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1.
Vet Med Sci ; 10(4): e1491, 2024 Jul.
Article in English | MEDLINE | ID: mdl-39031626

ABSTRACT

BACKGROUND: Haemagglutinin-neuraminidase (HN) is one of the membrane proteins of Newcastle disease virus (NDV) that plays a significant role during host viral infection. Therefore, antibodies against HN are vital for the host's ability to protect itself against NDV infection due to their critical functions in viral infection. As a result, HN has been a candidate protein in vaccine development against the Newcastle disease virus. METHODS: This report used the full-length sequence of the HN protein of NDV isolated in Iran (VIId subgenotype). We characterize and identify amino acid substitutions in comparison to other more prevalent NDV genotypes, VII subgenotypes and vaccine strains. Furthermore, bioinformatics tools were applied to determine the three-dimensional structure, molecular dynamics simulation and prediction of B-cell antigenic epitopes. RESULTS: The results showed that the antigenic regions of our isolate are quite comparable to the other VII subgenotypes of NDV isolated from different geographical places. Moreover, by employing the final 3D structure of our HN protein, the amino acid residues are proposed as a B-cell epitope by epitope prediction servers, which leads to the introduction of linear and conformational antigenic sites. CONCLUSIONS: Immunoinformatic vaccine design principles currently exhibit tremendous potential for developing a new generation of candidate vaccines quickly and economically to eradicate infectious viruses, including the NDV. In order to accomplish this, focus is directed on residues that might be considered antigenic.


Subject(s)
Genotype , HN Protein , Newcastle disease virus , Newcastle disease virus/genetics , Newcastle disease virus/immunology , HN Protein/genetics , HN Protein/chemistry , Amino Acid Sequence , Animals , Iran , Base Sequence , Chickens , Poultry Diseases/virology , Newcastle Disease/virology
2.
Lab Chip ; 24(8): 2298-2305, 2024 Apr 16.
Article in English | MEDLINE | ID: mdl-38517043

ABSTRACT

A volumetric bar-chart chip (V-chip) is a microfluidic device based on distance-based quantitative measurement that visualizes analyte concentration without the need for apparatus or data processing. This typically utilizes special receptors and catalysis parts that generate oxygen, so ink can be moved inside the channels, and enables instant visual quantitation of the analyte. However, the low stability of some macromolecules, the use of expensive catalysts, and difficulty in controlling the process cause inaccurate readings, and therefore, limit further development and the use of these systems. In this article, we introduced a novel approach that eliminates the use of catalysts in V-chips and provides an efficient and simple path in the design of biosensors. The product of the enzymatic reaction of urease with urea is bicarbonate, which turns into CO2 gas in an acidic environment. Therefore, the amount of gas produced is proportional to the amount of urea in the sample, and it can be quantitatively measured by visual detection from the amount of ink movement caused by CO2 gas pressure. This biosensor has a linear response range of 0 to 1000 µg ml-1 and a detection limit of 3.6 µg ml-1 in raw milk. The recovery of urea in raw milk at 100 and 400 µg ml-1 concentrations was 96.5% and 98.9%, respectively. This volumetric chip shows potential for determining urea levels in real samples without requiring additional equipment. The combination of the sensitivity and specificity of enzymatic reactions, inherent gas-generating reactions, and the processability of microchips discussed in this paper can be the basis for the comprehensive development of volumetric chips, which can create a new path for the development of efficient and cheap biosensors.


Subject(s)
Aptamers, Nucleotide , Biosensing Techniques , Carbon Dioxide , Lab-On-A-Chip Devices , Equipment Design , Urea
3.
Comput Biol Med ; 163: 107087, 2023 09.
Article in English | MEDLINE | ID: mdl-37321098

