Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 591
Filtrar
Más filtros

Medicinas Complementárias
Intervalo de año de publicación
1.
Sensors (Basel) ; 23(22)2023 Nov 15.
Artículo en Inglés | MEDLINE | ID: mdl-38005573

RESUMEN

Selenium (Se), as one of the essential and nutrient components of living organisms and plants, plays an important role in life activities, while excessive selenium is hazardous to human health. So, the establishment of an effective method for simple, rapid, and highly sensitive determination of selenium content is crucial in the field of food composition analysis and other areas. In this paper, a novel and simple ratiometric fluorescence method for the determination of Se has been developed using 9-anthracenemethanol (AM) as the ratiometric fluorescence reagent on the basis of the conventional fluorometric assay which utilized 2,3-diaminonapthalene (DAN) as fluorescent ligand. The ratiometric method was compared with the conventional method with respect to precision and accuracy. The inter-day and intra-day precisions (RSDs) of the ratiometric fluorescence method ranged from 2.08 to 2.78% and 1.28 to 1.84%, with mean recoveries of 93.2~98.0% and limit of detection (LOD) and limit of quantification (LOQ) of 0.0016 and 0.0049 µg/mL, respectively. This method was successfully applied to the determination of total selenium in selenium-enriched milk and selenium-supplemented shampoo, with the results in agreement with those obtained by inductively coupled plasma mass spectrometer (ICP-MS). The results demonstrated that the precision and accuracy of the ratiometric fluorescence method were superior to those of the conventional fluorescence method, and the interferences of various environmental factors were effectively eliminated. The precision and accuracy of the conventional method can be significantly improved by simply adding an elaborately selected ratiometric fluorescence reagent, and the new method will have broader practical applications.


Asunto(s)
Selenio , Humanos , Selenio/análisis , Espectrometría de Masas/métodos , Límite de Detección , Fluorometría , Colorantes
2.
Sci Rep ; 13(1): 12807, 2023 08 07.
Artículo en Inglés | MEDLINE | ID: mdl-37550465

RESUMEN

Labile copper(II) ions (Cu2+) in serum are considered to be readily available for cellular uptake and to constitute the biologically active Cu2+ species in the blood. It might also be suitable to reflect copper dyshomeostasis during diseases such as Wilson's disease (WD) or neurological disorders. So far, no direct quantification method has been described to determine this small Cu2+ subset. This study introduces a fluorometric high throughput assay using the novel Cu2+ binding fluoresceine-peptide sensor FP4 (Kd of the Cu2+-FP4-complex 0.38 pM) to determine labile Cu2+ in human and rat serum. Using 96 human serum samples, labile Cu2+was measured to be 0.14 ± 0.05 pM, showing no correlation with age or other serum trace elements. No sex-specific differences in labile Cu2+ concentrations were noted, in contrast to the total copper levels in serum. Analysis of the effect of drug therapy on labile Cu2+ in the sera of 19 patients with WD showed a significant decrease in labile Cu2+ following copper chelation therapy, suggesting that labile Cu2+ may be a specific marker of disease status and that the assay could be suitable for monitoring treatment progress.


Asunto(s)
Degeneración Hepatolenticular , Oligoelementos , Humanos , Ratas , Animales , Cobre/metabolismo , Degeneración Hepatolenticular/metabolismo , Fluorometría , Iones
3.
J Fluoresc ; 33(6): 2479-2488, 2023 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-37154848

RESUMEN

Manganese oxide nanoparticles (MnO Nps), sonochemical synthesized and characterized in our laboratory, are proposed as fluorescent sensor for selenium (Se) determination. The new methodology has been developed based on the enhancing effect of the Se(IV) on fluorescent emission of MnO Nps. Experimental variables that influence on fluorimetric sensitivity were optimized. The calibration graph using zeroth order regression was linear from 0.189 ng L-1 to 8.00 × 103 µg L-1, with correlation coefficient better than 0.99. Under the optimal conditions, the limits of detection and quantification were of 0.062 ng L-1 and 0.189 ng L-1, respectively. The trueness of the methodology was assessed through standard addition method obtaining recovery near to 100%. This method showed good tolerance to foreign ions, particularly to Se(VI), and was applied to determination of Se(IV) trace in food and drink samples with satisfactory results. With the intention of preserving the environment from harmful effects, a degradation study of the used nanomaterials has been included for their subsequent disposal.


