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1.
J Fluoresc ; 33(4): 1651-1659, 2023 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-36806048

RESUMO

In this study, with the help of reactive monomers, crosslinkers, and photoinitiator that detect H2S in various matrices, an H2S sensitive fluorescence sensor polymerizes under ultraviolet (UV) light was developed. To this goal, a polymeric membrane was prepared, and the characterization of the membrane was carried out with Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM) methods. Afterward, appropriate conditions were identified, the excitation wavelength was determined as 370 nm, and the emission wavelength was determined as 425 nm. It was established that the fluorescence intensity of the prepared polymeric membrane decreased in the presence of H2S. A detailed analysis was executed to determine the sensor's most suitable pH value and time. It was found that the optimum pH was 8.0, and the optimal duration was 15 s. It has been calculated that the linear range of the developed method is 2.19 × 10-8- 6.25 × 10-7 M, and the detection limit (LOD) is 7.37 × 10-9 M. The effect of some possible interfering ions was investigated, and it determined that the sensor had excellent selectivity. In addition, the sensor used to determine H2S can be used at least 100 times. The recovery percentages were 102.1%-103.2%, and 104.6%, using tap water samples. In terms of providing reliable, fast results, high sensitivity, reusable, low cost, and ease of use, the developed fluorimetric sensor, compared to standard methods, has become more advantageous.

2.
Protein Expr Purif ; 202: 106187, 2023 02.
Artigo em Inglês | MEDLINE | ID: mdl-36216219

RESUMO

Recombinant expression and purification of proteins have become a staple of modern drug discovery as it enables more precise in vitro analyses of drug targets, which may help obtain biochemical and biophysical parameters of a known enzyme and even uncover unknown characteristics indicative of novel enzymatic functions. Such information is often necessary to prepare adequate screening assays and drug-discovery experiments in general. Toxoplasma gondii is an obligate protozoan parasite that is a member of the phylum Apicomplexa, can develop several neuro-degenerative symptoms and, in specific cases, certain death for human hosts. Its relict non-photosynthetic plastid, the apicoplast, harbours a unique de novo long-chain fatty acid synthesis pathway of a prokaryotic character, FASII. The FASII pathway shows plasticity and, is essential for many intracellular and membranal components, along with fatty acid uptake via salvaging from the host, therefore, its disruption causes parasite death. TgFabG, a FASII enzyme responsible for a single reduction step in the pathway, was recombinantly expressed, purified and biochemically and biophysically characterised in this study. The bioengineering hurdle of expressing the recombinant gene of a eukaryotic, signal peptide-containing protein in a prokaryotic system was overcome for the apicomplexan enzyme TgFabG, by truncating the N-terminal signal peptide. TgFabG was ultimately recombinantly produced in a plasmid expression vector from its 1131 base pair gene, purified as 260 and 272 amino acid proteins using a hexahistidine (6 × Histag) affinity chromatography and its biochemical (enzyme activity and kinetics) and biophysical characteristics were analysed in vitro.


Assuntos
Apicoplastos , Toxoplasma , Humanos , Apicoplastos/metabolismo , Toxoplasma/genética , Toxoplasma/metabolismo , Proteína de Transporte de Acila/metabolismo , Oxirredutases/metabolismo , Ácidos Graxos/metabolismo , Sinais Direcionadores de Proteínas , Proteínas de Protozoários/genética , Proteínas de Protozoários/metabolismo
3.
Food Chem ; 394: 133525, 2022 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-35749880

RESUMO

Melamine selective acrylate citric acid (ACA) based polymeric membrane sensor was prepared by radical polymerization method and the sensor was characterized. The sensor showed a selective fluorescent response to melamine (λex/λem = 388/425 nm). The sensor response is linear in the concentration range of 3.96 × 10-9 to 7.93 × 10-8 mol L-1, the optimum pH value is 6.0 and response time is less than 1 min. Limit of detection (LOD) and limit of quantification (LOQ) were calculated as 2.32 × 10-10 mol L-1 and as 7.74 × 10-10 mol L-1, respectively. The sensor showed great selectivity towards melamine in the presence of a large number of molecules and ions. The performance of sensor was also confirmed by determining of melamine in milk powder sample and the results were compared with HPLC results and acceptable results were obtained. As a conclusion, the results revealed that the proposed sensor is an interesting alternative for melamine determination.


