetotifeno foram determinados em temperatura fixa (37 oC) em vários pH. As constantes de estabilidade, variando entre 5,66 e 9,92 derivaram-se a partir do delineamento de Ardon, indicando, comparativamente, que complexos estáveis se formaram como resultado da interação entre os fármacos. Entretanto, seguindo a interação da teofilina com o cetotifeno, as constantes de estabilidade foram <1, em pH gástrico (1,2) e intestinal (8,8). A administração concomitante de cetotifeno e teofilina poderia resultar na formação de complexo estável, o que reduz a atividade terapêutica de ambos os fármacos.
Assuntos
Técnicas In Vitro/métodos , Cetotifeno/análise , Teofilina/análise , Reatividade-EstabilidadeRESUMO
This work aimed to evaluate the phytochemical content and to determine the antioxidant and cytotoxic activities of methanol extracts of the carob tree (Ceratonia siliqua L.) germ flour. The extracts were rich in phenolic compounds, had considerable antioxidant activity, and reduced the viability of cervical (HeLa) cancer cells. The chemical content and the biological activities of the extracts were significantly affected by gender and cultivar. Female cultivar Galhosa had the highest levels of phenolic compounds, and the highest antioxidant activity. Extracts from the hermaphrodite trees and from the female cultivars Galhosa and Costela/Canela exhibited the highest cytotoxic activity. The most abundant compound was theophylline. The phenolic content was correlated to both antioxidant and cytotoxic activities. Our findings provide new knowledge about the health implications of consuming food supplemented with carob germ flour.
Assuntos
Antioxidantes/farmacologia , Fabaceae/química , Galactanos/farmacologia , Mananas/farmacologia , Extratos Vegetais/farmacologia , Gomas Vegetais/farmacologia , Teofilina/farmacologia , Galactanos/química , Células HeLa , Humanos , Mananas/química , Fenóis/análise , Fenóis/farmacologia , Extratos Vegetais/análise , Gomas Vegetais/química , Teofilina/análiseRESUMO
This article presents some experience obtained by applying capillary gas chromatography coupled with thermal conductivity detection (GC/TCD) to the determination of water in substances for pharmaceutical use. This technique represents a useful, orthogonal tool complementary to water determination methods based on volumetric or coulometric titration. It can also represent an alternative technique when such titrations are not applicable. This article presents the preliminary results obtained in a number of case studies where a GC/TCD procedure was applied in comparison with pharmacopoeial methods to substances with different water contents.
Assuntos
Preparações Farmacêuticas/análise , Água/análise , Amoxicilina/análise , Antibacterianos/análise , Cromatografia Gasosa/métodos , Ciprofloxacina/análogos & derivados , Ciprofloxacina/análise , União Europeia , Indicadores e Reagentes , Solventes , Espectinomicina/análise , Teofilina/análise , Condutividade TérmicaRESUMO
BACKGROUND: Recently, we identified hepatocytes as the major cellular source of profibrogenic connective tissue growth factor (CTGF/CCN2) in the liver. Based on reports of a hepatoprotective effect of coffee consumption, we were the first to provide evidence that caffeine suppresses transforming growth factor (TGF)-beta dependent and -independent CTGF expression in hepatocytes in vitro and in vivo, thus suggesting this xanthine-alkaloid as a potential therapeutic agent. AIM: This study aims at comparing the inhibitory capacities of caffeine and its three demethylated derivates paraxanthine, theophylline and theobromine on CTGF expression in hepatocytes and hepatic stellate cells (HSC). RESULTS: Our data suggest paraxanthine as the most important pharmacological repressor of hepatocellular CTGF expression among the caffeine-derived metabolic methylxanthines with an inhibitory dosage (ID)50 of 1.15 mM, i.e. 3.84-fold lower than what is observed for caffeine. In addition, paraxanthine displayed the least cell toxicity as proven by the water-soluble tetrazolium-1 cell vitality assay. However, caffeine or any of the metabolites did not inhibit CTGF expression in HSC. At the toxicological threshold concentration of 1 mM for paraxanthine, we observed an inhibition of hepatocellular CTGF synthesis by 44%, which was strongly reverted in the presence of the specific competitive cyclic adenosine monophosphate inhibitor Rp-adenosine 3',5-cyclic monophosphorothioate triethylammonium salt. Furthermore, CTGF protein expression induced by various concentrations of TGF-beta (0.13-1 ng/ml) is still reduced by, on average, 27%/45% in the presence of paraxanthine (1.25 mM/2.5 mM). CONCLUSION: Our data provide an evidence-based suggestion of the caffeine-derived primary metabolite paraxanthine as a potentially powerful antifibrotic drug by its inhibitory effect on (hepatocellular) CTGF synthesis.
Assuntos
Cafeína/química , Fator de Crescimento do Tecido Conjuntivo/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Hepatócitos/metabolismo , Teofilina/farmacologia , Animais , Western Blotting , Células Cultivadas , Primers do DNA/genética , Eletroforese em Gel de Poliacrilamida , Concentração Inibidora 50 , Masculino , Estrutura Molecular , Ratos , Ratos Sprague-Dawley , Análise de Regressão , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Teobromina/farmacologia , Teofilina/análiseRESUMO
We screened a series of RNA and DNA aptamers for their ability to serve in the dye displacement assays in which analytes compete with TO dye. We conclude that, while the performance of the TO dye displacement approach is not always predictable, it is still a simple and sensitive assay to detect binding between RNA aptamers and small molecules. In particular, we describe efficient assays for tobramycin and theophylline, with up to 90% displacement of TO observed, and we describe the first aptameric assay for cAMP.