Asunto(s)
Nanopartículas , Selenio , Selenio/química , Óxidos , Fluorometría , Colorantes
4.
Photochem Photobiol ; 99(3): 1010-1019, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-36094140

RESUMEN

Nonphotochemical quenching (NPQ) is known to depress in vivo fluorescence (IVF) of chlorophyll a (Chla) in aquatic environments, which makes it difficult to interpret the hour-to-hour variations in Chla measured by in situ fluorometers. We hypothesized that ratios between quenched and unquenched IVF are a function of both NPQ and photochemical quenching. In this study, two diatom model species Thalassiosira pseudonana (CCMP1335) and Thalassiosira weissflogii (CCMP1047) incubated under a sinusoidal light:dark cycle were studied; IVF was recorded continuously, and Chla and photo-physiological variables were measured seven times a day. The maximal decline in Chla-specific IVF (IVFB ) attributable to quenching was 50% under the experimental settings. An NPQ and photochemical quenching-based modeling equation exhibited a better match to the measured IVFB than equations representing the sole NPQ effect. Photochemical quenching induced by measuring light beam varied substantially during the day, and the part of the model for this process is excitation intensity-dependent (which is differed between models of in situ fluorometers, implying no straightforward method to correct Chla for all instrument models, instrument-specific parameterization is required). The forms of the IVFB -light relationship are discussed as well. The findings foster a holistic understanding of NPQ effects on in vivo Chla fluorometry.


Asunto(s)
Clorofila , Diatomeas , Clorofila A , Luz , Fluorometría/métodos , Fluorescencia , Complejo de Proteína del Fotosistema II
5.
Appl Microbiol Biotechnol ; 106(19-20): 6733-6743, 2022 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-36058939

RESUMEN

Rapid and accurate detection and identification of Staphylococcus aureus (S. aureus) are of great significance for food safety, environmental monitoring, early clinical diagnosis, and prevention of the spread of drug-resistant bacteria. Herein, we design a fluorometric aptasensor for ultra-sensitive, specific, and rapid detection of S. aureus. The apasensor combines the enrichment and separation of magnetic nanoparticles (MNPs), the biotin-streptavidin conjugation system, and a single S. aureus can release four signaling probes for signal amplification. Aptamer acts as a specific biorecognition element of S. aureus. Four FAM-labeled partially complementary sequences (FAM-pcDNAs) were used as signaling probes. The aptamers were sequential hybridized with the four FAM-pcDNAs to form aptamer&pcDNAs, which were then bound to MNPs via the biotin-streptavidin. When the aptamer specifically recognizes and binds to S. aureus, the FAM-pcDNAs signaling probes are replaced and released into the supernatant. The concentration of S. aureus can be quantified by measuring the fluorescence intensity (λexc/em = 492/520 nm) of the replaced signaling probe FAM-pcDNAs. The results show that the proposed fluorometric aptasensor displays good specificity, ultra-high sensitivity (1.23 cfu/mL), wide linear range (1 ~ 108 cfu/mL), and fast detection speed (~ 1.5 h). The recovery test verifies further that the proposed fluorometric aptasensor can detect S. aureus in spiked blood samples. Since aptamers are easy to customize, we believe that fluorometric aptasensors based on multiple amplification have broad prospects in the construction of practical high-performance biosensors for bacterial detection. KEY POINTS: • Multiple amplification-based fluorometric aptasensor for S. aureus is developed • The aptasensor displays high specificity with a LOD of 1.23 CFU/mL • The aptasensor can directly detect S. aureus in spiked blood samples.


Asunto(s)
Aptámeros de Nucleótidos , Técnicas Biosensibles , Infecciones Estafilocócicas , Técnicas Biosensibles/métodos , Biotina , Fluorometría/métodos , Humanos , Límite de Detección , Infecciones Estafilocócicas/diagnóstico , Staphylococcus aureus/genética , Estreptavidina
6.
Sensors (Basel) ; 22(5)2022 Mar 05.
Artículo en Inglés | MEDLINE | ID: mdl-35271186

RESUMEN

This study focuses on broadening the knowledge of a fluorometric index to improve the detection of oil substances present in the marine environment. It is assumed that the value of this index will provide information about a possible oil discharge at some distance from the sensor. In this paper, the detection of oil present in seawater as a mixture of oils such as fuel, lubricate oil, or crude oil based on a fluorescence indicator-fluorometric index (FIo/w) is discussed. FIo/w was defined based on specific excitation and emission wavelengths coming from the obtained excitation-emission spectrum (EEM) of oil-free seawater and, in parallel, the same water but artificially polluted with oil. For this, measurements of a mixture of oils in seawater for an oil-to-water ratio in the range from 50 × 10-9 to 200 × 10-9 as well as oil-free seawater were performed. Laboratory measurements continued five times in months in the summer season with the coastal waters of the southern Baltic Sea (last spring, summer, and early autumn). The dependence of FIo/w on the presence of oil in seawater, the oil-in-water ratio, as well as months of the considered season has been demonstrated.