Assuntos
Contaminação de Alimentos , Leite , Animais , Ácido Cítrico/análise , Contaminação de Alimentos/análise , Limite de Detecção , Leite/química , Polímeros/análise , Pós/análise , Triazinas/análise
4.
Anal Sci ; 36(7): 807-811, 2020 Jul 10.
Artigo em Inglês | MEDLINE | ID: mdl-31904005

RESUMO

A novel polymeric membrane sensor was developed by using 2-hydroxyethyl acrylate, trimethylolpropane triacrylate, (3-mercaptopropyl)trimethoxysilane, and poly(ethylene glycol)diacrylate for As(III) determination. Various parameters, like the pH, response time, the foreign ions, and concentration effects were investigated for deciding the optimum working conditions of the polymeric sensor. As a result of this investigation, the optimum pH was found to be 2, and the response time was found to be 30 s. The linear range of the sensor was 6.65 × 10-9 - 3.99 × 10-8 mol L-1 (0.50 - 2.99 µg L-1) with a detection limit of 2.33 × 10-9 mol L-1 (0.18 µg L-1). Soy flour and well-water samples were successfully analyzed with the developed sensor. The sensor can be used at least 100 times after regeneration. It could be reused by washing with purified water and showed good stability for 6 months.

5.
Sci Total Environ ; 601-602: 405-424, 2017 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-28570975

RESUMO

This work was implemented to study the physicochemical and biological characteristics of the Sea of Marmara and the Turkish straits (TSS: Bosporus and Dardanelles straits) for the period 2010-2013 and to calculate winter and summer fluxes of nutrients (nitrates, phosphates) between the Aegean and Black Seas through the TSS. The brackish Black Sea waters reach the Dardanelles Strait with modified biochemical properties. The salinity and phosphates of the surface waters increased westwards. Biologically labile nutrients of Black Sea origin are utilized through biological processes in the Marmara Sea. On the other hand, increase of nutrients due to land based sources has led to eutrophication problems in the area. The sub surface water layer of Mediterranean origin is oxygen depleted (saturation<30%) and rich in nutrients. Higher oxygen values indicated water mixing of the Sea of Marmara during winter 2012. Ammonium was the predominant form of inorganic nitrogen. The study area has been classified from Moderate to Bad trophic status. Izmit Bay also faced serious eutrophication problems together with hypoxic conditions below the halocline. Nutrient fluxes through the TSS showed temporal variation in the upper and lower layers related to changes in both nutrient concentrations and the water mass volume fluxes. Surface nitrates and phosphates outflux from the Sea of Marmara to the Aegean Sea was higher than the influx from the Black Sea through Bosporus strait, indicating high enrichment of nutrients in the Sea of Marmara from anthropogenic sources.


Assuntos
Monitoramento Ambiental , Eutrofização , Poluentes da Água/análise , Mar Negro , Nitratos/análise , Nitrogênio/análise , Fosfatos/análise , Salinidade , Turquia
6.
Spectrochim Acta A Mol Biomol Spectrosc ; 159: 106-12, 2016 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-26836450

RESUMO

In this study, 1-vinylimidazole units bearing photocured films were prepared as fluorescent sensors towards Pb(2+) in aqueous solutions. The influence of experimental parameters such as pH, time and foreign ion concentrations were investigated. Sensor response was linear over a concentration range of 4.83×10(-8) to 4.83×10(-7) mol L(-1). The sensor was highly sensitive with a detection limit as low as 1.87×10(-8)molL(-1), and having a selectivity of over four thousand fold. The response time of the sensor was found to be 5 min. When stored in a desiccator at room temperature the sensor showed good stability after a 5 month period. The fluorescence sensors were successful in the determination of Pb(2+) in water samples as well as in the determination of the quantitative amount of lead and the results were satisfying. Compared with previously reported literature, the prepared new sensor is highly sensitive and selective.

7.
Anal Chim Acta ; 812: 215-21, 2014 Feb 17.
Artigo em Inglês | MEDLINE | ID: mdl-24491784

RESUMO

This study describes the preparation and the characterization of a new thiol-ene based polymeric fluorescence sensor by photo initiated polymerization of trimethylolpropane tris(3-mercaptopropionate), 2-hydroxyethylacrylate, and 2,4,6-triallyloxy-1,3,5-triazine which are used as monomers and also a photo initiator (2,2-dimethoxy-2-phenylacetophenone) for its usage as optical sensor for gold ions. The thiol-ene based polymeric membrane sensor was characterized by using attenuated total reflectance-fourier transform infrared spectroscopy (ATR-FTIR) and scanning electron microscopy (SEM). The response characteristics of the sensors including dynamic range, pH effect, response time, and the effect of foreign ions were investigated. Fluorescence spectra showed that the excitation/emission maxima of the membrane were at 379/425 nm, respectively.

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