ABSTRACT

Infectious Bursal Disease (IBD) is a common and contagious viral infection that significantly affects the poultry industry. This severely suppresses the immune system in chickens, thereby threating their health and well-being. Vaccination is the most effective strategy for preventing and controlling this infectious agent. The development of VP2-based DNA vaccines combined with biological adjuvants has recently received considerable attention due to their effectiveness in eliciting both humoral and cellular immune responses. In this study, we applied bioinformatics tools to design a fused bioadjuvant candidate vaccine from the full-length sequence of the VP2 protein of IBDV isolated in Iran using the antigenic epitope of chicken IL-2 (chiIL-2). Furthermore, to improve the antigenic epitope presentation and to maintain the three-dimensional structure of the chimeric gene construct, the P2A linker (L) was used to fuse the two fragments. Our in-silico analysis for the design of a candidate vaccine indicates that a continuous sequence of amino acid residues ranging from 105 to 129 in chiIL-2 is proposed as a B cell epitope by epitope prediction servers. The final 3D structure of the VP2-L-chiIL-2105-129 was subjected to physicochemical property determination, molecular dynamic simulation, and antigenic site determination. The results of these analyses led to the development of a stable candidate vaccine that is non-allergenic and has the potential for antigenic surface display potential and adjuvant activity. Finally, it is necessary to investigate the immune response induced by our proposed vaccine in avian hosts. Notably, increasing the immunogenicity of DNA vaccines can be achieved by combining antigenic proteins with molecular adjuvants using the principle of rational vaccine design.


Subject(s)
Infectious bursal disease virus , Vaccines, DNA , Animals , Interleukin-2/genetics , Infectious bursal disease virus/genetics , Chickens , Vaccines, DNA/genetics , Epitopes , Antibodies, Viral , Adjuvants, Immunologic/genetics
4.
J Struct Biol ; 215(1): 107925, 2023 03.
Article in English | MEDLINE | ID: mdl-36470559

ABSTRACT

Staphylococcal protein A (SpA) domain B (the basis of affibody) has been widely used in affinity chromatography and found therapeutic applications against inflammatory diseases through targeting the Fc part of immunoglobulin G (IgG). We have performed extensive molecular dynamics simulation of 41 SpA mutants and compared their dynamics and conformations to wild type. The simulations revealed the molecular details of structural and dynamics changes that occurred due to introducing point mutations and helped to explain the SPR results. It was observed in some variants a point mutation caused extensive structural changes far from the mutation site, while an effect of some other mutations was limited to the site of the mutated residue. Also, the pattern of hydrogen bond networks and hydrophobic core arrangements were investigated. We figured out mutations that occurred at positions 128, 136, 150 and 153, affected two hydrophobic cores at the interface as well as mutations introduced at positions 129 and 154 interrupted two hydrogen bond networks of the interface, SPR data showed all of these mutations reduced binding affinity significantly. Overall, by scanning the SpA-Fc interface through the large numbers of introduced mutations, the new insights have been gained which would help to design high- affinity ligands of IgG.


Subject(s)
Immunoglobulin G , Molecular Dynamics Simulation , Immunoglobulin G/genetics , Immunoglobulin G/chemistry , Immunoglobulin G/metabolism , Mutation/genetics , Protein Binding/genetics , Immunoglobulin Fc Fragments/immunology
5.
Phys Chem Chem Phys ; 24(35): 21178-21187, 2022 Sep 14.
Article in English | MEDLINE | ID: mdl-36039911

ABSTRACT

We present a new water-dependent molecular mechanism for the widely-used protein stabilizing osmolyte, trimethylamine N-oxide (TMAO), whose mode of action has remained controversial. Classical interpretations, such as osmolyte exclusion from the vicinity of protein, cannot adequately explain the behavior of this osmolyte and were challenged by recent data showing the direct interactions of TMAO with proteins, mainly via hydrophobic binding. Solvent effect theories also fail to propose a straightforward mechanism. To explore the role of water and the hydrophobic association, we disabled osmolyte-protein hydrophobic interactions by replacing water with hexane and using lipase enzyme as an anhydrous-stable protein. Biocatalysis experiments showed that under this non-aqueous condition, TMAO does not act as a stabilizer, but strongly deactivates the enzyme. Molecular dynamics (MD) simulations reveal that TMAO accumulates near the enzyme and makes many hydrogen bonds with it, like denaturing osmolytes. Some TMAO molecules even reach the active site and interact strongly with the catalystic traid. In aqueous solvent, the enzyme functions well: the extent of TMAO interactions is reduced and can be divided into both polar and non-polar terms. Structural analysis shows that in water, some TMAO molecules bind to the enzyme surface like a surfactant. We show that these interactions limit water-protein hydrogen bonds and unfavorable water-hydrophobic surface contacts. Moreover, a more hydrophobic environment is formed in the solvation layer, which reduces water dynamics and subsequently, rigidifies the backbone in aqueous solution. We show that osmolyte amphiphilicity and protein surface heterogeneity can address the weaknesses of exclusion and solvent effect theories about the TMAO mechanism.