Asunto(s)
Contaminación por Petróleo , Petróleo , Fluorometría , Aceites , Contaminación por Petróleo/análisis , Agua de Mar
7.
Talanta ; 242: 123305, 2022 May 15.
Artículo en Inglés | MEDLINE | ID: mdl-35183979

RESUMEN

Cadmium ion (Cd (II)) is a highly toxic heavy metal usually found in natural water. Exposure to Cd (II) can produce serious effects in human organs such as Itai-Itai disease. Therefore, the maximum allowance levels of Cd (II) in drinking water and herbal medicines imposed by the World Health Organization (WHO) are 3 µg L-1 and 300 µg kg-1, respectively. In this work, nitrogen-doped graphene quantum dots (N-GQDs) as a fluorescent sensor for Cd (II) determination was developed in both solution-based and paper-based systems. N-GQDs were synthesized from citric acid (CA) and ethylenediamine (EDA) via the hydrothermal method. The synthesized N-GQDs emitted intense blue fluorescence with a quantum yield (QY) of up to 80%. The functional groups on the surface of N-GQDs measured by FTIR were carboxyl (COO-), hydroxyl (OH-), and amine (NH2) groups, suggesting that they could be bound to Cd (II) for complexation. The fluorescence intensity of N-GQDs was gradually enhanced with the increase of Cd (II) concentration. This phenomenon was proved to result from the fluorescence enhancement (turn-on) based on the chelation enhanced fluorescence (CHEF) mechanism. Under the optimum conditions in the solution-based and paper-based systems, the limits of detection (LODs) were found to be 1.09 and 0.59 µg L-1, respectively. Furthermore, the developed sensors showed relatively high selectivity toward Cd (II) over ten other metal cations and six other anions of different charges. The performance of the sensor in real water and herbal medicine samples exhibited no significant difference as compared to the results of the validation method (ICP-OES). Therefore, the developed sensors can be used as fluorescent sensors for Cd (II) determination with high sensitivity, high selectivity, short incubation time (5 min). As such, the paper-based strategy has excellent promising potential for practical analysis of Cd (II) in water and herbal medicine samples with a trace level of Cd (II) concentrations.


Asunto(s)
Grafito , Puntos Cuánticos , Cadmio , Fluorometría , Humanos , Nitrógeno
8.
Anal Methods ; 13(32): 3551-3560, 2021 08 28.
Artículo en Inglés | MEDLINE | ID: mdl-34292282

RESUMEN

This work reports a facile synthesis of nitrogen-doped amorphous carbon nanodots (N-CNDs) and their use as a fluorometric paper-based sensor for the determination of Pb2+ at a low concentration. Both solution-based and paper-based systems were developed. The results show that the linearity ranges for Pb2+ determination were 0.010-10 mg L-1 (LOD = 0.008 mg L-1) and 0.005-0.075 mg L-1 (LOD = 0.004 mg L-1) for the solution-based and the paper-based sensors, respectively. Furthermore, the developed sensors show relatively high selectivity toward Pb2+ over ten other metal cations of different charges including As3+, Hg2+, Cd2+, Mg2+, Ni2+, Zn2+, Fe3+, Cu2+, Ba2+, and Ag+. The mechanism of Pb2+ determination was also investigated. It was found that the sensors exploited the quenching of the fluorescence intensity of N-CNDs by Pb2+via the photo-induced electron transfer (PET) mechanism. When applied to real water and herbal medicine samples, the performance of the sensor exhibited no significant difference as compared to the results of the validation method (ICP-OES). Overall, the developed sensors, especially the paper-based one, are promising for the practical analysis of Pb2+ in pharmaceutical and environmental samples with a low Pb2+ concentration.


