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Biosensors based on ion-sensitive field effect transistors (ISFETs) combined with aptamers offer a promising and convenient solution for point-of-care testing applications due to the ability for fast and label-free detection of a wide range of biomarkers. Mobile and easy-to-use readout devices for the ISFET aptasensors would contribute to further development of the field. In this paper, the development of a portable PC-controlled device for detecting aptamer-target interactions using ISFETs is described. The device assembly allows selective modification of individual ISFETs with different oligonucleotides. Ta2O5-gated ISFET structures were optimized to minimize trapped charge and capacitive attenuation. Integrated CMOS readout circuits with linear transfer function were used to minimize the distortion of the original ISFET signal. An external analog signal digitizer with constant voltage and superimposed high-frequency sine wave reference voltage capabilities was designed to increase sensitivity when reading ISFET signals. The device performance was demonstrated with the aptamer-driven detection of troponin I in both reference voltage setting modes. The sine wave reference voltage measurement method reduced the level of drift over time and enabled a lowering of the minimum detectable analyte concentration. In this mode (constant voltage 2.4 V and 10 kHz 0.1Vp-p), the device allowed the detection of troponin I with a limit of detection of 3.27 ng/mL. Discrimination of acute myocardial infarction was demonstrated with the developed device. The ISFET device provides a platform for the multiplexed detection of different biomarkers in point-of-care testing.
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Aptámeros de Nucleótidos , Biomarcadores , Técnicas Biosensibles , Transistores Electrónicos , Aptámeros de Nucleótidos/química , Técnicas Biosensibles/instrumentación , Técnicas Biosensibles/métodos , Biomarcadores/análisis , Humanos , Troponina I/análisis , Troponina I/sangreRESUMEN
Fabry disease is a lysosomal storage disorder caused by a significant decrease in the activity or absence of the enzyme α-galactosidase A. The diagnostics of Fabry disease during newborn screening are reasonable, due to the availability of enzyme replacement therapy. This paper presents an electrochemical method using complementary metal-oxide semiconductor (CMOS)-compatible ion-sensitive field effect transistors (ISFETs) with hafnium oxide-sensitive surfaces for the detection of α-galactosidase A activity in dried blood spot extracts. The capability of ISFETs to detect the reaction catalyzed by α-galactosidase A was demonstrated. The buffer composition was optimized to provide suitable conditions for both enzyme and ISFET performance. The use of ISFET structures as sensor elements allowed for the label-free detection of enzymatic reactions with melibiose, a natural substrate of α-galactosidase A, instead of a synthetic fluorogenic one. ISFET chips were packaged with printed circuit boards and microfluidic reaction chambers to enable long-term signal measurement using a custom device. The packaged sensors were demonstrated to discriminate between normal and inhibited GLA activity in dried blood spots extracts. The described method offers a promising solution for increasing the widespread distribution of newborn screening of Fabry disease.