Subject(s)
Methylamines , Proteins , Hydrophobic and Hydrophilic Interactions , Methylamines/chemistry , Proteins/chemistry , Solvents/chemistry , Urea/chemistry , Water/chemistry
6.
Chemosphere ; 304: 135305, 2022 Oct.
Article in English | MEDLINE | ID: mdl-35718034

ABSTRACT

Among the important needs of human societies is the elimination of environmental pollution and also the construction of high-performance and inexpensive biosensors. In this regard, the construction of multi-functional composites has been considered. A novel magnetic graphite carbon nitride decorated by tri-vanadium substituted Dawson-type heteropolytungstate nanocomposite (C3N4/Fe3O4@P2W15V3) effectively synthesized and characterized by prevalent functional analysis. The prepared nano-catalyst presents bi-functional usage involving photocatalytic removal of dyes (methylene blue, congo red and phenyl red) (around 98%) under visible light radiation and greatly sensitive colorimetric sensing of cysteine in an aqueous media. Moreover, synthesized nano-catalyst successfully recovered five times without any considerable deficiency on its photocatalytic ability. Further, Moreover, we propose a novel method for cysteine detection based on the C3N4/Fe3O4@P2W15V3 nanocomposite. This nanocomposite displayed a privileged catalytic feature for cysteine oxidation to extend a clock reaction of methylene blue as an indicator in the presence of NaBH4 in acidic solution. More importantly, this colorimetric sensing method of cysteine presents an easy, low-cost, selective, and rapid colorimetric assay with a detection limit value of 7.2 µM in the acceptable linear range of 5-600 µM.


Subject(s)
Cysteine , Nanocomposites , Catalysis , Coloring Agents , Humans , Light , Methylene Blue
7.
Comput Biol Med ; 140: 105057, 2021 Nov 24.
Article in English | MEDLINE | ID: mdl-34839187

ABSTRACT

Vaccination remains the most effective strategy for preventing and controlling infectious diseases. Numerous conventional vaccines, especially live attenuated, inactivated (killed) microorganisms and subunit vaccines, lead to an effective induction of protective immune responses, mainly antibody-mediated responses against pathogens. However, it has become known that a wide range of highly dangerous pathogens are uncontrollable via conventional vaccination strategies. Recent advances in molecular biology, immunology, genetics, biochemistry, and bioinformatics have provided new prospects for vaccine development. As a result of these advances, several new strategies for vaccine design, development, and production have appeared. These strategies show advantages over conventional vaccines. In this review, we discuss some of the major novel approaches, including recombinant protein vaccines, live recombinant viral and bacterial vectors, DNA and RNA vaccines, reverse vaccinology and reverse genetics approaches. Moreover, we have described the recent progresses on computational tools and immunoinformatics approaches for identifying, designing, and developing new candidate vaccines.

8.
Int J Biol Macromol ; 170: 189-195, 2021 Feb 15.
Article in English | MEDLINE | ID: mdl-33333096

ABSTRACT

The activity of Horseradish Peroxidase (HRP) Enzyme exposed to a static magnetic field (SMF) during the oxidation reaction of pyrogallol (PGL) and the epigallocatechin gallate (EPCG) flavonoid was recorded at different times. As the data showed, the enzyme activity increased by 77.17% with increasing incubation time up to 30 min. The kinetic parameters KM and Vmax for PGL sample incubated in SMF for 30 min were 5.641 × 10-3 mM, 4.424 × 10-2 mmol/min, respectively, and for EPCG sample with the same condition were 8.65 × 10-4 mM, 2.37 × 10-3 mmol/min, respectively. Exposure of HRP enzyme to SMF changed the optimum pH from 7.0 to 6.0 in 10 min, but did not create any change in the optimum temperature of the enzyme. After 120 h, the residual activity of normal enzyme was 17% higher than that of the incubated enzyme. The structural changes of the control and HRP enzyme incubated in SMF were investigated by relative viscosity, fluorescence and CD, UV-Vis spectrophotometry. The structural changes in the presence of SMF were found to cause changes in the enzyme activity. In fact, changes in the amount of hydrogen bonds between enzymes and solvents can be a reason for this behavior from a molecular point of view. Using a static magnetic field can provide a new approach to control and direct enzyme-based biological processes.