Asunto(s)
Mercurio , Nitrógeno , Carbono , Fluorometría , Iones
9.
ACS Appl Mater Interfaces ; 13(30): 35365-35375, 2021 Aug 04.
Artículo en Inglés | MEDLINE | ID: mdl-34286953

RESUMEN

Phototherapy exhibits significant potential as a novel tumor treatment method, and the development of highly active photosensitizers and photothermal agents has drawn considerable attention. In this work, S and N atom co-doped carbon dots (S,N-CDs) with an absorption redshift effect were prepared by hydrothermal synthesis with lysine, o-phenylenediamine, and sulfuric acid as raw materials. The near-infrared (NIR) absorption features of the S,N-CDs resulted in two-photon (TP) emission, which has been used in TP fluorescence imaging of lysosomes and tumor tissue pH and real-time monitoring of apoptosis during tumor phototherapy, respectively. The obtained heteroatom co-doped CDs can be used not only as an NIR imaging probe but also as an effective photodynamic therapy/photothermal therapy (PDT/PTT) therapeutic agent. The efficiencies of different heteroatom-doped CDs in tumor treatment were compared. It was found that the S,N-CDs showed higher therapeutic efficiency than N-doped CDs, the efficiency of producing 1O2 was 27%, and the photothermal conversion efficiency reached 34.4%. The study provides new insight into the synthesis of carbon-based nanodrugs for synergistic phototherapy and accurate diagnosis of tumors.


Asunto(s)
Antineoplásicos/uso terapéutico , Colorantes Fluorescentes/uso terapéutico , Neoplasias/diagnóstico por imagen , Neoplasias/tratamiento farmacológico , Fármacos Fotosensibilizantes/uso terapéutico , Puntos Cuánticos/uso terapéutico , Animales , Antineoplásicos/química , Antineoplásicos/efectos de la radiación , Apoptosis/efectos de los fármacos , Carbono/química , Carbono/efectos de la radiación , Colorantes Fluorescentes/química , Colorantes Fluorescentes/efectos de la radiación , Fluorometría , Células HeLa , Humanos , Concentración de Iones de Hidrógeno , Lisosomas/metabolismo , Ratones Desnudos , Neoplasias/metabolismo , Nitrógeno/química , Nitrógeno/efectos de la radiación , Fotones , Fármacos Fotosensibilizantes/química , Fármacos Fotosensibilizantes/efectos de la radiación , Fototerapia , Puntos Cuánticos/química , Puntos Cuánticos/efectos de la radiación , Oxígeno Singlete/metabolismo , Azufre/química , Azufre/efectos de la radiación
10.
Int J Mol Sci ; 22(9)2021 Apr 29.
Artículo en Inglés | MEDLINE | ID: mdl-33947115

RESUMEN

Cortisol, a stress hormone, plays key roles in mediating stress and anti-inflammatory responses. As abnormal cortisol levels can induce various adverse effects, screening cortisol and cortisol analogues is important for monitoring stress levels and for identifying drug candidates. A novel cell-based sensing system was adopted for rapid screening of cortisol and its functional analogues under complex cellular regulation. We used glucocorticoid receptor (GR) fused to a split intein which reconstituted with the counterpart to trigger conditional protein splicing (CPS) in the presence of targets. CPS generates functional signal peptides which promptly translocate the fluorescent cargo. The sensor cells exhibited exceptional performance in discriminating between the functional and structural analogues of cortisol with improved sensitivity. Essential oil extracts with stress relief activity were screened using the sensor cells to identify GR effectors. The sensor cells responded to peppermint oil, and L-limonene and L-menthol were identified as potential GR effectors from the major components of peppermint oil. Further analysis indicated L-limonene as a selective GR agonist (SEGRA) which is a potential anti-inflammatory agent as it attenuates proinflammatory responses without causing notable adverse effects of GR agonists.


Asunto(s)
Técnicas Biosensibles , Evaluación Preclínica de Medicamentos/métodos , Polarización de Fluorescencia/métodos , Hidrocortisona/análisis , Aceites Volátiles/farmacología , Receptores de Glucocorticoides/agonistas , Atrofia , Acetato de Ciproterona/farmacología , Dexametasona/farmacología , Estradiol/farmacología , Fluorometría , Células HeLa , Humanos , Inteínas , Limoneno/farmacología , Proteínas Luminiscentes/análisis , Mentha piperita , Mentol/farmacología , Mifepristona/farmacología , Estructura Molecular , Músculo Esquelético/patología , Mioblastos/efectos de los fármacos , Aceites de Plantas/farmacología , Empalme de Proteína , Transporte de Proteínas , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/metabolismo , Proteína Fluorescente Roja
11.
Molecules ; 26(6)2021 Mar 12.
Artículo en Inglés | MEDLINE | ID: mdl-33809185