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Técnicas Biosensibles , Pruebas con Sangre Seca , Enfermedad de Fabry , Transistores Electrónicos , alfa-Galactosidasa , alfa-Galactosidasa/sangre , Pruebas con Sangre Seca/métodos , Humanos , Enfermedad de Fabry/sangre , Enfermedad de Fabry/diagnóstico , Técnicas Biosensibles/métodos , Técnicas Biosensibles/instrumentación , Recién Nacido , Tamizaje Neonatal/métodosRESUMEN
The vertebral column is a hallmark of vertebrates; it is the structural basis of their body and the locomotor apparatus in particular. Locomotion of any vertebrate animal in its typical habitat is directly associated with functional adaptations of its vertebrae. This study is the first large-scale analysis of mobility throughout the presacral region of the vertebral column covering a majority of extant odd-toed ungulates from 6 genera and 15 species. In this study, we used a previously developed osteometry-based method to calculate available range of motion. We quantified all three directions of intervertebral mobility: sagittal bending (SB), lateral bending (LB), and axial rotation (AR). The cervical region in perissodactyls was found to be the most mobile region of the presacral vertebral column in LB and SB. Rhinoceroses and tapirs are characterized by the least mobile necks in SB among odd-toed and even-toed ungulates. Equidae are characterized by very mobile necks, especially in LB. The first intrathoracic joint (T1-T2) in Equidae and Tapiridae is characterized by significantly increased mobility in the sagittal plane compared to the typical thoracic joints and is only slightly less mobile than typical cervical joints. The thoracolumbar part of the vertebral column in odd-toed ungulates is very stiff. Perissodactyls are characterized by frequent fusions of vertebrae with each other with complete loss of mobility. The posterior half of the thoracic region in perissodactyls is characterized by especially stiff intervertebral joints in the SB direction. This is probably associated with hindgut fermentation in perissodactyls: the sagittal stiffness of the posterior thoracic region of the vertebral column is able to passively support the hindgut heavily loaded with roughage. Horses are known as a prime example of a dorsostable galloper among mammals. However, based on SB in the lumbosacral part of the backbone, equids appear to be the least dorsostable among extant perissodactyls; the cumulative SB in equids and tapirs is as low as in the largest representatives of artiodactyls, while in Rhinocerotidae it is even lower representing the minimum across all odd-toed and even-toed ungulates. Morphological features of small Paleogene ancestors of rhinoceroses and equids indicate that dorsostability is a derived feature of perissodactyls and evolved convergently in the three extant families.
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Perisodáctilos , Columna Vertebral , Caballos , Animales , Equidae , Mamíferos , Locomoción , Fenómenos BiomecánicosRESUMEN
In this study, growth parameters of underground parts and concentrations of phenylpropanoids, phenylethanoids, flavonoids, hydroxybenzoic acids, and catechins in aqueous-ethanol extracts of 6-year-old cultivated plants of Rhodiola rosea (propagated in vitro) of Altai Mountain origin were analyzed, and differences in chemical composition among plant specimens and between plant parts (rhizome and root) were evaluated. High-performance liquid chromatography detected 13 phenolic compounds. Roots contained 1.28 times higher phenylethanoids levels (1273.72 mg/100 g) than rhizomes did. Overall, the concentration of phenylethanoids in underground organs was not high and ranged from 21.36 to 103.00 mg/100 g. High variation among R. rosea individual plants was noted both in growth characteristics and in levels of secondary metabolites under our cultivation conditions. It was found that concentrations of phenylpropanoids, phenylethanoids, and catechins significantly depend on the plant part analyzed (p ≤ 0.05). Specimen No. 4 is characterized by the highest concentration of rosavins (1230.99 mg/plant) and the lowest concentration of cinnamyl alcohol (62.87 mg/plant). Despite the wide range of values, all 10 tested specimens (underground part) met the minimum requirements of the United States Pharmacopeia (2015) for rosavins (0.3%) and of the Russia State Pharmacopoeia (2015) for the average level of rosavins (roots): (1%).
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Catequina , Rhodiola , Raíces de Plantas/química , Extractos Vegetales/química , Rhodiola/química , Siberia , IndividualidadRESUMEN
Copper-64 (T1/2 = 12.7 h) is a positron and beta-emitting isotope, with decay characteristics suitable for both positron emission tomography (PET) imaging and radiotherapy of cancer. Copper-67 (T1/2 = 61.8 h) is a beta and gamma emitter, appropriate for radiotherapy ß-energy and with a half-life suitable for single-photon emission computed tomography (SPECT) imaging. The chemical identities of 64Cu and 67Cu isotopes allow for convenient use of the same chelating molecules for sequential PET imaging and radiotherapy. A recent breakthrough in 67Cu production opened previously unavailable opportunities for a reliable source of 67Cu with high specific activity and purity. These new opportunities have reignited interest in the use of copper-containing radiopharmaceuticals for the therapy, diagnosis, and theranostics of various diseases. Herein, we summarize recent (2018-2023) advances in the use of copper-based radiopharmaceuticals for PET, SPECT imaging, radiotherapy, and radioimmunotherapy.