Assuntos
Aptâmeros de Nucleotídeos/química , Benzotiazóis/química , Corantes Fluorescentes/química , Quinolinas/química , Trifosfato de Adenosina/análise , AMP Cíclico/análise , Guanosina Trifosfato/análise , Conformação de Ácido Nucleico , Sensibilidade e Especificidade , Teofilina/análise , Tobramicina/análiseRESUMO
We report the first examples of modular aptameric sensors, which transduce recognition events into fluorescence changes through allosteric regulation of noncovalent interactions with a fluorophore. These sensors consist of: (a) a reporting domain, which signals the binding event of an analyte through binding to a fluorophore; (b) a recognition domain, which binds the analyte; and (c) a communication module, which serves as a conduit between recognition and signaling domains. We tested recognition regions specific for ATP, FMN, and theophylline in combinations with malachite green binding aptamer as a signaling domain. In each case, we were able to obtain a functional sensor capable of responding to an increase in analyte concentration with an increase in fluorescence. Similar constructs that consist only of natural RNA could be expressed in cells and used as sensors for intracellular imaging.
Assuntos
Técnicas Biossensoriais/métodos , Oligonucleotídeos/química , Trifosfato de Adenosina/análise , Trifosfato de Adenosina/química , Sequência de Bases , Corantes/química , DNA/química , Mononucleotídeo de Flavina/análise , Corantes Fluorescentes/química , Conformação de Ácido Nucleico , Oligonucleotídeos/síntese química , Oligonucleotídeos/metabolismo , Corantes de Rosanilina/química , Espectrometria de Fluorescência , Teofilina/análiseRESUMO
The influence of different factors on the formation of nanoparticles in freshly brewed tea extracts was investigated. A black tea infusion was observed during cooling using photon correlation spectroscopy (PCS). The mean particle size and the number of the nanoparticles increase with decreasing temperature. In the presence of caffeine more particles are formed within the infusion. To study the influence of slight structural differences between methylxanthines, the effect of the addition of caffeine to solutions of freshly prepared decaffeinated tea was compared to that of theophylline and theobromine. In the case of theophylline fewer nanoparticles were formed. Molecular modelling calculations were performed to evaluate the most probable geometries for caffeine-polyphenol complexes. A parallel position and a congruent orientation of the 6-membered ring of caffeine and the aromatic galloyl group is the most probable geometry.
Assuntos
Chá/química , Absorciometria de Fóton , Cafeína/análise , Microesferas , Modelos Moleculares , Tamanho da Partícula , Extratos Vegetais/análise , Teobromina/análise , Teofilina/análiseRESUMO
BACKGROUND: Ischemic preconditioning is characterized by the limitation of infarct size or ischemic signs after one or more brief episodes of ischemia, a process that probably involves stimulation of adenosine receptors. One human model of ischemic preconditioning is repetitive occlusion of a coronary artery during angioplasty. By using this method of inducing ischemia, we tested the hypothesis that blockade of adenosine receptors with aminophylline would abolish ischemic preconditioning in human beings. METHODS: Twenty-six patients undergoing angioplasty were randomly assigned to receive either aminophylline (6 mg/kg IV) or placebo before repetitive coronary occlusion (two 2-minute occlusions separated by 5 minutes). ST-segment changes on the surface electrocardiogram were used as a measure of myocardial ischemia. Serum theophylline levels and the conduction response to an intravenous bolus of adenosine were used to assess the efficacy of adenosine receptor blockade. RESULTS: Repetitive coronary occlusion resulted in a reduction in ST-segment shift in 9 of 13 patients given placebo. In contrast, 9 of 13 patients receiving aminophylline had an increase in ST-segment shift on the second occlusion (P =.002). Patients receiving aminophylline (mean serum theophylline level of 8.38 +/- 0.45 mg/dL) did not have significant conduction block with intravenous adenosine. CONCLUSIONS: Repetitive coronary occlusion reduces the signs of ischemia in human beings, a process limited by blockade of adenosine receptors.
Assuntos
Aminofilina/farmacologia , Cardiotônicos/farmacologia , Doença das Coronárias/patologia , Precondicionamento Isquêmico Miocárdico , Isquemia Miocárdica/fisiopatologia , Receptores Purinérgicos P1/fisiologia , Angioplastia , Eletrocardiografia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Antagonistas de Receptores Purinérgicos P1 , Teofilina/análiseRESUMO
A new method for a comprehensive screening and confirmation of beta-2 agonists in human urine is presented based on gas chromatography-low-resolution mass spectrometry (GC-MS) using electron impact ionisation (EI). After hydrolysis of the conjugates with beta-glucuronidase/arylsulfatase a derivatisation step with formaldehyde converts fenoterol, orciprenaline, reproterol and terbutaline to one derivative, a tetrahydroisoquinoline, while the other beta-2 agonists remain unchanged. Liquid-liquid extraction and trimethylsilylation follow. The tetrahydroisoquinoline derivatives show good gas chromatographic and mass spectrometric behaviour. The detection limit of these four beta-2 agonists in the screening using low-resolution mass spectrometry is 10 ng/ml of urine. The other beta-2 agonists are detected as parent compounds with the same recovery after sample preparation with and without formaldehyde. The EI mass spectra of the tetrahydroisoquinoline derivatives are presented.