Subject(s)
Horseradish Peroxidase/chemistry , Magnetic Fields , Catechin/analogs & derivatives , Catechin/metabolism , Circular Dichroism , Horseradish Peroxidase/metabolism , Hydrogen-Ion Concentration , Kinetics , Molecular Structure , Oxidation-Reduction , Pyrogallol/metabolism , Solvents , Spectrometry, Fluorescence , Spectrophotometry, Ultraviolet , Structure-Activity Relationship , Temperature , Viscosity
9.
J Biomol Struct Dyn ; 39(9): 3071-3079, 2021 Jun.
Article in English | MEDLINE | ID: mdl-32323612

ABSTRACT

This study introduces a computational method to design a new aptamer with higher binding affinity to a special target in comparison with the experimentally available aptamers. The method is called step by step mutation based on MD simulation, which includes some steps. First, MD simulation is performed for the SELEX-introduced (native) aptamer in the presence of the target. Afterwards, conformational factor (Pi) is calculated for the simulated system, which obtains the affinity of the aptamer residues to the target. A nucleotide exchange is done for the residue with the least Pi parameter to the nucleotide with the highest Pi value that results in a mutant aptamer. MD simulation is performed for the target-mutant complex, and Pi values are calculated again. The nucleotide exchange is performed similarly, and the designing process is proceeded repeatedly that results in a mutant with the improved specificity to the target. The aptamer affinity to the target is also determined in each step through calculating the binding Gibbs energy (ΔGBind) as a reliable parameter. The introduced strategy is utilized efficiently to design a mutant aptamer with improved specificity toward sulfadimethoxine (SDM) antibiotic as a case study. The great difference in the ΔGBind values about 579.856 kJ mol-1 highlights that the M5 mutant possesses the improved specificity toward SDM in comparison with the native aptamer. Besides, the selectivity of the M5 aptamer toward SDM is examined among some conventional interfering compounds by using MD simulation that confirms the applicability of the designed aptamer for further experimental studies.


Subject(s)
Aptamers, Nucleotide , Sulfadimethoxine , Anti-Bacterial Agents , Computers , Mutation
10.
J Mol Graph Model ; 101: 107733, 2020 12.
Article in English | MEDLINE | ID: mdl-32931983

ABSTRACT

Aggregation behavior of dodecyl betaine chloride [DB][Cl], as an amino acid ionic liquid, and dodecyl betaine N-acetyl glycinate [DB][AG], as a bio ionic liquid, in aqueous media was studied through molecular dynamics simulation. The aggregating was investigated by radial distribution function, coordination numbers, and hydrogen bond numbers. The results demonstrated the hydrogen bond between [DB]+ and [AG]- that leads to aggregation. The number of hydrogen bonds of [DB][AG] is greater than [DB][Cl] and causes a decrease in the gradient of the mean square displacement, thereby the diffusion coefficient of cation, anion, and water in [DB][AG]. The results point to a stable aggregation of [DB][AG] which is in agreement with the results of root mean square deviations. The aggregation number for [DB][AG] is 50 and 44 for [DB][Cl]. Computing the radius of gyration and geometrical radius shows that the aggregation size is 23.0 Å and 26.4 Å for [DB][AG] and [DB][Cl], respectively. It was also observed that the shape of the aggregates is quasi-spherical that points to a sub-diffusive regime.