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Neoplasias , Radiofármacos , Humanos , Radiofármacos/química , Cobre , Tomografía de Emisión de Positrones/métodos , Tomografía Computarizada de Emisión de Fotón Único , Neoplasias/diagnóstico por imagen , Neoplasias/radioterapiaRESUMEN
The paper focuses on the growth dynamics and biosynthetic characteristics of the microshoot culture of Spiraea betulifolia ssp. aemiliana obtained in vitro in agar-solidified and liquid media. Microshoots cultured in either type of media showed similar growth dynamics. The most active culture growth was observed from day 35 to day 60. A comparative analysis of the contents of flavonoids and phenol carboxylic acids showed a higher level of phenol carboxylic acids (5.3-6.84%) and a stronger 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging activity (half-maximal inhibitory concentration: 341 µg/mL) in S. betulifolia ssp. aemiliana microshoots grown in the liquid medium compared to the microshoots cultured in the solid medium. The flavonoid content of the cultured microshoot did not depend on the consistency of the medium. High-performance liquid chromatography (HPLC) was employed to study the profile and levels of phenolic compounds in microshoots, intact plants, and ex vitro-acclimated S. betulifolia ssp. aemiliana plants. The concentration of kaempferol glycosides was found to be higher in microshoots (1.33% in the solid medium, 1.06% in the liquid medium) compared to intact plants and ex vitro-acclimated plants. Thus, the microshoots of S. betulifolia ssp. aemiliana cultured in the liquid medium rapidly increase their biomass and are an inexpensive promising source of biologically active antioxidant substances, mainly phenol carboxylic acids and kaempferol glycosides.
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Quempferoles , Spiraea , Quempferoles/análisis , Flavonoides/farmacología , Antioxidantes/farmacología , Extractos Vegetales/farmacología , Fenoles/análisis , Glicósidos , Ácidos Carboxílicos , Cromatografía Líquida de Alta PresiónRESUMEN
This study is the first analysis of mobility in the lumbosacral joint of even-toed ungulates covering the full range of body masses and running forms. In this study, we modified a previously developed osteometry-based method to calculate the available range of motion (aROM) in the lumbosacral joint in artiodactyls. We quantified all three directions of intervertebral mobility: sagittal bending (SB), lateral bending (LB), and axial rotation (AR). This research covers extant artiodactyls from 10 families, 57 genera, and 78 species. The lumbosacral joint in artiodactyls is on average almost twice as mobile in SB as the average intralumbar joint (aROM 15.68° vs 8.22°). In all artiodactyls, the first sacral prezygapophyses are equipped with postfacet fossae determining the available range of lumbosacral hyperextension. SB aROM in the lumbosacral joint in artiodactyls varies almost sevenfold (from 4.53° to 31.19°) and is closely related to the body mass and running form. An allometric equation was developed for the first time, for the joint angular amplitude of motion, exemplified by the artiodactyl lumbosacral SB aROMs, as a power function of body mass, the power coefficient value being close to -0.15. High SB aROM at the lumbosacral joint is characteristic of artiodactyls with at least one of the following characteristics: high cumulative and average SB aROM in the lumbar region (Pearson r = 0.467-0.617), small body mass (r = -0.531), saltatorial or saltatorial-cursorial running form (mean = 16.91-18.63°). The highest SB aROM in the lumbosacral joint is typical for small antelopes and Moschidae (mean = 20.24-20.27°). Among these artiodactyls SB aROMs in the lumbosacral joint are on par with various carnivores. Large and robust artiodactyls, adapted predominantly to mediportal and stilt (running on extremely tall limbs) running forms, have 2-3 times smaller SB aROMs in the lumbosacral joint. Adaptation to endurance galloping in open landscapes (cursorial running form) is accompanied by smaller lumbar and lumbosacral SB aROMs compared to that in saltatorial-cursorial artiodactyls of the same body mass. The wide range of species studied makes it possible to significantly expand the knowledge of relations of the mobility of the lumbosacral joint in artiodactyls to body mass and running form.