RESUMEN

INTRODUCTION: Alpha-galactosidase (α-Gal) is an enzyme responsible for the hydrolyzation of glycolipids and glycoprotein commonly found in dietary sources. More than 20% of the general population suffers from abdominal pain or discomfort caused by intestinal gas and by indigested or partially digested food residuals. Therefore, α-Gal is used in dietary supplements to reduce intestinal gases and help complex food digestion. Marketed enzyme-containing dietary supplements must be produced in accordance with the Food and Drug Administration (FDA) regulations for Current Good Manufacturing Practice (cGMPs). AIM: in this work we illustrated the process used to develop and validate a spectrophotometric enzymatic assay for α-Gal activity quantification in dietary supplements. METHODS: The validation workflow included an initial statistical-phase optimization of materials, reagents, and conditions, and subsequently a comparative study with another fluorimetric assay. A final validation of method performance in terms of specificity, linearity, accuracy, intermediate-precision repeatability, and system precision was then executed. RESULTS AND CONCLUSIONS: The proven method achieved good performance in the quantitative determination of α-Gal activity in commercial food supplements in accordance with the International Council for Harmonisation of Technical Requirements for Pharmaceuticals (ICH) guidelines and is suitable as a rapid in-house quality control test.


Asunto(s)
Suplementos Dietéticos/análisis , alfa-Galactosidasa/análisis , Suplementos Dietéticos/normas , Estabilidad de Enzimas , Fluorometría/métodos , Análisis de los Alimentos/métodos , Análisis de los Alimentos/normas , Análisis de los Alimentos/estadística & datos numéricos , Humanos , Laboratorios , Control de Calidad , Espectrofotometría/métodos , Estados Unidos , United States Food and Drug Administration , alfa-Galactosidasa/normas
12.
J Photochem Photobiol B ; 219: 112186, 2021 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-33892284

RESUMEN

Harmful algal blooms in inland waters are widely linked to excess phosphorus (P) loading, but increasing evidence shows that their growth and formation can also be influenced by nitrogen (N) and iron (Fe). Deficiency in N, P, and Fe differentially affects cellular photosystems and is manifested as changes in photosynthetic yield (Fv/Fm). While Fv/Fm has been increasingly used as a rapid and convenient in situ gauge of nutrient deficiency, there are few rigorous comparisons of instrument sensitivity and ability to resolve specific nutrient stresses. This study evaluated the application of Fv/Fm to cyanobacteria using controlled experiments on a single isolate and tested three hypotheses: i) single Fv/Fm measurements taken with different PAM fluorometers can distinguish among limitation by different nutrients, ii) measurements of Fv/Fm made by the addition of DCMU are comparable to PAM fluorometers, and iii) dark adaptation is not necessary for reliable Fv/Fm measurements. We compared Fv/Fm taken from the bloom-forming Microcystis aeruginosa (UTEX LB 3037) grown in nutrient-replete treatment (R) and N-, P-, and Fe-limited treatments (LN, LP, LFe, respectively), using three pulse-amplitude modulated (PAM) fluorometers and the chemical photosynthesis inhibitor 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU), and evaluated the effects of dark adaptation prior to PAM measurement. There were significant differences in Fv/Fm estimates among PAM fluorometers for light- versus dark-adapted cell suspensions over the whole experiment (21 days), which were all significantly higher than the DCMU-based measurements. However, dark adaptation had no effect on Fv/Fm when comparing PAM-based values across a single nutrient treatment. All Fv/Fm methods could distinguish LN and LP from R and LFe treatments but none were able to resolve LFe from R, or LN from LP cultures. These results indicated that for most PAM applications, dark adaptation is not necessary, and furthermore that single measurements of Fv/Fm do not provide a robust measurement of nutrient limitation in Microcystis aeruginosa UTEX LB 3037, and potentially other, common freshwater cyanobacteria.


Asunto(s)
Fluorometría/métodos , Microcystis/metabolismo , Nutrientes/química , Clorofila/química , Diurona/farmacología , Floraciones de Algas Nocivas/efectos de los fármacos , Floraciones de Algas Nocivas/efectos de la radiación , Hierro/química , Luz , Microcystis/crecimiento & desarrollo , Microcystis/efectos de la radiación , Nitrógeno/química , Nutrientes/farmacología , Fósforo/química , Fotosíntesis/efectos de los fármacos , Fotosíntesis/efectos de la radiación
13.
Biochem Biophys Res Commun ; 545: 125-131, 2021 03 19.
Artículo en Inglés | MEDLINE | ID: mdl-33548625