Subject(s)
Ionic Liquids , Amino Acids , Anions , Hydrogen Bonding , Molecular Dynamics Simulation
11.
Biotechnol Prog ; 36(6): e3035, 2020 11.
Article in English | MEDLINE | ID: mdl-32519466

ABSTRACT

The present study aimed to investigate, for the first time, the rate of the oxidation reaction of some derivatives of phenol and aromatic amines, that is, pyrogallol, catechol, resorcinol, ortho-aminophenol, meta-aminophenol, para-aminophenol, ortho-phenylenediamine, and para-phenylenediamine, in the presence of hydrogen peroxide in pure and magnetized solvents using horseradish peroxidase enzyme. The reaction was studied in the absence and presence of a magnetized solvent under completely identical conditions. The results showed that magnetized solvent could change the structure of the enzyme and reduce its activity. In addition, it affected the rate of oxidation of the selected derivatives through altering the strength of the hydrogen bonds of the system. The changes in the structure and activity of the enzyme were examined using UV-Vis and fluorescence spectroscopy as well as viscosity measurement technique. Examination of the secondary structure via the far UV-CD spectrum indicated the increase in the alpha helical structure in the magnetized solvent. When dissolved in a magnetized solvent, hydrogen peroxide as an enzyme substrate reduced the rate of enzymatic reaction and provided lower saturation conditions for the enzyme compared with when it was dissolved in the pure solvent.


Subject(s)
Amines/pharmacology , Horseradish Peroxidase/chemistry , Phenols/chemistry , Water/chemistry , Amines/chemistry , Magnetic Phenomena , Oxidation-Reduction/drug effects , Oxidation-Reduction/radiation effects , Phenols/pharmacology , Ultraviolet Rays , Water/pharmacology
12.
Food Sci Anim Resour ; 40(3): 350-361, 2020 Apr.
Article in English | MEDLINE | ID: mdl-32426715

ABSTRACT

Quality control of meat products is one of the main concerns of consumers, governmental control authorities, and retailers. The purpose of this study was to employ ribose-induced Maillard reaction in detection of meat adulteration and differentiation of fresh-chilled from frozen-thawed minced veal. The browning intensity was assessed through measuring the absorbance at 420 nm with a spectrophotometer as well as the direct analysis of the color and pH. The results showed that CIE b*, CIE a*, and A420* values in the extract of fresh-chilled veal were significantly (p<0.05) higher than frozen-thawed samples. The extract of frozen meat samples stored at -18°C became significantly darker and more yellowish compared to -4°C. The results showed that the A420* value in the frozen-thawed veal stored at -4°C and -18°C was reduced by approximately 17.22±3.53% and 11.68±2.49%, respectively, compared with fresh-chilled veal. The findings also showed that the storage temperature of minced veal and the heating time in this reaction had a significant effect on all tested variables (p<0.0001). The proposed method can be considered as an easy, quick, and inexpensive test for differentiating between the fresh-chilled and frozen-thawed minced veal.

13.
Comput Biol Chem ; 86: 107267, 2020 Jun.
Article in English | MEDLINE | ID: mdl-32470911

ABSTRACT

In this work, systematic density functional theory (DFT) calculations were performed to study the interactions of various metal ions (Al3+, Fe3+, Co2+, Ni2+, Cu2+, and Zn2+) and the clinically useful chelating agent called deferiprone (DFP) at the M05-2X/6-31G(d) level of theory. The thermodynamic parameters of metal-deferiprone complexes were determined in water. Based on the obtained data, the theoretical binding energy trend is as follows: Al3+ > Fe3+ > Cu2+ > Ni2+ > Co2+ > Zn2+, confirming that [Al(DFP)3] has the most interaction energy. Moreover, Natural bond orbital analysis was employed to determine and analyze the natural charges on different atoms and charge transfer between the metal ions and ligands (oxygen atoms) as well as the interaction energy (E(2)) values. The calculated value of Æ©E(2) (donor-acceptor interaction energy) for [Al(DFP)3] complex is higher than other complexes, which is according to energy analysis. To confirm the type of effective interactions and bonding properties in the water, the quantum theory of atoms in molecules (QTAIM) analysis was applied. QTAIM analysis confirmed that the strongest M - O bond is found in the [Al(DFP)3] complex. The calculated topological properties at the bond critical points, such as the ratio of the kinetic energy density to the potential energy density, -G(r)/V(r), electronic energy density, H(r), confirm that M - O bonds in the Al-deferiprone complex are non-covalent, while in other complexes, they are electrostatic and partially covalent.