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Región Lumbosacra , Pelvis , Fenómenos Biomecánicos , Humanos , Vértebras Lumbares , Rango del Movimiento Articular , SacroRESUMEN
An electrochemical aptamer-based sensor was developed for glutamate, the major excitatory neurotransmitter in the central nervous system. Determining glutamic acid release and glutamic acid levels is crucial for studying signal transmission and for diagnosing pathological conditions in the brain. Glutamic acid-selective oligonucleotides were isolated from an ssDNA library using the Capture-SELEX protocol in complex medium. The selection permitted the isolation of an aptamer 1d04 with a dissociation constant of 12 µM. The aptamer sequence was further used in the development of an electrochemical aptamer sensor. For this purpose, a truncated aptamer sequence named glu1 was labelled with a ferrocene redox tag at the 3'-end and immobilized on a gold electrode surface via Au-thiol bonds. Using 6-mercapto-1-hexanol as the backfill, the sensor performance was characterized by alternating current voltammetry. The glu1 aptasensor showed a limit of detection of 0.0013 pM, a wide detection range between 0.01 pM and 1 nM, and good selectivity for glutamate in tenfold diluted human serum. With this enzyme-free aptasensor, the highly selective and sensitive detection of glutamate was demonstrated, which possesses great potential for implementation in microelectrodes and for in vitro as well as in vivo monitoring of neurotransmitter release.
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Aptámeros de Nucleótidos/química , Técnicas Electroquímicas/métodos , Ácido Glutámico/sangre , Técnicas Biosensibles/métodos , Ácido Glutámico/análisis , Hexanoles/química , Humanos , Límite de Detección , Compuestos de Sulfhidrilo/químicaRESUMEN
In our work, the photonuclear production of 198,199Au isotopes for nuclear medicine purposes was studied, and a method for their recovery from irradiated mercury was developed. The yields of the corresponding nuclear reactions were determined, and a comparison of various methods of obtaining gold radioisotopes was provided. New sorbents based on benzo-15-crown-5, which selectively binds gold, were studied, and the optimal conditions for Au recovery with a high degree of purification from mercury were found. It was established that, for the fast and quantitative recovery of Au isotopes, it was necessary to add at least 0.1 mg of the carrier. As a result, the developed method can be regularly used to obtain 198,199Au for the research of radiopharmaceuticals based on them.
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Alquimia , Mercurio , Electrones , Oro , IsótoposRESUMEN
A new method of production of one of the most widely used isotopes in nuclear medicine, 177Lu, with high chemical purity was developed; this method includes irradiation of the HfO2 target with bremsstrahlung photons. The irradiated target was dissolved in HF and then diluted and placed onto a column filled with LN resin. Quantitative sorption of 177Lu could be observed during this process. The column later was rinsed with the mixture of 0.1 M HF and 1 M HNO3 and then 2 M HNO3 to remove impurities. Quantitative desorption of 177Lu was achieved by using 6 M HNO3. The developed method of 177Lu production ensures high purification of this isotope from macroquantities of hafnium and zirconium and radioactive impurities of carrier-free yttrium. The content of 177mLu in 177Lu in photonuclear production was determined. Due to high chemical and radionuclide purity, 177Lu obtained by the developed method can be used in nuclear medicine.
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Medicina Nuclear , Hafnio , Lutecio/química , Radioisótopos/química , CintigrafíaRESUMEN
Nowadays, cobalt isotopes 55Co, 57Co, and 58mCo are considered to be promising radionuclides in nuclear medicine, with 55Co receiving the most attention as an isotope for diagnostics by positron emission tomography. One of the current research directions is dedicated to its production using electron accelerators (via photonuclear method). In our work, the yields of nuclear reactions occurring during the irradiation of natNi and 60Ni by bremsstrahlung photons with energy up to 55 MeV were determined. A method of fast and simple cobalt isotopes separation from irradiated targets using extraction chromatography was developed.