RESUMEN

Bromodomain and PHD finger containing transcription factor (BPTF) is a multidomain protein that regulates the transcription of chromatin and is related to many cancers. Herein, we report the screening-based discovery of Cpd1, a compound with micromolar affinity to the BPTF bromodomain. Through structure-guided optimization, we synthesized a variety of new inhibitors. Among these compounds, Cpd8 and Cpd10 were highly potent and selective inhibitors, with KD values of 428 nM and 655 nM in ITC assays, respectively. The high activity was explained by the cocrystal structure of Cpd8 in complex with the BPTF bromodomain protein. Cpd8 and Cpd10 were able to stabilize the BPTF bromodomain protein in cells in a cellular thermal shift assay (CETSA). Cpd8 downregulated c-MYC expression in A549 cells. All experiments prove that these two compounds are potential BPTF inhibitors.


Asunto(s)
Proteínas del Tejido Nervioso/antagonistas & inhibidores , Factores de Transcripción/antagonistas & inhibidores , Células A549 , Antígenos Nucleares/química , Antígenos Nucleares/genética , Calorimetría , Cristalografía por Rayos X , Diseño de Fármacos , Descubrimiento de Drogas , Evaluación Preclínica de Medicamentos , Fluorometría , Regulación de la Expresión Génica/efectos de los fármacos , Genes myc , Células HEK293 , Humanos , Modelos Moleculares , Proteínas del Tejido Nervioso/química , Proteínas del Tejido Nervioso/genética , Fragmentos de Péptidos/antagonistas & inhibidores , Fragmentos de Péptidos/química , Fragmentos de Péptidos/genética , Dominios Proteicos , Estabilidad Proteica/efectos de los fármacos , ARN Mensajero/genética , ARN Mensajero/metabolismo , Proteínas Recombinantes/química , Proteínas Recombinantes/efectos de los fármacos , Proteínas Recombinantes/genética , Relación Estructura-Actividad , Factores de Transcripción/química , Factores de Transcripción/genética
14.
Environ Geochem Health ; 43(2): 837-854, 2021 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-32328895

RESUMEN

The research is a maiden study aimed to assess the radioactivity in groundwater of Srinagar City using uranium and radon as proxies. In this study, 60 water samples were collected from various water sources that include bore wells, hand pumps and lakes of Srinagar City. Among them, 45 samples were taken from groundwater with depths ranging from 6 to - 126 m and the rest of the 15 samples were collected from surface sources like lakes, rivers and tap water. A gamma radiation survey of the area was carried out prior to collection of water samples, using a gamma radiation detector. A scintillation-based detector was utilized to measure radon, while as LED fluorimetry was employed to assess uranium in water samples. The average uranium concentration was found to be 2.63 µg L-1 with a maximum value of 15.28 µg L-1 which is less than the globally accepted permissible level of 30 µg L-1. 222Radon concentration varied from 0.2 to 38.5 Bq L-1 with an average value of 8.9 Bq L-1. The radon concentration in 19 groundwater samples (32% of total sites) exceeded the permissible limits of 11 Bq L-1 set by USEPA. This information could be of vital importance to health professionals in Kashmir who are researching on the incidence of lung cancers in the region given the fact that radon is the second leading cause of lung cancers after smoking worldwide.


Asunto(s)
Carcinógenos Ambientales/análisis , Agua Subterránea/química , Monitoreo de Radiación/métodos , Contaminantes Radiactivos del Agua/análisis , Ciudades , Fluorometría , Humanos , Radón/análisis , Conteo por Cintilación , Uranio/análisis
15.
Biometals ; 33(4-5): 255-267, 2020 10.
Artículo en Inglés | MEDLINE | ID: mdl-32979113

RESUMEN

The nematode Caenorhabditis elegans (C. elegans) is a convenient tool to evaluate iron metabolism as it shares great orthology with human proteins involved in iron transport, in addition to being transparent and readily available. In this work, we describe how wild-type (N2) C. elegans nematodes in the first larval stage can be loaded with acetomethoxycalcein (CAL-AM) and study it as a whole-organism model for both iron speciation and chelator permeability of the labile iron pool (LIP). This model may be relevant for high throughput assessment of molecules intended for chelation therapy of iron overload diseases.