14.
Anal Biochem ; 597: 113689, 2020 05 15.
Article in English | MEDLINE | ID: mdl-32199832

ABSTRACT

Mercury (Hg2+) and silver (Ag+) ions possess the harmful effects on public health and environment that makes it essential to develop the sensing techniques with great sensitivity for the ions. Metal ions commonly coexist in the different biological and environmental systems. Hence, it is an urgent demand to design a simple method for the simultaneous detection of metal ions, peculiarly in the case of coexisting Hg2+ and Ag+. This study introduces a low-cost paper-based aptasensor to monitor Hg2+ and Ag+, simultaneously. The strategy of the sensing array is according to the conformational changes of Hg2+- and Ag+-specific aptamers and their release from the GO surface after the injection of the target sample on the sensing platform. Through monitoring the fluorescence recovery changes against the concentrations of the ions, Hg2+ and Ag+ can be determined as low as 1.33 and 1.01 pM. The paper-based aptasensor can simultaneously detect the ions within about 10 min. The aptasensor is applied prosperously to monitor Hg2+ and Ag+ in human serum, water, and milk. The designed aptasensor with the main advantages of simplicity and feasibility holds the supreme potential to develop a cost-effective sensing method for environmental monitoring, food control, and human diagnostics.


Subject(s)
Aptamers, Nucleotide/chemistry , Biosensing Techniques , Mercury/analysis , Paper , Silver/analysis , Aptamers, Nucleotide/economics , Biosensing Techniques/economics , Graphite/chemistry , Graphite/economics , Mercury/economics , Silver/economics
15.
J Mol Graph Model ; 97: 107570, 2020 06.
Article in English | MEDLINE | ID: mdl-32097885

ABSTRACT

Chloroperoxidase (CPO) is a versatile fungal heme-thiolate protein that catalyzes a variety of one electron and two-electron oxidations. Chloroperoxidase is a versatile fungal heme-thiolate protein that catalyzes a variety of oxidations. CPO enzyme contains thirteen sugars, including five N-acetyl D-glucosamines (NAG) and eight mannoses (MAN), which are attached to the protein via the glycosidic bonds. Removal of the sugars from CPO leads to increase the hydrophobicity of the enzyme, as well as the reduction of the alkylation reactions. However, due to the lack of the proper force field for the sugars, they are ignored in the theoretical studies. The present study aims to assess the effects of the sugar segments on the structure and activity of CPO through the simulation of the halo structure and the structures without the sugar segment. Despite the difficulty of the process and being time-consuming, the suitable force field is introduced successfully for the sugars. According to molecular dynamics simulation (MD), seven channels and fifteen cavities are identified in the CPO structure. Two of the channels provide the substrate access to the active site. The MD simulation results reveal that the removal of NAG decreases the number of the cavities from fifteen to eleven. Besides, the removal of NAG is associated with removing the channel providing the substrate access. The number of the cavities decreases from fifteen to fourteen through the removal of MAN; however, channel providing the substrate access to the active site is partly preserved. The MD simulation results indicate that the structures without the sugar units are more compact in comparison with the halo structures. The removal of the sugar segments induces the significant changes in the flexibility of the residues that affect the catalytic activity of the enzyme. As a result, the enzyme activities, such as the oxidation, alkylation, halogenation, and epoxidation cannot occur when the sugar segments of the enzyme are removed.


Subject(s)
Chloride Peroxidase , Fungi/enzymology , Catalysis , Chloride Peroxidase/metabolism , Heme/metabolism , Molecular Dynamics Simulation , Oxidation-Reduction
16.
Phytochem Anal ; 31(5): 564-574, 2020 Sep.
Article in English | MEDLINE | ID: mdl-31997502