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In this paper, we develop and validate an osteometry-based mechanistic approach to calculation of available range of motion (aROM) in presacral intervertebral joints in sagittal bending (SB), lateral bending (LB), and axial rotation (AR). Our basic assumption was the existence of a mechanistic interrelation between the geometry of zygapophysial articular facets and aROM. Trigonometric formulae are developed for aROM calculation, of which the general principle is that the angle of rotation is given by the ratio of the arc length of motion to the radius of this arc. We tested a number of alternative formulae against available in vitro data to identify the most suitable geometric ratios and coefficients for accurate calculation. aROM values calculated with the developed formulae show significant correlation with in vitro data in SB, LB, and AR (Pearson r = 0.900) in the reference mammals (man, sheep, pig, cow). It was found that separate formulae for different zygapophysial facet types (radial (Rf), tangential (Tf), radial with a lock (RfL)) give significantly greater accuracy in aROM calculation than the formulae for the presacral spine as a whole and greater accuracy than the separate formulae for different spine regions (cervical, thoracic, lumbar). The advantage of the facet-specific formulae over the region-specific ones shows that the facet type is a more reliable indicator of the spine mobility than the presence or absence of ribs. The greatest gain in calculation accuracy with the facet-specific formulae is characteristic in AR aROM. The most important theoretical outcome is that the evolutionary differentiation of the zygapophysial facets in mammals, that is the emergence of Tf joints in the rib cage area of the spine, was more likely associated with the development of AR rather than with SB mobility and, hence, with cornering rather than with forward galloping. The AR aROM can be calculated with the formulae common for man, sheep, pig, and cow. However, the SB aROM of the human spine is best calculated with different coefficient values in the formulae than those for studied artiodactyls. The most suitable coefficient values indicate that the zygapophysial articular facets tend to slide past each other to a greater extent in the human thoracolumbar spine rather than in artiodactyls. Due to this, artiodactyls retain relatively greater facet overlap in extremely flexed and extremely extended spine positions, which may be more crucial for their quadrupedal gallop than for human bipedal locomotion. The SB, LB, and AR aROMs are quite separate in respect of the formulae structure in the cervical region (radial facet type). However, throughout the thoracolumbar spine (tangential and radial with lock facets), the formulae for LB and AR are basically similar differing in coefficient values only. This means that, in the thoracolumbar spine, the greater the LB aROM, the greater the AR aROM, and vice versa. The approach developed promises a wide osteological screening of extant and extinct mammals to study the sex, age, geographical variations, and disorders.
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Disco Intervertebral/fisiología , Rango del Movimiento Articular/fisiología , Columna Vertebral/fisiología , Articulación Cigapofisaria/fisiología , Animales , Bovinos , Humanos , Rotación , Ovinos , Porcinos , Soporte de Peso/fisiologíaRESUMEN
In this study, we used a previously developed osteometry-based method to calculate available range of motion in presacral intervertebral joints in artiodactyls. We have quantified all three directions of intervertebral mobility: sagittal bending (SB), lateral bending (LB), and axial rotation (AR). This research covers 10 extant families of artiodactyls from 33 genera and 39 species. The cervical region in artiodactyls is the most mobile region of the presacral vertebral column in SB and LB. Mobility is unevenly distributed throughout the joints of the neck. The posterior neck joints (C4-C7) are significantly more mobile (on average by 2.5-3.5°) to anterior joints (C2-C4) and to the neck-thorax joint (C7-T1) in SB and LB. An increase in the relative length of the cervical region in artiodactyls is accompanied by an increase in the bending amplitudes (SB: Pearson r = 0.781; LB: r = 0.884). Animals with the most mobile necks (representative of Giraffidae and Camelidae) are 2-3 times more mobile in SB and LB compared to species with the least mobile necks. The thoracic region in artiodactyls, as in other mammals, is characterized by the greatest amplitudes of AR due to the tangential orientation of the zygapophyseal articular facets. The lowest AR values in the thoracic region are typical for the heaviest artiodactyls-Hippopotamidae. The highest AR values are typical for such agile runners as cervids, musk deer, pronghorn, as well as large and small antelopes. SB mobility in the posterior part of the thoracic region can be used by artiodactyls during galloping. The highest values of SB aROM in the posterior part of the thoracic region are typical for small animals with high SB mobility in the lumbar region. The lumbar region in mammals is adapted for efficient SB. Both the cumulative and average SB values in the lumbar region showed correspondence to the running type employed by an artiodactyl. The greatest SB amplitudes in the lumbar region are typical for small animals, which use saltatorial and saltatorial-cursorial running. An increase in body size also corresponds to a decrease in lumbar SB amplitudes. The lowest SB amplitudes are typical for species using the so-called mediportal running. Adaptation to endurance galloping in open landscapes is accompanied by a decrease in lumbar SB amplitudes in artiodactyls. The consistency of the approach used and the wide coverage of the studied species make it possible to significantly expand and generalize the knowledge of the biomechanics of the vertebral column in artiodactyls.