Asunto(s)
Fluorometría , Quelantes del Hierro/química , Animales , Caenorhabditis elegans , Quelantes del Hierro/síntesis química , Estructura Molecular
16.
Org Biomol Chem ; 18(22): 4157-4163, 2020 06 10.
Artículo en Inglés | MEDLINE | ID: mdl-32458889

RESUMEN

There are relatively few methods available for discovering inhibitors of the protein-protein interactions (PPIs) that hold together homo-oligomers. We envisioned that Differential Scanning Fluorimetry (DSF) might be a versatile way to discover this type of inhibitor because oligomers are often more thermally stable than monomers. Using the homo-heptameric chaperonin, Hsp60, as a model, we screened ∼5000 diverse compounds in 384-well plates by DSF, revealing molecules that partially inhibited oligomerization. Because DSF does not require protein labeling or structural information, we propose that it could be a versatile way to uncover PPI inhibitors.


Asunto(s)
Chaperonina 60/antagonistas & inhibidores , Fluorometría , Relación Dosis-Respuesta a Droga , Evaluación Preclínica de Medicamentos , Estructura Molecular , Unión Proteica/efectos de los fármacos
17.
Anal Chim Acta ; 1101: 120-128, 2020 Mar 08.
Artículo en Inglés | MEDLINE | ID: mdl-32029102

RESUMEN

Simple and easy to engineer metal-sensing molecules that are capable of differentiating metal ions and producing metal-specific signals are highly desirable. Metal ions affect the thermal stability of proteins by increasing or decreasing their resistance to unfolding. This work illustrates a new strategy for designing bivalent fluorescent fusion proteins capable of differentiating metal ions in solution through their distinct effects on a protein's thermal stability. A new dual purpose metal sensor was developed consisting of biotin protein ligase (BirA) from B. pseudomallei (Bp) fused to green fluorescent protein (GFP). When coupled with differential scanning fluorimetry of GFP-tagged proteins (DSF-GTP) for signal-transduction detection, Bp BirA-GFP yields distinct protein unfolding signatures with Zn(II) and Cu(II) ions in aqueous solutions. The limit of detection of the system is ∼1 µM for both metal species. The system can be used in a variety of high-throughput assay formats including for the screening of metal-binding proteins and chelators. Bp BirA-GFP has also the additional benefit of being useful in Cu(II) ion field-testing applications through simple visual observation of a temperature-dependent loss of fluorescence. Bp BirA-GFP is the first example of a 2protein-based dual purpose Cu(II) and Zn(II) ion sensor compatible with two different yet complementary signal-transduction detection systems.


Asunto(s)
Ligasas de Carbono-Nitrógeno/química , Cobre/análisis , Proteínas Fluorescentes Verdes/química , Proteínas Recombinantes de Fusión/química , Zinc/análisis , Proteínas Bacterianas/química , Proteínas Bacterianas/metabolismo , Técnicas Biosensibles/métodos , Biotina/metabolismo , Burkholderia pseudomallei/enzimología , Ligasas de Carbono-Nitrógeno/metabolismo , Cobre/metabolismo , Fluorometría/métodos , Proteínas Fluorescentes Verdes/metabolismo , Límite de Detección , Prueba de Estudio Conceptual , Unión Proteica , Proteínas Recombinantes de Fusión/metabolismo , Zinc/metabolismo
18.
Mikrochim Acta ; 187(3): 158, 2020 02 07.
Artículo en Inglés | MEDLINE | ID: mdl-32034503

RESUMEN

D-penicillamine and histidine-functionalized graphene quantum dot (DPA-GQD-His) was synthesized and applied in a fluorometric method for determination of acetamiprid using a G-quadruplex DNAzyme. At first DNA probe (probe 1) consists of a target-specific aptamer with two arms of DNA segments. Probe 1 was hybridized with DNA probe 2 composed of a single DNA sequence with two split G-rich DNA sequences. This leads to the formation of a triplex-to-G-quadruplex (TPGQ). Next, acetamiprid was hybridized with the aptamer in the TPGQ to release free DNA probe 2. The released probe 2, in the presence of of K+, undergoes a structural change into a stem-loop structure (by self-complementary hybridization and Hoogsteen hydrogen bonding) that bears a G-quadruplex structure. This is followed by conjugation with hemin to form the G-quadruplex/hemin DNAzyme. The DNAzyme catalyzes the oxidation of o-phenylenediamine by H2O2 to produce a yellow fluorescent product with excitation/emission maxima at 420/560 nm. The oxidation product interacts with DPA-GQD-His to achieve a rapid energy transfer between DPA-GQD-His and oxidation product. This increases the fluorescence of the oxidation product and quenches the fluorescence of DPA-GQD-His. DPA-GQD-His also improves the catalytic activity of DNAzyme towards oxidation of ophenylenediamine oxidization and enhances fluorometric response to acetamiprid. The assay works in the 1.0 fM to 1.0 nM acetamiprid concentration range and has a 0.38 fM detection limit. It was successfully applied to the determination of acetamiprid in tea. Graphical abstractThe study reported one double amplification strategy for ultrasensitive fluorescence detection of acetamiprid in tea with D-penicillamine and histidine-functionalized graphene quantum dots and G-quadruplex/heminDNAzyme. The analtyical method exhibits ultra high sensitivity, selectivity and rapidity of fluorescence response to acetamiprid.