ABSTRACT

INTRODUCTION: Saffron (Crocus sativus L.) is a well-known spice which is used as the colourant and flavouring agent in food products. Safranal could act as an indicator for saffron grading, authentication and adulteration, as well as for quality evaluation of saffron flavoured products; since it is the main odourant and the most aroma-active compound of saffron. OBJECTIVES: Firstly, determination of the optimum static conditions for safranal extraction through headspace solid-phase micro-extraction combined with gas chromatography (HS-SPME-GC) technique. Secondly, safranal measurement in different saffron flavoured products under the optimised static conditions. Thirdly, elucidation of the method efficiency for safranal measurement under non-equilibrium conditions for a saffron drink sample. METHODS: Different equilibrium times, pH and salt concentrations were applied on aqueous solutions of safranal. Accordingly, the optimised static conditions were determined for safranal extraction through HS-SPME-GC approach using polydimethylsiloxane (PDMS) fibre. RESULTS: Under static conditions, a linear response was obtained for standard curve within the safranal concentration range of 0.08-30 ppm, with R2 = 0.9999. The limits of detection and quantification were 0.04 and 0.08 ppm, respectively. Despite the fact that safranal peak area was an efficient parameter for quantifications under static conditions; its poor reproducibility was proved under dynamic conditions for the saffron drink sample. This observation necessitated application of kinetic studies on real food samples. CONCLUSIONS: Safranal extraction was successfully performed from aqueous matrices through HS-SPME-GC, under static conditions. Mathematical modelling resulted in kinetic parameters that improved the efficiency of safranal measurement under dynamic conditions, using PDMS fibre.


Subject(s)
Benzenesulfonates , Chromatography, Gas , Cyclohexenes , Gas Chromatography-Mass Spectrometry , Kinetics , Reproducibility of Results , Terpenes
17.
J Biomol Struct Dyn ; 38(12): 3659-3675, 2020 Aug.
Article in English | MEDLINE | ID: mdl-31496379

ABSTRACT

The aptamers with the ability to form a G-quadruplex structure can be stable in the presence of some ions. Hence, study of the interactions between such aptamers and ions can be beneficial to determine the highest selective aptamer toward an ion. In this article, molecular dynamics (MD) simulations and quantum mechanics (QM) calculations have been applied to investigate the selectivity of the T30695 aptamer toward Pb2+ in comparison with some ions. The Free Energy Landscape (FEL) analysis indicates that Pb2+ has remained inside the aptamer during the MD simulation, while the other ions have left it. The Molecular Mechanics Poisson-Boltzmann Surface Area (MM-PBSA) binding energies prove that the conformational stability of the aptamer is the highest in the presence of Pb2+. According to the compaction parameters, the greatest compressed ion-aptamer complex, and hence, the highest ion-aptamer interaction have been induced in the presence of Pb2+. The contact maps clarify the closer contacts between the nucleotides of the aptamer in the presence of Pb2+. The density functional theory (DFT) results show that Pb2+ forms the most stable complex with the aptamer, which is consistent with the MD results. The QM calculations reveal that the N-H bonds and the O…H distances are the longest and the shortest, respectively, in the presence of Pb2+. The obtained results verify that the strongest hydrogen bonds (HBs), and hence, the most compressed aptamer structure are induced by Pb2+. Besides, atoms in molecules (AIM) and natural bond orbital (NBO) analyses confirm the results.Communicated by Ramaswamy H. Sarma.


Subject(s)
G-Quadruplexes , Molecular Dynamics Simulation , Density Functional Theory , Ions , Lead , Quantum Theory
18.
Life Sci ; 234: 116758, 2019 Oct 01.
Article in English | MEDLINE | ID: mdl-31421083

ABSTRACT

In this work, fluorescent copper oxide nanoparticles (CuO NPs) were green synthesized using viable cells, cell lysate supernatant (CLS) and protein extracts of luminescent Vibrio sp. VLC. Biogenic CuO NPs were then characterized by XRD, FTIR, UV/Vis spectroscopy, TEM, DLS, and PL spectroscopy. Results showed that CLS method was more efficient for CuO NPs production, therefore CuO NPs synthesized by this method from copper sulfate (CuO NPs-1) and/or copper nitrate (CuO NPs-2) were used for further studies. The crystallite size of polydispersed CuO NPs-1 and CuO NPs-2 were about 8.83 and 8.77 nm, respectively indicating their suitability for biological applications. Antibacterial activity of CuO NPs was determined using broth microdilution, well diffusion agar, and time-kill curves methods. Both CuO NP-1 and CuO NP-2 inhibited bacterial growth at the minimum inhibitory concentration (MIC) of 625 mg/L except St. mutants (MIC = 1250 mg/L). Emission of fluorescent light from the surface of NPs was increased when exposed to Cd2+, As2+ and Hg2+ ions but decreased by Pb2+ ions. Results showed that CuO NP-1 had anticancer properties against KYSE30 esophageal cancer cell line (IC50 = 13.96 mg/L) while no higher cytotoxic effects were observed on Human Dermal Fibroblasts (HDF) (IC50 = 48.88 mg/L).