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Ciervos , Articulación Cigapofisaria , Animales , Fenómenos Biomecánicos , Vértebras Lumbares , Rango del Movimiento Articular , Columna VertebralRESUMEN
Aptamers are nucleic acid ligands that bind specifically to a target of interest. Aptamers have gained in popularity due to their high potential for different applications in analysis, diagnostics, and therapeutics. The procedure called systematic evolution of ligands by exponential enrichment (SELEX) is used for aptamer isolation from large nucleic acid combinatorial libraries. The huge number of unique sequences implemented in the in vitro evolution in the SELEX process imposes the necessity of performing extensive sequencing of the selected nucleic acid pools. High-throughput sequencing (HTS) meets this demand of SELEX. Analysis of the data obtained from sequencing of the libraries produced during and after aptamer isolation provides an informative basis for precise aptamer identification and for examining the structure and function of nucleic acid ligands. This review discusses the technical aspects and the potential of the integration of HTS with SELEX.
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Aptámeros de Nucleótidos/análisis , Biblioteca de Genes , Secuenciación de Nucleótidos de Alto Rendimiento/métodos , Ácidos Nucleicos/genética , Técnica SELEX de Producción de Aptámeros/instrumentación , Aptámeros de Nucleótidos/síntesis química , Aptámeros de Nucleótidos/genética , Secuencia de Bases , Benchmarking , Secuenciación de Nucleótidos de Alto Rendimiento/instrumentación , Humanos , Ligandos , Conformación de Ácido Nucleico , Ácidos Nucleicos/química , Ácidos Nucleicos/metabolismo , Medicina de Precisión/métodosRESUMEN
Aptamers are small oligonucleotides that are capable of binding specifically to a target, with impressive potential for analysis, diagnostics, and therapeutics applications. Aptamers are isolated from large nucleic acid combinatorial libraries using an iterative selection process called SELEX (Systematic Evolution of Ligands by EXponential enrichment). Since being implemented 30 years ago, the SELEX protocol has undergone many modifications and improvements, but it remains a laborious, time-consuming, and costly method, and the results are not always successful. Each step in the aptamer selection protocol can influence its results. This review discusses key technical points of the SELEX procedure and their influence on the outcome of aptamer selection.
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Aptámeros de Nucleótidos/química , Cartilla de ADN/química , Técnica SELEX de Producción de Aptámeros/métodos , Aptámeros de Nucleótidos/síntesis química , Cartilla de ADN/síntesis química , ADN de Cadena Simple/aislamiento & purificación , Biblioteca de Genes , Secuenciación de Nucleótidos de Alto Rendimiento/métodos , Técnicas de Amplificación de Ácido Nucleico , Ácidos Nucleicos/química , Reacción en Cadena de la PolimerasaRESUMEN
Furaneol is an aroma compound which occurs naturally in foods and is used as an artificial flavor. Detection of furaneol is required in food science and food processing industry. Capture- Systematic Evolution of Ligands by EXponential enrichment (SELEX) protocol was applied for the isolation of an aptamer binding to furaneol, a small volatile organic substance contributing to the flavor of various products. Thirteen cycles of selection were performed. The resulting DNA pool was cloned, using blunt-end cloning, and ninety-six plasmids were sequenced and analyzed. Eight oligonucleotides were selected as aptamer candidates and screened for the ability to bind to furaneol, using three different methods-magnetic-beads associated elution assay, SYBR Green I assay, and exonuclease protection assay. One of the candidates was further characterized as an aptamer. The apparent equilibrium constant was determined to be (1.1 ± 0.4) µM, by the fluorescent method. The reported aptamer was applied for development of the ion-sensitive field-effect transistor (ISFET)-based biosensor, for the analysis of furaneol, in the concentration range of 0.1â»10 µM.