Asunto(s)
Aptámeros de Nucleótidos/química , Técnicas Biosensibles/métodos , Fluorometría/métodos , Grafito/química , Hemina/química , Neonicotinoides/química , Puntos Cuánticos/química , Histidina , Humanos , Penicilamina
19.
Mikrochim Acta ; 187(1): 81, 2020 01 02.
Artículo en Inglés | MEDLINE | ID: mdl-31897750

RESUMEN

"Tryptophan-coated blue fluorescent copper nanocluster (CuNC@Trp) was prepared by a strategy where Trp acts as both the reducing and capping agent. The fluorescence of the CuNC, with excitation/emission peaks at 340/405 nm, is selectively quenched by iron(II) and iron(III) ions. Studying the mechanism of this interaction revealed that Fe2+ and Fe3+ ions can make a ground state complex with the protecting ligand which can result in quenching of the cluster emission. Structural and optical properties of the modified CuNC were investigated by ESI-MS, DLS, TEM, UV-vis and photoluminescence. The effects of pH value and temperature, time of interaction, and cluster volume were optimized. Under optimized conditions, the probe response is linear in concentration range of 10-1000 µM for Fe(II) and Fe(III) with the relative standard deviations of 0.13 and 0.14% (n = 5) respectively. The respective limits of detection are 3.0 and 2.2 µM. The method was successfully used for determination of trace amount of both ions in spiked water, blood and iron supplement tablets. The results were in good agreement with those obtained by the ICP-AES method." Graphical abstractThe scheme represents the synthesis of CuNC@Trp at basic conditions and at elevated temperature. The emission of the cluster decreases due to static quenching of fluorescence by iron ions.


Asunto(s)
Cobre/química , Fluorescencia , Fluorometría/métodos , Hierro/análisis , Nanopartículas del Metal/química , Triptófano/química , Fluorometría/normas , Concentración de Iones de Hidrógeno , Iones/análisis , Iones/química , Hierro/química , Análisis Espectral , Temperatura
20.
Mikrochim Acta ; 187(1): 85, 2020 01 02.
Artículo en Inglés | MEDLINE | ID: mdl-31897844

RESUMEN

A fluorescent nanoprobe for Pb(II) has been developed by employing aptamer-functionalized upconversion nanoparticles (UCNPs) and magnetic Fe3O4-modified (MNPs) gold nanoparticles (GNPs). First, aptamer-functionalized UCNPs and aptamer-functionalized magnetic GNPs were synthesized to obtained the fluorescent nanoprobe. The particles were combined by adding a complementary ssDNA. In the absence of Pb(II), the UCNPs, MNPs and GNPs are linked via complementary base pairing. This led to a decrease in the green upconversion fluorescence peaking at 547 nm (under 980 nm excitation). In the presence of Pb(II), the dsDNA between UCNPs and MNPs-GNPs is cleaved, and fluorescence recovers. This effect allows Pb(II) to be quantified, with a wide working range of 25-1400 nM and a lower detection limit of 5.7 nM. The nanoprobe gave satisfactory results when analyzing Pb(II) in tea and waste water. Graphical abstractSchematic representation of fluorescent nanoprobe based on fluorescence resonance energy transfer (FRET) between upconversion nanoparticles (UCNPs) and gold nanoparticles (GNPs)-Fe3O4 magnetic nanoparticles (MNPs) for detection of Pb2+.


Asunto(s)
Aptámeros de Nucleótidos , Óxido Ferrosoférrico/química , Fluorometría/métodos , Oro , Plomo/análisis , Nanopartículas del Metal/química , Nanopartículas/química , Emparejamiento Base , ADN de Cadena Simple/química , Transferencia Resonante de Energía de Fluorescencia/métodos , Fluorometría/normas , Té/química , Aguas Residuales/química
SELECCIÓN DE REFERENCIAS
DETALLE DE LA BÚSQUEDA