Subject(s)
Anti-Bacterial Agents/pharmacology , Antineoplastic Agents/pharmacology , Copper/pharmacology , Esophageal Neoplasms/drug therapy , Fluorescent Dyes/chemistry , Metals, Heavy/analysis , Nanoparticles/chemistry , Anti-Bacterial Agents/chemistry , Antineoplastic Agents/chemistry , Bacteria/drug effects , Bacterial Infections/drug therapy , Cell Line, Tumor , Copper/chemistry , Green Chemistry Technology/methods , Humans , Spectrometry, Fluorescence/methods , Vibrio/chemistry
19.
IET Nanobiotechnol ; 13(2): 189-192, 2019 Apr.
Article in English | MEDLINE | ID: mdl-31051450

ABSTRACT

Silver nanoparticles (AgNPs) have been biosynthesised through the extracts of Ribes khorassanicum fruits, which served as the reducing agents and capping agents. Biosynthesised AgNPs have been found to be ultraviolet-visible (UV-vis) absorption spectra since they have displayed one surface plasmon resonance peak at 438 nm, attesting the formation of spherical NPs. These particles have been characterised by UV-vis, field-emission scanning electron microscopy, energy-dispersive X-ray spectroscopy, X-ray diffraction, Fourier transform infrared spectroscopy, and transmission electron microscopy analysis. The formation of AgNPs at 1.0 mM concentration of AgNO3 has resulted in NPs that contained mean diameters in a range of 20-40 nm. The green-synthesised AgNPs have demonstrated high antibacterial effect against pathogenic bacteria (i.e. Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa). Biosynthesising metal NPs through plant extracts can serve as the facile and eco-friendly alternative for chemical and/or physical methods that are utilised for large-scale nanometal fabrication in various medical and industrial applications.


Subject(s)
Anti-Bacterial Agents/chemistry , Metal Nanoparticles/chemistry , Plant Extracts/chemistry , Ribes/chemistry , Silver/chemistry , Anti-Bacterial Agents/metabolism , Anti-Bacterial Agents/pharmacology , Bacteria/drug effects , Microscopy, Electron, Transmission , Plant Extracts/metabolism , Silver/metabolism , Silver/pharmacology , Spectroscopy, Fourier Transform Infrared
20.
Anal Chim Acta ; 1071: 70-77, 2019 Sep 13.
Article in English | MEDLINE | ID: mdl-31128757

ABSTRACT

In this study, a simple paper-based aptasensor has been developed for the ultrasensitive detection of lead (Pb2+) ion within about 10 min. The aptasensor has been successfully designed by taking advantages of the Förster Resonance Energy Transfer (FRET) process and the super fluorescence quenching property of graphene oxide (GO) sheet. The sensing mechanism of the aptasensor is based on the conformational switch of the Pb2+-specific aptamer from a random coil to a G-quadruplex structure. An injection of Pb2+ on the paper-based platform induces the release of the specific aptamer from the GO surface that recovers the fluorescence emission. Under the optimal experimental conditions, there is a good linear relationship between the fluorescence recovery and the Pb2+concentration in the ranges of 5-70 pM and 0.07-20 nM. Moreover, the aptasensing array exhibits a high sensitivity to Pb2+ with an ultra-low detection limit of 0.5 pM. The developed aptasensor has been successfully applied to determine Pb2+ in tap water, lake water, milk, and human blood serum. The paper-based aptasensor can be efficiently utilized to detect other metal ions and biological molecules by substituting target specific aptamer.


Subject(s)
Aptamers, Nucleotide/chemistry , DNA/chemistry , Lead/blood , Paper , Animals , Biosensing Techniques/instrumentation , Biosensing Techniques/methods , Drinking Water/analysis , Fluorescence , Fluorescence Resonance Energy Transfer/instrumentation , Fluorescence Resonance Energy Transfer/methods , Food Contamination/analysis , G-Quadruplexes/drug effects , Graphite/chemistry , Lakes/analysis , Limit of Detection , Milk/chemistry
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