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Aptámeros de Nucleótidos/química , ADN de Cadena Simple/química , Furanos/química , Técnica SELEX de Producción de Aptámeros/métodos , Aptámeros de Nucleótidos/genética , Secuencia de Bases , Técnicas Biosensibles , FluorescenciaRESUMEN
An aptamer for vanillin was obtained and then used for the development of an aptasensor based on an ion-sensitive field-effect transistor (ISFET). This aptamer (a single-stranded DNA;ssDNA) was selected using the Capture-SELEX protocol, which suites well for selection of aptamers to small molecules. Among six aptamer candidates, the aptamer Van_74 with the highest affinity for vanillin was chosen (elution of 35% of the aptamer from a solid support in the presence of 2 mM of vanillin). Van_74 was characterized using nondenaturating PAGE of washouts from magnetic beads. It is shown that Van_74 binds to vanillin with an dissociation constant of >7.8 µM (determined by nondenaturating PAGE) and it was specific to vanillin in comparison with interferents: benzaldehyde, guaiacol, furaneol, ethyl guaiacol and ethyl vanillin. Also it was shown that change of buffer composition greatly affected the binding ability of Van_74. For biosensor fabrication aptamer was immobilised on the Ta2O5-sensitive surface of the ISFET via "click-chemistry". Detection scheme implied dehybridisation of the ssDNA probe from the aptamer and release in the solution during the addition of vanillin. As a result, the surface potential increase upon vanillin binding with the aptamer was detected by the transistor. The biosensor had a detection limit of 1.55 × 10-7 M and a dynamic range from 1.55 × 10-7 M to 1 × 10-6 M. Effective constant Kd,eff for vanillin binding on biosensor surface was calculated to be (9 ± 3) × 10-7 M. This allows selective detection of vanillin in the mixture of interferents and in samples of coffee extract. Graphical abstract A biosensor for vanillin was developed on the basis of an aptamer that was obtained via Capture-SELEX and by using an ISFET. This biosensor can be used for vanillin detection in presence of interferents and in real sample using an approach of ssDNA probe dehybridization.
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The electrochemical detection of interactions between aptamers and low-molecular-weight targets often lacks sensitivity. Signal amplification improves the detection of the aptamer-analyte complex; Bsm DNA polymerase was used to amplify the signal from the interaction of vanillin and its aptamer named Van_74 on an ion-sensitive field-effect transistor (ISFET)-based biosensor. The aptamer was immobilized on the ISFET sensitive surface. A short DNA probe was hybridized with the aptamer and dissociated from it upon vanillin addition. A free probe interacted with a special DNA molecular beacon initiated the Bsm DNA polymerase reaction that was detected by ISFET. A buffer solution suitable for both aptamer action and Bsm DNA polymerase activity was determined. The ISFET was shown to detect the Bsm DNA polymerase reaction under the selected conditions. Vanillin at different concentrations (1 × 10-6-1 × 10-8 M) was detected using the biosensor with signal amplification. The developed detection system allowed for the determination of vanillin, starting at a 10-8 M concentration. Application of the Bsm DNA polymerase resulted in a 15.5 times lower LoD when compared to the biosensor without signal amplification (10.1007/s00604-017-2586-4).
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Benzaldehídos/química , Aptámeros de Nucleótidos , Técnicas Biosensibles , ADN Polimerasa Dirigida por ADN , Técnicas Electroquímicas , OroRESUMEN
We demonstrate a proof-of-concept refractive index sensor based on heavily doped ZnO:Ga nanostructured in a grating configuration, which supports free space excitation of propagating surface plasmons. The bulk sensitivity of the sensor of 4.9 × 10(3) nm per refractive index unit, achieved in the mid-infrared spectral range with the first grating prototype, surpasses that of the noble metal counterparts by three to four times. Sensing performance is discussed in the light of numerical simulations of the spatial profile of the near field of surface plasmon polaritons.